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Cited by 84Endometriosis Research and TreatmentNeuropeptides and Animal PhysiologyIon Channels and Receptors

Neuropeptides Substance P and Calcitonin Gene Related Peptide Accelerate the Development and Fibrogenesis of Endometriosis

Open access
Yan, Dingmin, Liu, Xishi, Guo, Sun-Wei·2019·Scientific Reports
DOI 10.1038/s41598-019-39170-wPMC PMC6389969
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Abstract

Endometriotic lesions are known to be hyperinnervated, especially in lesions of deep endometriosis (DE), which are frequently in close proximity to various nerve plexuses. DE lesions typically have higher fibromuscular content than that of ovarian endometriomas (OE) lesions, but the underlying reason remains elusive. Aside from their traditional role of pain transduction, however, whether or not sensory nerves play any role in the development of endometriosis is unclear. Here, we show that, thorough their respective receptors neurokinin receptor 1 (NK1R), calcitonin receptor like receptor (CRLR), and receptor activity modifying protein 1 (RAMP-1), neuropeptides substance P (SP) and calcitonin gene related peptide (CGRP) induce epithelial-mesenchymal transition (EMT), fibroblast-to-myofibroblast transdifferentiation (FMT) and further turn stromal cells into smooth muscle cells (SMCs) in endometriotic lesions, resulting ultimately in fibrosis. We show that SP and CGRP, or the rat dorsal root ganglia (DRG) supernatant, through the induction of NK1R and CGRP/CRLR/RAMP-1 signaling pathways, promoted EMT, FMT and SMM in endometriosis, resulting in increased migratory and invasive propensity, cell contractility, production of collagen, and eventually to fibrosis. Neutralization of NK1R and/or CGRP/CRLR/RAMP-1 abrogated these processes. Extended exposure of endometriotic stromal cells to SP and/or CGRP or the DRG supernatant induced increased expression of α-SMA, desmin, oxytocin receptor, and smooth muscle myosin heavy-chain. Finally, we show that DE lesions had significantly higher nerve fiber density, increased staining levels of α-SMA, NK1R, CRLR, and RAMP-1, concomitant with higher lesional fibrotic content than that of OE lesions. The extent of lesional fibrosis correlated positively with the staining levels of NK1R, CRLR, and RAMP-1, as well as the nerve fiber density in lesions. Thus, this study provides another piece of evidence that sensory nerves play an important role in promoting the development and fibrogenesis of endometriosis. It explains as why DE frequently have higher fibromuscular content than that of OE, highlights the importance of lesional microenvironment in shaping the lesional fate, gives more credence to the idea that ectopic endometrium is fundamentally wounds that go through repeated tissue injury and repair, and should shed much needed light into the pathophysiology of endometriosis.

AI snapshot

Written from the paper’s full text.

TL;DR

SP and CGRP, acting through NK1R and CGRP/CRLR/RAMP-1 signaling, promote EMT, FMT, and smooth muscle metaplasia in endometriotic and endometrial cells, increasing contractility and collagen production; receptor neutralization blocks these effects. In human lesions, deep endometriosis had higher nerve fiber density, receptor staining, and fibrosis than ovarian endometriomas, and lesional fibrosis correlated with NK1R/RAMP-1/CRLR expression and nerve fiber density.

Key findings

  • In 11Z endometriotic epithelial cells, SP and/or CGRP treatment for 72 h decreased E-cadherin and increased Snai1, Slug, vimentin, N-cadherin, and PAI-1 (all p < 0.05), and increased proliferation, migration, and invasion (migration all p < 0.001; proliferation and invasion all p < 0.05).
  • In ESCs and HESCs, SP and/or CGRP for 12 days upregulated CCN2, FN, LOX, COL1A1, and α-SMA (all p < 0.05), and increased α-SMA, desmin, OTR, and SM-MHC; contractility increased by 3 h and collagen production increased after 72 h (all p < 0.05).
  • DRG supernatant induced myofibroblast and SMC markers in HESCs, while aprepitant and/or CGRP fragment 8–37 largely abolished DRG-induced EMT, FMT, SMM, contractility, and collagen production.
  • In human tissues (30 OE, 30 DE, 24 normal endometrium), DE lesions had higher nerve fiber density, fibrosis, α-SMA, and NK1R than OE; RAMP-1 and CRLR were higher in OE and DE than normal endometrium. Lesional nerve fiber density correlated with dysmenorrhea severity (Spearman’s r = 0.85, p < 2.2 × 10−16), and fibrosis correlated with NK1R, RAMP-1, and CRLR staining (all r’s ≥ 0.60, p < 5.2 × 10−17).

Methods

  • In vitro experiments used the 11Z endometriotic epithelial cell line, an endometrial stromal cell line (ESCs), and primary human endometriotic stromal cells (HESCs) derived from ovarian endometriomas; treatments included SP and CGRP at 10−7 M, aprepitant at 10−6 M, CGRP fragment 8–37 at 10−6 M, and rat DRG supernatant.
  • Assays included CCK-8 proliferation, scratch migration, Matrigel invasion, real-time RT-PCR, Western blot, immunofluorescence, collagen gel contraction, and Sircol soluble collagen assays.
  • Human tissue analysis used immunohistochemistry for α-SMA, NK1R, CRLR, RAMP-1, and CGRP, plus Masson trichrome staining for fibrosis; DRG neurons were isolated from 30 virgin female Sprague-Dawley rats (4–5 weeks old, 100–120 g). Statistical analysis used R version 3.5.0 with Wilcoxon, paired t-test or paired Wilcoxon, Pearson, and Spearman tests.

Limitations

  • Stated: the study used an endometriotic epithelial cell line instead of primary endometriotic epithelial cells due to technical difficulty, and used ESCs plus HESCs derived from ovarian endometriomas, not from deep endometriosis lesions; these cells may differ phenotypically and functionally from DE-derived cells.
  • Stated: deriving epithelial cells from DE lesions may be impractical because of technical difficulty and because 12–15% of DE patients have “stromal endometriosis” with absent glandular epithelium, and highly fibromuscular DE lesions may contain few genuine endometriotic stromal cells.
  • Evident: the human lesion data are cross-sectional correlations, so they cannot establish causality between nerve fiber density, receptor expression, and fibrosis; the in vitro work relies heavily on cell lines and OE-derived primary cells rather than DE-derived cells.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.1038/s41598-019-39170-w

Cites

135 of 135 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

0 of 115 cited sources checked · 0 supported · 0 flagged

  • Findingown data

    Immunofluorescence confirmed progressively increased expression of α-SMA, F-actin, desmin, OTR and SM-MHC in HESCs treated with SP as treatment duration increased.

    “immunofluorescence confirmed the progressively increased expression of α-SMA, F-actin, desmin, OTR and SM-MHC in HESCs treated with SP as the treatment duration increased (Fig. 3C)”
    Quote in the paperSubstantial claim
  • Findingown data

    Endometriotic epithelial cells initially did not show any expression of α-SMA or F-actin.

    “Not surprisingly, endometriotic epithelial cells initially did not show any expression of α-SMA or F-actin (Fig. 2).”
    Quote in the paperSubstantial claim
  • Findingown data

    11Z cells treated with SP and/or CGRP for 72 hours resulted in significantly increased cellular proliferation compared with controls (all p<0.05).

    “we found that 11Z cells treated with SP and/or CGRP for 72 hours resulted in significantly increased cellular proliferation as compared with controls (all p-values < 0.05; Fig. 1D).”
    Quote in the paperSubstantial claim
  • Findingown data

    In women with endometriosis, the severity of dysmenorrhea correlated positively with the extent of fibrosis in endometriotic lesions (r = 0.90, p < 2.2 × 10−16).

    “It also positively correlated with the extent of fibrosis in endometriotic lesions (r = 0.90, p < 2.2 × 10−16; Supplementary Fig. 1B)”
    Quote in the paperNot a substantial claim
  • Findingown data

    Migratory capability of 11Z cells treated with SP and/or CGRP was significantly increased compared with untreated ones (all p<0.001).

    “the migratory capability of 11Z cells treated with SP and/or CGRP was significantly increased as compared with untreated ones (all p-values < 0.001; Fig. 1E).”
    Quote in the paperSubstantial claim
  • May combine two claims. 39
  • The quote only partly states this claim. 2
  • Classified as probably not a substantial claim. 27

  • May combine two claims.

    Findingown data

    Immunoreactivity against α-SMA was seen mostly in cytoplasm in endometriotic stromal cells, and NK1R staining was seen mostly in cytoplasm and membranes in endometriotic epithelial cells.

    “We found that the immunoreactivity against α-SMA was seen mostly in cytoplasm in endometriotic stromal cells, as expected, and NK1R staining was seen mostly in cytoplasm and membranes in endometriotic epithelial cells (Fig. 8A).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Compared with OE lesions, DE lesions have significantly higher nerve fiber density, higher extent of fibrosis, and higher α-SMA and NK1R staining levels.

    “we found that, compared with OE lesions, DE lesions have significantly higher nerve fiber density, higher extent of fibrosis, and higher α-SMA and NK1R staining levels (Fig. 8).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Contractility of 11Z cells, HESCs, and ESCs was all significantly increased after treatments with SP and/or CGRP (all p<0.05) just for 3 hours and further increased progressively and then plateaued at 48 hours.

    “the contractility of 11Z cells, HESCs, and ESCs was all significantly increased after treatments with SP and/or CGRP (all p-values < 0.05) just for 3 hours and further increased progressively and then plateaued at 48 hours”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    HESCs treated with DRG supernatant for 4 days had slightly increased α-SMA and F-actin staining but negative staining of OTR, desmin, and SM-MHC.

    “we found that HESCs treated with the DRG supernatant for 4 days had slightly increased α-SMA and F-actin staining but, not surprisingly, negative staining of OTR, desmin, and SM-MHC.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Cells treated with vehicle for the same durations were negative for vimentin, α-SMA and F-actin but positive for E-cadherin.

    “In contrast, cells treated with vehicle for the same durations were all negative for vimentin, α-SMA and F-actin but positive for E-cadherin as shown in Fig. 2A”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Neutralization of SP and/or CGRP abolished increased cellular contractility induced by DRG supernatant in endometriotic stromal cells, and abolished production of soluble collagens induced by DRG supernatant in endometriotic epithelial and stromal cells as well as endometrial stromal cells.

    “neutralization of SP and/or CGRP also abolished increased cellular contractility induced by the DRG supernatant in endometriotic stromal cells, and abolished the production of soluble collagens induced by the treatment with the DRG supernatant in endometriotic epithelial and stromal cells as well as endometrial stromal cells (Fig. 7C–D).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    DE lesions had significantly higher nerve fiber density, increased staining levels of α-SMA, NK1R, CRLR, and RAMP-1, concomitant with higher lesional fibrotic content than OE lesions.

    “Finally, we show that DE lesions had significantly higher nerve fiber density, increased staining levels of α-SMA, NK1R, CRLR, and RAMP-1, concomitant with higher lesional fibrotic content than that of OE lesions.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    RAMP-1 and CRLR immunostaining could be found in both epithelial and stromal components of normal endometrium, but their staining levels were significantly lower than that in either OE or DE lesions.

    “The RAMP-1 and CRLR immunostaining could be found in both epithelial and stromal components of normal endometrium, but their staining levels were significantly lower than that in either OE or DE lesions (Fig. 8A,B).”
    Quote in the paperSubstantial claim

  • The quote only partly states this claim.

    Findingown data

    Antagonism of NK1R, RAMP-1 and CRLR stalls SP/CGRP-induced EMT, FMT, SMC transdifferentiation and fibrogenesis.

    “Antagonism of their respective receptors, however, stalls these processes.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    DE is characterized by the presence of smooth muscle metaplasia (SMM) and a high degree of fibrotic tissues.

    “One feature that DE stands out from other subtypes of endometriosis is its presence of smooth muscle metaplasia (SMM) and its high degree of fibrotic tissues4,15–17”
    Quote in the paperSubstantial claim0 of 4 sources checked
    • cites [4] Cornillie, F. J., Oosterlynck, D., Lauweryns, J. M. & Koninckx, P. R. Deeply infiltrating pelvic endometriosis: histology and clinical significance. Fertil Steril 53, 978–983, S0015-0282(16)53570-5 (1990). source

      Not checked in the free run.

    • cites [15] Anaf, V., Simon, P., Fayt, I. & Noel, J. Smooth muscles are frequent components of endometriotic lesions. Hum Reprod 15, 767–771 (2000). source

      Not checked in the free run.

    • cites [16] Donnez, J. et al. Current thinking on the pathogenesis of endometriosis. Gynecol Obstet Invest 54 Suppl 1, 52–58; discussion 59–62, 66295 (2002). source

      Not checked in the free run.

    • cites [17] van Kaam, K. J., Schouten, J. P., Nap, A. W., Dunselman, G. A. & Groothuis, P. G. Fibromuscular differentiation in deeply infiltrating endometriosis is a reaction of resident fibroblasts to the presence of ectopic endometrium. Hum Reprod 23, 2692–2700, den153 (2008). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    As a result of ReTIAR, ectopic endometrium interacts with its microenvironment, activates TGF-β1/Smad3 signaling, experiences EMT and FMT, causing increased collagen production and cellular contractility and eventually fibrosis.

    “As a result of this ReTIAR, the ectopic endometrium actively interacts with various cells in its microenvironment, activates the transforming growth factor (TGF)-β1/Smad3 signaling and experiences epithelial-mesenchymal transition (EMT) and fibroblast-to-myofibroblast transdifferentiation (FMT), causing increased collagen production and cellular contractility and eventually resulting in fibrosis22”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [22] Zhang, Q., Duan, J., Liu, X. & Guo, S. W. Platelets drive smooth muscle metaplasia and fibrogenesis in endometriosis through epithelial-mesenchymal transition and fibroblast-to-myofibroblast transdifferentiation. Mol Cell Endocrinol 428, 1–16, S0303-7207(16)30063-6 (2016). source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    Sensory nerves induce endometriotic stromal cells to undergo further differentiation into fully differentiated SMCs and are thus responsible for SMM in endometriotic lesions.

    “indicating that sensory nerves induce endometriotic stromal cells to undergo further differentiation into fully differentiated SMCs and are thus responsible for SMM in endometriotic lesions.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: SP and CGRP are secreted mostly from sensory nerves.

  • May combine two claims.

    Premisecitation

    Compared with OE, DE lesions appeared to have gone through EMT, FMT, and SMM more thoroughly and extensively, and exhibited significantly more fibromuscular tissues yet reduced cellularity and vascularity.

    “we recently found that, compared with OE, DE lesions appeared to have gone through EMT, FMT, and SMM more thoroughly and more extensively, and, accordingly, exhibited significantly more fibromuscular tissues yet reduced cellularity and vascularity28.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [28] Liu, X., Zhang, Q. & Guo, S. W. Histological and Immunohistochemical Characterization of the Similarity and Difference Between Ovarian Endometriomas and Deep Infiltrating Endometriosis. Reprod Sci, 1933719117718275, https://doi.org/10.1177/1933719117718275 (2017). source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    SP or CGRP secreted by sensory nerves promote EMT-like morphological and molecular changes, and extended exposure of endometriotic epithelial cells to SP or CGRP also promotes myofibroblast activation.

    “Thus, we provided another piece of evidence that SP or CGRP secreted by sensory nerves promote EMT-like morphological and molecular changes, and extended exposure of endometriotic epithelial cells to SP or CGRP also promote myofibroblast activation.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: SP and CGRP are secreted mostly from sensory nerves.

  • May combine two claims.

    Interpretationown data

    SP and/or CGRP secreted from sensory nerves induce FMT and further SMM in endometriotic stromal cells.

    “These results strongly indicate that SP and/or CGRP secreted from sensory nerves induce FMT and further SMM in endometriotic stromal cells.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: SP and CGRP are secreted mostly from sensory nerves.

  • May combine two claims.

    Interpretationown data

    Sensory nerve-derived neuropeptides SP and CGRP facilitate EMT, FMT and differentiation to SMC in endometriosis, yielding increased collagen production, elevated cellular contractility, and eventually fibrosis.

    “In this study, we have shown that, as with platelet-derived TGF-β122, sensory nerve-derived neuropeptides SP and CGRP facilitate EMT, FMT and differentiation to SMC in endometriosis, yielding increased collagen production, elevated cellular contractility, and eventually fibrosis.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [122] Chernova, I. et al. Substance P (SP) enhances CCL5-induced chemotaxis and intracellular signaling in human monocytes, which express the truncated neurokinin-1 receptor (NK1R). Journal of leukocyte biology 85, 154–164, https://doi.org/10.1189/jlb.0408260 (2009). source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    SP plays an important role in cardiac fibrosis in response to hypertension.

    “SP plays an important role in cardiac fibrosis in response to hypertension80.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [80] Dehlin, H. M., Manteufel, E. J., Monroe, A. L., Reimer, M. H. Jr. & Levick, S. P. Substance P acting via the neurokinin-1 receptor regulates adverse myocardial remodeling in a rat model of hypertension. International journal of cardiology 168, 4643–4651, https://doi.org/10.1016/j.ijcard.2013.07.190 (2013). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    SP can directly induce M2 polarization of inflammatory macrophages, which participate in tissue repair and promote lesion growth and fibrogenesis in endometriosis.

    “SP can also directly induce M2 polarization of inflammatory macrophages, which participate in tissue repair75 and are involved in promoting lesion growth and fibrogenesis in endometriosis76,77.”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [75] Lim, J. E., Chung, E. & Son, Y. A neuropeptide, Substance-P, directly induces tissue-repairing M2 like macrophages by activating the PI3K/Akt/mTOR pathway even in the presence of IFNgamma. Sci Rep 7, 9417, https://doi.org/10.1038/s41598-017-09639-7 (2017). source

      Not checked in the free run.

    • cites [76] Bacci, M. et al. Macrophages are alternatively activated in patients with endometriosis and required for growth and vascularization of lesions in a mouse model of disease. Am J Pathol 175, 547–556, S0002-9440(10)60569-X (2009). source

      Not checked in the free run.

    • cites [77] Duan, J., Liu, X. & Guo, S.-W. The M2a macrophage subset may be critically involved in fibrogenesis of endometriosis in mouse. Reprod Biomed Online 37, 254–268, https://doi.org/10.1016/j.rbmo.2018.05.017. (2018). source

      Not checked in the free run.

  • May combine two claims.

    Interpretationreasoning

    Sensory nerve fibers and their secreted neuropeptides accelerate lesional development through facilitating EMT, FMT and SMM and ultimately fibrosis, making DE lesions typically more fibromuscular than OE lesions.

    “Considering the consensus that DE patients suffer much more serious pain, these results may explain the differences between OE and DE that sensory nerve fibers and their secreted neuropeptides accelerate the lesional development through facilitating EMT, FMT and SMM and ultimately fibrosis, making the DE lesions that are typically more fibromuscular than that of OE lesions.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: DE patients suffer much more serious pain than patients with other endometriosis subtypes.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Subcutaneous administration of SP accelerates the normal acute wound healing response via increased angiogenesis resulting from SP-mediated EPC mobilization.

    “Subcutaneous administration of SP accelerates the normal acute wound healing response via increased angiogenesis, resulting from SP-mediated EPC mobilization57.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [57] Um, J. et al. Substance P enhances EPC mobilization for accelerated wound healing. Wound Repair Regen 24, 402–410, https://doi.org/10.1111/wrr.12403 (2016). source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Sensory denervation impairs cutaneous wound healing through increased apoptosis and reduced proliferation.

    “It is well documented that sensory denervation impairs cutaneous wound healing through increased apoptosis and reduced proliferation50,51.”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [50] Smith, P. G. & Liu, M. Impaired cutaneous wound healing after sensory denervation in developing rats: effects on cell proliferation and apoptosis. Cell Tissue Res 307, 281–291, https://doi.org/10.1007/s00441-001-0477-8 (2002). source

      Not checked in the free run.

    • cites [51] Engin, C., Demirkan, F., Ayhan, S., Atabay, K. & Baran, N. K. Delayed effect of denervation on wound contraction in rat skin. Plast Reconstr Surg 98, 1063–1067 (1996). source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Successful repair of injured tissues requires diverse interactions among different cells, biochemical mediators, and the cellular microenvironment.

    “Successful repair of injured tissues requires diverse interactions among different cells, biochemical mediators, and the cellular microenvironment47–49.”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [47] Barbul, A. Immune aspects of wound repair. Clin Plast Surg 17, 433–442 (1990). source

      Not checked in the free run.

    • cites [48] Cioffi, W. G., Burleson, D. G. & Pruitt, B. A. Jr. Leukocyte responses to injury. Arch Surg 128, 1260–1267 (1993). source

      Not checked in the free run.

    • cites [49] Guirao, X. & Lowry, S. F. Biologic control of injury and inflammation: much more than too little or too late. World J Surg 20, 437–446 (1996). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Expression of NK1R and SP-coding gene TAC1 is upregulated by estrogen and TNF-α.

    “The expression of NK1R and SP-coding gene TAC1 is reported to be upregulated by estrogen68,71 and TNF-α44.”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [68] Pinto, F. M. et al. Tachykinin receptor and neutral endopeptidase gene expression in the rat uterus: characterization and regulation in response to ovarian steroid treatment. Endocrinology 140, 2526–2532, https://doi.org/10.1210/endo.140.6.6695 (1999). source

      Not checked in the free run.

    • cites [71] Pinto, F. M., Pintado, C. O., Pennefather, J. N., Patak, E. & Candenas, L. Ovarian steroids regulate tachykinin and tachykinin receptor gene expression in the mouse uterus. Reprod Biol Endocrinol 7, 77, https://doi.org/10.1186/1477-7827-7-77 (2009). source

      Not checked in the free run.

    • cites [44] McKinnon, B. D., Evers, J., Bersinger, N. A. & Mueller, M. D. Induction of the neurokinin 1 receptor by TNFalpha in endometriotic tissue provides the potential for neurogenic control over endometriotic lesion growth. J Clin Endocrinol Metab 98, 2469–2477, https://doi.org/10.1210/jc.2013-1019 (2013). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    NK1R is expressed in human and rodent uterus, as are CRLR and RAMP1.

    “NK1R is expressed in human and rodent uterus66–69, so are CRLR and RAMP170.”
    Quote in the paperSubstantial claim0 of 5 sources checked
    • cites [66] Patak, E. et al. Tachykinins and tachykinin receptors in human uterus. Br J Pharmacol 139, 523–532, https://doi.org/10.1038/sj.bjp.0705279 (2003). source

      Not checked in the free run.

    • cites [67] Patak, E. et al. Functional and molecular characterization of tachykinins and tachykinin receptors in the mouse uterus. Biol Reprod 72, 1125–1133, https://doi.org/10.1095/biolreprod.104.036814 (2005). source

      Not checked in the free run.

    • cites [68] Pinto, F. M. et al. Tachykinin receptor and neutral endopeptidase gene expression in the rat uterus: characterization and regulation in response to ovarian steroid treatment. Endocrinology 140, 2526–2532, https://doi.org/10.1210/endo.140.6.6695 (1999). source

      Not checked in the free run.

    • cites [69] Gonzalez-Santana, A. et al. Altered expression of the tachykinins substance P/neurokinin A/hemokinin-1 and their preferred neurokinin 1/neurokinin 2 receptors in uterine leiomyomata. Fertil Steril 106, 1521–1529, https://doi.org/10.1016/j.fertnstert.2016.07.007 (2016). source

      Not checked in the free run.

    • cites [70] Nikitenko, L. L. et al. Differential and cell-specific expression of calcitonin receptor-like receptor and receptor activity modifying proteins in the human uterus. Mol Hum Reprod 7, 655–664 (2001). source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Genetic deletion or blockade of CGRP receptors is detrimental to wound healing.

    “Similarly, genetic deletion or blockade of CGRP receptors has been suggested to be detrimental to wound healing58–60.”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [58] Brain, S. D. Sensory neuropeptides: their role in inflammation and wound healing. Immunopharmacology 37, 133–152 (1997). source

      Not checked in the free run.

    • cites [59] Brain, S. D. & Grant, A. D. Vascular actions of calcitonin gene-related peptide and adrenomedullin. Physiol Rev 84, 903–934, https://doi.org/10.1152/physrev.00037.2003 (2004). source

      Not checked in the free run.

    • cites [60] Toda, M. et al. Roles of calcitonin gene-related peptide in facilitation of wound healing and angiogenesis. Biomed Pharmacother 62, 352–359, https://doi.org/10.1016/j.biopha.2008.02.003 (2008). source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    SP accelerates normal acute and chronic wound healing processes.

    “SP accelerates the normal acute and chronic wound healing processes54–56.”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [54] Kant, V. et al. Topical application of substance P promotes wound healing in streptozotocin-induced diabetic rats. Cytokine 73, 144–155, https://doi.org/10.1016/j.cyto.2014.12.015 (2015). source

      Not checked in the free run.

    • cites [55] Leal, E. C. et al. Substance P promotes wound healing in diabetes by modulating inflammation and macrophage phenotype. Am J Pathol 185, 1638–1648, https://doi.org/10.1016/j.ajpath.2015.02.011 (2015). source

      Not checked in the free run.

    • cites [56] Yang, L. et al. Substance P promotes diabetic corneal epithelial wound healing through molecular mechanisms mediated via the neurokinin-1 receptor. Diabetes 63, 4262–4274, https://doi.org/10.2337/db14-0163 (2014). source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    SP mobilizes mesenchymal stem cells and endothelial progenitor cells in bone marrow and induces them to migrate into injured peripheral tissues where they are involved in tissue regeneration.

    “Furthermore, SP mobilizes mesenchymal stem cells52 and endothelial progenitor cells (EPCs)53 in the bone marrow, and induces them to migrate into the injured peripheral tissues where they are involved in tissue regeneration.”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [52] Hong, H. S. et al. A new role of substance P as an injury-inducible messenger for mobilization of CD29(+) stromal-like cells. Nat Med 15, 425–435, nm.1909 (2009). source

      Not checked in the free run.

    • cites [53] Amadesi, S. et al. Role for substance p-based nociceptive signaling in progenitor cell activation and angiogenesis during ischemia in mice and in human subjects. Circulation 125(1774–1786), S1771–1719, https://doi.org/10.1161/CIRCULATIONAHA.111.089763 (2012). source

      Not checked in the free run.

  • Classified as probably not a substantial claim. May combine two claims.

    Premisecitation

    Co-culture with rat primary sensory neurons from DRG as well as SP, CGRP and VIP facilitate fibroblast and keratinocyte proliferation and induce collagen I production.

    “Co-culture with rat primary sensory neurons from DRG and as well as SP, CGRP and vasoactive intestinal peptide or VIP facilitate fibroblasts and keratinocytes proliferation and induce collagen I production81.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [81] Cheret, J. et al. Influence of sensory neuropeptides on human cutaneous wound healing process. Journal of dermatological science 74, 193–203, https://doi.org/10.1016/j.jdermsci.2014.02.001 (2014). source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Direct contact of fibroblasts with neuronal processes promotes differentiation to myofibroblasts and their contractility.

    “Direct contact of fibroblasts with neuronal processes promotes differentiation to myofibroblasts and their contractility82.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [82] Fujiwara, T. et al. Direct contact of fibroblasts with neuronal processes promotes differentiation to myofibroblasts and induces contraction of collagen matrix in vitro. Wound Repair Regen 21, 588–594, https://doi.org/10.1111/wrr.12059 (2013). source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    SP acts as an immune modulator and injury messenger in various peripheral tissues.

    “Indeed, SP acts as an immune modulator and injury messenger in various peripheral tissues52.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [52] Hong, H. S. et al. A new role of substance P as an injury-inducible messenger for mobilization of CD29(+) stromal-like cells. Nat Med 15, 425–435, nm.1909 (2009). source

      Not checked in the free run.

  • May combine two claims.

    Interpretationreasoning

    The results presented in this and the companion manuscript are consistent with the roles of SP/CGRP and their receptors in wound healing and fibrogenesis, and support that sensory nerves are a notable feature of the lesional microenvironment, especially in DE lesions, and that innervation plays an important role in lesional development and fibrogenesis.

    “In view of the above, the results presented in this and the companion manuscript are consistent with the roles of SP/CGRP and their receptors in wound healing and fibrogenesis, and underscores the notion that sensory nerves are a notable feature of the lesional microenvironment, especially in DE lesions, and innervation plays an important role in lesional development and fibrogenesis.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    SP has been reported to be a potent neuroimmunomodulator functioning through NK1R, which is widely expressed in immune cells.

    “Functioning through NK1R, which is widely expressed in immune cells, SP has been reported to be a potent neuroimmunomodulator78.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [78] Mashaghi, A. et al. Neuropeptide substance P and the immune response. Cell Mol Life Sci 73, 4249–4264, https://doi.org/10.1007/s00018-016-2293-z. source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Interpretationcitation

    The results are consistent with the well documented roles of SP/CGRP and their receptors in wound healing and fibrogenesis.

    “Viewed from the lens of the ReTIAR22, our results are consistent with the well documented roles of SP/CGRP and their receptors in wound healing and fibrogenesis.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [22] Zhang, Q., Duan, J., Liu, X. & Guo, S. W. Platelets drive smooth muscle metaplasia and fibrogenesis in endometriosis through epithelial-mesenchymal transition and fibroblast-to-myofibroblast transdifferentiation. Mol Cell Endocrinol 428, 1–16, S0303-7207(16)30063-6 (2016). source

      Not checked in the free run.

  • May combine two claims.

    Interpretationreasoning

    Increased local production of estrogen and proinflammatory cytokines may induce NK1R in myometrium, causing uterine hyperactivity and subsequently pain.

    “This may suggest that increased local production of estrogen and proinflammatory cytokines may induce NK1R in myometrium, causing uterine hyperactivity and, subsequently, pain, as in colon72,73.”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [72] Bradesi, S., Eutamene, H., Fioramonti, J. & Bueno, L. Acute restraint stress activates functional NK1 receptor in the colon of female rats: involvement of steroids. Gut 50, 349–354 (2002). source

      Not checked in the free run.

    • cites [73] Lu, P. et al. The role of substance P in the maintenance of colonic hypermotility induced by repeated stress in rats. Neuropeptides 56, 75–82, https://doi.org/10.1016/j.npep.2016.01.006 (2016). source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

    Premisenone given

    All these proteins have been implicated in the development of endometriosis.

    “Importantly, all these proteins have been implicated in the development of endometriosis.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationreasoning

    Platelet-derived TGF-β1, macrophages, regulatory T cells, and sensory nerve-derived neuropeptides can all induce EMT, FMT, and SMM, resulting in lesional fibrosis.

    “While in each case the identity of the culprit in promoting lesional fibrogenesis is different, all of them nonetheless can induce EMT, FMT, and SMM, resulting in ultimately lesional fibrosis.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    NK1R antagonism can mitigate alterations in the brain induced by chronic psychological stress, which is associated with visceral hypersensitivity and spinal NK1R up-regulation in female rodents and possibly in mice with induced endometriosis.

    “Aside from delaying fibrogenesis, NK1R antagonism can also mitigate alterations in the brain induced by chronic psychological stress99–101, which is associated with visceral hypersensitivity and spinal NK1R up-regulation in female rodents102,103 and possibly in mouse with induced endometriosis as well104–106.”
    Quote in the paperNot a substantial claim0 of 5 sources checked
    • cites [99] Czeh, B. et al. Examining SLV-323, a novel NK1 receptor antagonist, in a chronic psychosocial stress model for depression. Psychopharmacology 180, 548–557, https://doi.org/10.1007/s00213-005-2184-8 (2005). source

      Not checked in the free run.

    • cites [100] Czeh, B. et al. Chronic stress decreases the number of parvalbumin-immunoreactive interneurons in the hippocampus: prevention by treatment with a substance P receptor (NK1) antagonist. Neuropsychopharmacology 30, 67–79, https://doi.org/10.1038/sj.npp.1300581 (2005). source

      Not checked in the free run.

    • cites [101] van der Hart, M. G. et al. Substance P receptor antagonist and clomipramine prevent stress-induced alterations in cerebral metabolites, cytogenesis in the dentate gyrus and hippocampal volume. Molecular psychiatry 7, 933–941, https://doi.org/10.1038/sj.mp.4001130 (2002). source

      Not checked in the free run.

    • cites [102] Bradesi, S., Eutamene, H., Garcia-Villar, R., Fioramonti, J. & Bueno, L. Stress-induced visceral hypersensitivity in female rats is estrogen-dependent and involves tachykinin NK1 receptors. Pain 102, 227–234 (2003). source

      Not checked in the free run.

    • cites [103] Bradesi, S. et al. Role of spinal microglia in visceral hyperalgesia and NK1R up-regulation in a rat model of chronic stress. Gastroenterology 136, 1339–1348, e1331–1332, https://doi.org/10.1053/j.gastro.2008.12.044 (2009). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    DE lesions are typically in close proximity to various sensory nerve plexuses and have higher density of sensory nerves than other subtypes.

    “The answer simply lies in the locations of the DE lesions: unlike other subtypes, these lesions typically are in close proximity to various sensory nerve plexuses and additionally have higher density of sensory nerves than other subtypes29,30,85,86.”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [29] Wang, G., Tokushige, N., Markham, R. & Fraser, I. S. Rich innervation of deep infiltrating endometriosis. Hum Reprod 24, 827–834, den464 (2009). source

      Not checked in the free run.

    • cites [30] Wang, G. et al. Hyperinnervation in intestinal deep infiltrating endometriosis. J Minim Invasive Gynecol 16, 713–719, S1553-4650(09)00357-4 (2009). source

      Not checked in the free run.

    • cites [85] Tokushige, N., Markham, R., Russell, P. & Fraser, I. S. Nerve fibres in peritoneal endometriosis. Hum Reprod 21, 3001–3007, del260 (2006). source

      Not checked in the free run.

  • May combine two claims.

    Interpretationreasoning

    Sensory nerves in or around endometriotic lesions contribute to pain in women with endometriosis through accelerated lesional fibrogenesis and enhanced transduction of pain mediators to the CNS.

    “However, the sensory nerves in or around endometriotic lesions also become accessories to the crime in inflicting pains in women with endometriosis through accelerated lesional fibrogenesis and enhanced transduction of pain mediators to the CNS.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Study design and conductnone given

    The study adhered to Helsinki Declaration ethical principles and was approved by the institutional ethics review board of Shanghai OB/GYN Hospital, Fudan University.

    “This study strictly adhered to the ethical principles outlined by the Helsinki Declaration and was approved by the institutional ethics review board of Shanghai OB/GYN Hospital, Fudan University.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisecitation

    DE is quite different from OE or PE and its histology resembles that of uterine adenomyosis.

    “DE is quite different from OE or PE2 and its histology resembles that of uterine adenomyosis6.”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [2] Nisolle, M. & Donnez, J. Peritoneal endometriosis, ovarian endometriosis, and adenomyotic nodules of the rectovaginal septum are three different entities. Fertil Steril 68, 585–596, S0015-0282(97)00191-X (1997). source

      Not checked in the free run.

    • cites [6] Gordts, S., Koninckx, P. & Brosens, I. Pathogenesis of deep endometriosis. Fertil Steril 108, 872–885 e871, S0015-0282(17)31884-8 (2017). source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Interpretationreasoning

    The identical processes of EMT, FMT, SMM, and fibrogenesis instituted by platelets, immune cells and sensory nerves in endometriotic cells underscore the dynamic nature of cellular identity, phenotype, function and behavior of endometriotic cells.

    “In addition, the identical processes, instituted by platelets, immune cells and sensory nerves, of EMT, FMT, SMM, and fibrogenesis experienced by endometriotic cells underscore the dynamic nature in cellular identity, phenotype, function and behavior of endometriotic cells.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationcitation

    OE and DE, and likely other subtypes of endometriosis and adenomyosis, have identical pathophysiology and similar natural history as wounds undergoing ReTIAR.

    “As a corollary, OE and DE, and likely other subtypes of endometriosis and adenomyosis as well91,92, have the identical pathophysiology and similar natural history as wounds undergoing ReTIAR.”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [91] Liu, X., Shen, M., Qi, Q., Zhang, H. & Guo, S. W. Corroborating evidence for platelet-induced epithelial-mesenchymal transition and fibroblast-to-myofibroblast transdifferentiation in the development of adenomyosis. Hum Reprod 31, 734–749, dew018 (2016). source

      Not checked in the free run.

    • cites [92] Shen, M., Liu, X., Zhang, H. & Guo, S. W. Transforming growth factor beta1 signaling coincides with epithelial-mesenchymal transition and fibroblast-to-myofibroblast transdifferentiation in the development of adenomyosis in mice. Hum Reprod 31, 355–369, dev314 (2016). source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductnone given

    Written informed consent was obtained from all study subjects prior to sample collection.

    “Written informed consent was obtained from all study subjects prior to sample collection.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationreasoning

    The lesional microenvironment is important in lesional development and fibrogenesis, and endometriotic cells are dynamic.

    “Our study highlights the importance of lesional microenvironment in lesional development and fibrogenesis and also the dynamic nature of endometriotic cells.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Study design and conductnone given

    None of the patients had received hormonal or anti-platelet treatment for at least 90 days before surgery, and none had malignant or other inflammatory disease.

    “None of the patients had received hormonal or anti-platelet treatment for at least 90 days before the surgery. In addition, none of them had any malignant or other inflammatory disease.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationreasoning

    Sensory nerve-derived neuropeptides, which are abundant in the DE microenvironment, can turn non-DE endometriotic or even endometrial stromal cells into DE-like cells, resulting in a fibromuscular phenotype characteristic of DE.

    “This, in fact, is what this study is trying to show: sensory nerve-derived neuropeptides, which are abundant in the DE microenvironment, can turn non-DE endometriotic or even endometrial stromal cells into DE-like cells, resulting in a fibromuscular phenotype that is characteristically DE.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: SP and CGRP are abundant in the DE lesional microenvironment.

  • May combine two claims.

    Interpretationreasoning

    In the lesional microenvironment, several factors promote endometriosis and may conspire together or act independently to promote lesional fibrogenesis.

    “Hence in lesional microenvironment, there are several accessories to the crime of promoting endometriosis, who may conspire together (e.g., lesion- and/or platelet-derived TXA2 induces neurite outgrowth, and thus lesional innervation84) or may act independently, that together promote lesional fibrogenesis.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [84] Yan, D., Liu, X. & Guo, S. W. Endometriosis-Derived Thromboxane A2 Induces Neurite Outgrowth. Reprod Sci 24, 829–835, 1933719116670037 (2017). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Endometriosis can induce anxiety and depression in mice and possibly in humans.

    “Given that endometriosis can induce anxiety and depression in mouse108 and possibly in humans as well109–114, the use of NK1R inhibitors as a therapeutics for endometriosis may have added benefits.”
    Quote in the paperSubstantial claim0 of 7 sources checked
    • cites [108] Li, T. et al. Endometriosis alters brain electro-physiology, gene expression and increased pain sensitization, anxiety, and depression in female mice. Biol Reprod 4841840 (2018)

      Not checked in the free run.

    • cites [109] Low, W. Y., Edelmann, R. J. & Sutton, C. A psychological profile of endometriosis patients in comparison to patients with pelvic pain of other origins. J Psychosom Res 37, 111–116 (1993). source

      Not checked in the free run.

    • cites [110] Barnack, J. L. & Chrisler, J. C. The experience of chronic illness in women: a comparison between women with endometriosis and women with chronic migraine headaches. Women Health 46, 115–133, https://doi.org/10.1300/J013v46n01_08 (2007). source

      Not checked in the free run.

    • cites [111] Petrelluzzi, K. F., Garcia, M. C., Petta, C. A., Grassi-Kassisse, D. M. & Spadari-Bratfisch, R. C. Salivary cortisol concentrations, stress and quality of life in women with endometriosis and chronic pelvic pain. Stress 11, 390–397, 902547461 (2008). source

      Not checked in the free run.

    • cites [112] Eriksen, H. L. et al. Psychological aspects of endometriosis: differences between patients with or without pain on four psychological variables. Eur J Obstet Gynecol Reprod Biol 139, 100–105, S0301-2115(07)00436-8 (2008). source

      Not checked in the free run.

    • cites [113] Sepulcri Rde, P. & do Amaral, V. F. Depressive symptoms, anxiety, and quality of life in women with pelvic endometriosis. Eur J Obstet Gynecol Reprod Biol 142, 53–56, S0301-2115(08)00335-7 (2009). source

      Not checked in the free run.

    • cites [114] Quinones, M., Urrutia, R., Torres-Reveron, A., Vincent, K. & Flores, I. Anxiety, coping skills and hypothalamus-pituitary-adrenal (HPA) axis in patients with endometriosis. J Reprod Biol Health 3, https://doi.org/10.7243/2054-0841-3-2 (2015). source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductnone given

    Dorsal root ganglia were used as a positive control for CGRP staining.

    “Dorsal root ganglia were used as positive control for the CGRP staining.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Study design and conductown data

    The purity of primary endometriotic stromal cells was confirmed by positive vimentin and negative CK7 staining after the third passage, and contamination with ovarian granulosa cells was ruled out by negative follicle stimulating hormone receptor staining.

    “The vimentin staining was positive and the CK7 staining was negative after the third passage. To rule out the possibility of contamination with ovarian granulosa cells, we also stained the primary cells with follicle stimulating hormone receptor (Abcam, Cambridge, UK), a specific marker for granulosa cells118. The staining was found to be negative, as we previously reported119.”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [118] Lan, C. W., Chen, M. J., Jan, P. S., Chen, H. F. & Ho, H. N. Differentiation of human embryonic stem cells into functional ovarian granulosa-like cells. J Clin Endocrinol Metab 98, 3713–3723, https://doi.org/10.1210/jc.2012-4302 (2013). source

      Not checked in the free run.

    • cites [119] Zhang, Q., Ding, D., Liu, X. & Guo, S. W. Activated Platelets Induce Estrogen Receptor beta Expression in Endometriotic Stromal Cells. Gynecol Obstet Invest 80, 187–192, https://doi.org/10.1159/000377629 (2015). source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    The major receptors for calcitonin gene-related peptide (CGRP) are calcitonin receptor-like receptor (CRLR) and receptor activity modifying protein 1 (RAMP-1).

    “while for CGRP, its major receptors are calcitonin receptor like receptor (CRLR) and receptor activity modifying protein 1 (RAMP-1).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [134] McLatchie, L. M. et al. RAMPs regulate the transport and ligand specificity of the calcitonin-receptor-like receptor. Nature 393, 333–339, https://doi.org/10.1038/30666 (1998). source

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Study design and conductcitation

    All scratch assays were conducted in the presence of 5 μg/mL mitomycin-C to suppress cellular proliferation.

    “All experiments were conducted in the presence of 5 μg/mL of mitomycin-C (Sigma) to suppress cellular proliferation133.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [133] Lee, J. G. & Kay, E. P. FGF-2-induced wound healing in corneal endothelial cells requires Cdc42 activation and Rho inactivation through the phosphatidylinositol 3-kinase pathway. Invest Ophthalmol Vis Sci 47, 1376–1386, https://doi.org/10.1167/iovs.05-1223 (2006). source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    CRLR cannot function as a receptor for CGRP by itself; it requires complex formation with accessory proteins from the RAMP family.

    “CRLR by itself cannot function as a receptor for CGRP as it needs to form complexes with accessory proteins from the RAMP family134.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [134] McLatchie, L. M. et al. RAMPs regulate the transport and ligand specificity of the calcitonin-receptor-like receptor. Nature 393, 333–339, https://doi.org/10.1038/30666 (1998). source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductnone given

    All invasion experiments were carried out in triplicates.

    “All experiments were carried out in triplicates.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductnone given

    The first slide from each tissue block was stained with hematoxylin and eosin to validate the pathologic diagnosis.

    “The first resultant slide was stained with hematoxylin and eosin to validate pathologic diagnosis”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Study design and conductcitation

    The animal study followed the National Research Council's Guide for the Care and Use of Laboratory Animals and was approved by the Ethics Review Board, Shanghai OB/GYN Hospital.

    “Thirty virgin female Sprague-Dawley rats (Shanghai Center for Experimental Animals, Chinese Academy of Sciences, Shanghai, China), 4–5 weeks old, 100–120 g in bodyweight, were used for this study following the guidelines of the National Research Council’s Guide for the Care and Use of Laboratory Animals130 and approval by the Ethics Review Board, Shanghai OB/GYN Hospital.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [130] In Guide for the Care and Use of Laboratory Animals The National Academies Collection: Reports funded by National Institutes of Health (2011).

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductnone given

    Negative control sections were processed by omitting the primary antibody.

    “Negative control sections were processed by omitting the primary antibody.”
    Quote in the paperNot a substantial claim

  • The quote only partly states this claim.

    Study design and conductcitation

    The doses of SP, CGRP, aprepitant, and CGRP 8-37 used were justified based on prior reports and plasma concentrations.

    “With these considerations, we believe that our choice of doses are well justified.”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [120] Nilsson, J., von Euler, A. M. & Dalsgaard, C. J. Stimulation of connective tissue cell growth by substance P and substance K. Nature 315, 61–63 (1985). source

      Not checked in the free run.

    • cites [121] Liang, W., Zhuo, X., Tang, Z., Wei, X. & Li, B. Calcitonin gene-related peptide stimulates proliferation and osteogenic differentiation of osteoporotic rat-derived bone mesenchymal stem cells. Mol Cell Biochem 402, 101–110, https://doi.org/10.1007/s11010-014-2318-6 (2015). source

      Not checked in the free run.

    • cites [122] Chernova, I. et al. Substance P (SP) enhances CCL5-induced chemotaxis and intracellular signaling in human monocytes, which express the truncated neurokinin-1 receptor (NK1R). Journal of leukocyte biology 85, 154–164, https://doi.org/10.1189/jlb.0408260 (2009). source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductown data

    The CCK-8 experiment was done in triplicate.

    “The experiment was done in triplicate.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    The endogenous receptor for substance P is neurokinin 1 receptor (NK1R).

    “The endogenous receptor for SP is NK1R44”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [44] McKinnon, B. D., Evers, J., Bersinger, N. A. & Mueller, M. D. Induction of the neurokinin 1 receptor by TNFalpha in endometriotic tissue provides the potential for neurogenic control over endometriotic lesion growth. J Clin Endocrinol Metab 98, 2469–2477, https://doi.org/10.1210/jc.2013-1019 (2013). source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductown data

    Cell immunofluorescence experiments were carried out in duplicate.

    “All experiments were carried out in duplicate.”
    Quote in the paperNot a substantial claim
62 of 62

  • The paper relies on this without support.

    DE lesions are in close proximity to several nerve plexuses such as inferior hypogastric, vesical, uterovaginal, and rectal plexus.

    “In contrast to OE lesions, DE lesions are in uncanny proximity to several nerve plexuses such as inferior hypogastric, vesical, uterovaginal, and rectal plexus.”

  • The paper relies on this without support.

    SP and CGRP are secreted mostly from sensory nerves.

    “Considering that SP and/or CGRP, which are secreted mostly from sensory nerves, induced FMT and SMM in endometriotic stromal cells as shown above”

  • The paper relies on this without support.

    DE patients suffer much more serious pain than patients with other endometriosis subtypes.

    “Considering the consensus that DE patients suffer much more serious pain”

  • The paper relies on this without support.

    SP and CGRP are abundant in the DE lesional microenvironment.

    “sensory nerve-derived neuropeptides, which are abundant in the DE microenvironment”
4 of 4

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

Each contradiction between two passages listed here was confirmed by an independent check.

  • Figure 7 collagen-assay duration conflicts with Methods

    Major

    Figure 7D reports collagen production after 12 days of treatment, but the collagen-assay method says culture medium was collected after 72 hours of treatment.

  • CCK-8 treatment duration differs between Results and Methods

    Minor

    The Results describe the 11Z proliferation assay after 72 hours of treatment; the CCK-8 method says the assay solution was added after 48 hours.

  • Figure 3B is described as both protein and gene expression

    Minor

    The Results cite Figure 3B as evidence of increased protein expression, but the figure caption says panel B reports relative gene-expression levels.

3 of 3

Cites

Flagged first, then by citations

1 of the 136 references couldn’t be matched to a known paper.

Research works printed but not linked: 2

  1. Liu, X., Yan, D. & Guo, S. W. Establishment of a mouse model of deep infiltrating endometriosis [Abstract]. Reprod Sci 25, 61A (2018).
  2. Zhang, Q., Liu, X. & Guo, S. W. Progressive development of endometriosis and its hindrance by anti-platelet treatment in mice with induced endometriosis. Reproductive biomedicine online, https://doi.org/10.1016/j.rbmo.2016.11.006 (2016). · DOI · Paper page

Cited by

Sorted by citations

Related

Authors
Yan, D., Liu, X., & Guo, S.-W.
Affiliations
Shanghai Medical College of Fudan University (CN)
Corresponding author
Guo, Sun-Wei
ORCID
Venue
Scientific Reports
Publisher
Springer Science and Business Media LLC
Published
2019-02-25
Volume
9
Issue
1
Pages
2698-2698
ISSN
2045-2322 (ISSN-L)
DOI
10.1038/s41598-019-39170-w
PMC
PMC6389969
OpenAlex
W2917664025
Semantic Scholar
7a4351683f8c5649b2eb4506fd1cfd7c3f1caf2d
Cited by
82
References
136
Citations by year
2026: 9 · 2025: 9 · 2024: 11 · 2023: 10 · 2022: 12 · 2021: 17 · 2020: 10 · 2019: 4
FWCI
10.28
Type
journal article
Language
en
Open access
Licence
Locations
  • Scientific Reports (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, other-oa)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
Funders
National Natural Science Foundation of China (81671436, 81871144, 81771553)
Topics
  • Health Sciences › Medicine › Reproductive Medicine › Endometriosis Research and Treatment
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuropeptides and Animal Physiology
  • Life Sciences › Neuroscience › Sensory Systems › Ion Channels and Receptors
Keywords
endometriosis, deep infiltrating endometriosis, substance P, calcitonin gene-related peptide, neuropeptides, fibrosis, epithelial-mesenchymal transition, fibroblast-to-myofibroblast transdifferentiation, smooth muscle differentiation, sensory nerves, dorsal root ganglia, calcitonin receptor-like receptor, endometriotic stromal cells, nerve innervation, endometrioma
MeSH terms
Animals, Cells, Cultured, Endometriosis, Female, Fibroblasts, Immunohistochemistry, Substance P, Blotting, Western, Calcitonin Gene-Related Peptide, Rats, Sprague-Dawley, Receptors, Neurokinin-1, Cell Proliferation, Rats, Myofibroblasts, Epithelial-Mesenchymal Transition

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-10.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-10.

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