AMP-activated protein kinase is involved in endothelial NO synthase activation in response to shear stress
Abstract
OBJECTIVE: The regulation of AMP-activated protein kinase (AMPK) is implicated in vascular biology because AMPK can phosphorylate endothelial NO synthase (eNOS). In this study, we investigate the regulation of the AMPK-eNOS pathway in vascular endothelial cells (ECs) by shear stress and the activation of aortic AMPK in a mouse model with a high level of voluntary running (High-Runner). METHODS AND RESULTS: By using flow channels with cultured ECs, AMPK Thr172 phosphorylation was increased with changes of flow rate or pulsatility. The activity of LKB1, the upstream kinase of AMPK, and the phosphorylation of eNOS at Ser1179 were concomitant with AMPK activation responding to changes in flow rate or pulsatility. The blockage of AMPK by a dominant-negative mutant of AMPK inhibited shear stress-induced eNOS Ser1179 phosphorylation and NO production. Furthermore, aortic AMPK activity and level of eNOS phosphorylation were significantly elevated in the aortas of High-Runner mice. CONCLUSIONS: Our results suggest that shear stress activates AMPK in ECs, which contributes to elevated eNOS activity and subsequent NO production. Hence, AMPK, in addition to serving as an energy sensor, also plays an important role in regulating vascular tone.
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Similar general outcome of metformin, AICAR, atheroprotective flow, and statins on endothelial cells can be found through AMPK activation.
“As discussed previously, a similar general outcome of metformin, AICAR, atheroprotective flow, and statins on ECs can be found through AMPK activation [245,250,251].”
From AMPK, Mitochondrial Function, and Cardiovascular Disease (2020) · cites this paper as [245]
Metformin has been proven as an anti-inflammatory, especially for the inflammation in diabetes and following postsurgical endotoxin.
“Metformin has been proven as an anti-inflammatory, especially for the inflammation in diabetes and following postsurgical endotoxin [245,246,247].”
From AMPK, Mitochondrial Function, and Cardiovascular Disease (2020) · cites this paper as [245]
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“Nitrite levels are increased following exercise in both rodents and humans [110, 111].”
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“Furthermore, both of the amino acid sequences flanking Ser-27 and Ser-47 do not align well with the phosphorylation consensus sequence of LKB1 (Fig.”
BackgroundInfluentialAcute exercise activates AMPK and eNOS in the mouse aorta
Cacicedo, J. M., Gauthier, M., LeBrasseur, N. K., Jasuja, R., Ruderman, N. B., & Ido, Y. (2011) · American Journal of Physiology-Heart and Circulatory Physiology · doi:10.1152/ajpheart.01279.2010 · Cited by 81 · see citing works
Targeting AMPK for cardiac protection: Opportunities and challenges
Kim, M., & Tian, R. (2010) · Journal of Molecular and Cellular Cardiology · doi:10.1016/j.yjmcc.2010.12.004 · Cited by 82 · see citing works
“Previous studies showed that AMPK activated eNOS (endothelial NO synthase) in endothelial cells in response to shear stress and myocardial ischemia contributing to the regulation of vascular tone under these condtions [73,74].”
BackgroundAtheroprotective laminar flow inhibits Hippo pathway effector YAP in endothelial cells
Xu, S., Koroleva, M., Yin, M., & Jin, Z. G. (2016) · Translational research · doi:10.1016/j.trsl.2016.05.003 · Cited by 81 · see citing works
“1022 1023 Ser127, and other sites, Ser94 and Ser61 in particular, because 3 independent research groups recently reported glucose deprivation–induced energy stress activates LATS kinase and AMPK activation followed by YAP phosphorylation and inactivation,(32-34) and it has been shown that LF increases AMPK activity.(3…”
BackgroundPerivascular fat, AMP ‐activated protein kinase and vascular diseases
Almabrouk, T. A. M., Ewart, M., Salt, I. P., & Kennedy, S. (2013) · British Journal of Pharmacology · doi:10.1111/bph.12479 · Cited by 80 · see citing works
“In eNOS knockout mice, shear stress- and bradykinin-mediated activation of AMPK is attenuated (Zhang et al., 2006; Wang et al., 2009a) and pharmacological inhibition of NOS prevents activation of AMPK by shear stress, bradykinin and calcium ionophores (Zhang et al.”
BackgroundCastiglione, N., Rinaldo, S., Giardina, G., Stelitano, V., & Cutruzzolà, F. (2012) · Antioxidants and Redox Signaling · doi:10.1089/ars.2011.4196 · Cited by 79 · see citing works
“The levels of nitrite in the body show significant variability due to differences in dietary habits, lifestyle ( e.g. , tobacco consumption), and physical exercise (37, 255, 265, 375).”
Fißlthaler, B., Fleming, I., Keserü, B., Walsh, K., & Busse, R. (2007) · Circulation Research · doi:10.1161/01.res.0000257747.74358.1c · Cited by 75 · see citing works
Bendale, D. S., Karpe, P. A., Chhabra, R., Shete, S. P., Shah, H., & Tikoo, K. (2013) · British Journal of Pharmacology · doi:10.1111/bph.12290 · Cited by 74 · see citing works
“Recently the increase in expression of P-AMPK and SIRT1 has been positively correlated with eNOS phosphorylation, NO bioavailability and endothelial function (Zhang et al., 2006; Mattagajasingh et al., 2007).”
BackgroundChen, W., Chen, C.-H., Hsu, M., Chang, R.-W., Wang, C., & Lee, T. (2024) · Biomedicine & Pharmacotherapy · doi:10.1016/j.biopha.2024.116192 · Cited by 70 · see citing works
Adam, J., Brandmaier, S., Leonhardt, J., Scheerer, M. F., Mohney, R. P., Xu, T., Bi, J., Rotter, M., Troll, M., Chi, S., et al. (2016) · Diabetes · doi:10.2337/db16-0512 · Cited by 68 · see citing works
“AMPK in turn may stimulate eNOS by its phosphorylation (8,41), which suggests a consequent increase of the NO production in the NO cycle (Fig.”
BackgroundA novel TRPV4-specific agonist inhibits monocyte adhesion and atherosclerosis
Xu, S., Liu, B., Yin, M., Королева, М. В., Mastrangelo, M. A., Ture, S. K., Morrell, C. N., Zhang, D. X., Fisher, E. A. A., & Jin, Z. G. (2016) · Oncotarget · doi:10.18632/oncotarget.9376 · Cited by 68 · see citing works
“A, GSK1016790A (GSK101) increases AMPK activation.”
InfluentialRodríguez, C., Contreras, C., Medina, J. S., Muñoz, M., Corbacho, C., Carballido, J., García‐Sacristán, A., Hernández, M., López, M., Rivera, L., et al. (2020) · Redox Biology · doi:10.1016/j.redox.2020.101575 · Cited by 68 · see citing works
Protein kinase N2 mediates flow-induced endothelial NOS activation and vascular tone regulation
Jin, Y., Chennupati, R., Li, R., Liang, G., Wang, S., Iring, A., Graumann, J., Wettschureck, N., & Offermanns, S. (2021) · Journal of Clinical Investigation · doi:10.1172/jci145734 · Cited by 67 · see citing works
“Later, other related protein kinases, including AMP-activated kinase (AMPK) and PKA have been reported to be also able to mediate flow-induced eNOS phosphorylation at serine 1177 (4, 5, 13, 14).”
BackgroundMackenzie, R. M., Salt, I. P., Miller, W. H., Logan, A., Ibrahim, H., Degasperi, A., Dymott, J. A., Hamilton, C. A., Murphy, M. P., Delles, C., et al. (2012) · Clinical Science · doi:10.1042/cs20120239 · Cited by 67 · see citing works
Beneficial and Detrimental Roles of Heme Oxygenase-1 in the Neurovascular System
Choi, Y. K., & Kim, Y. (2022) · International Journal of Molecular Sciences · doi:10.3390/ijms23137041 · Cited by 63 · see citing works
“In experiments on flow channels in cultured endothelial cells, AMPKα phosphorylation was increased with changes in the flow rate or pulsatility [91].”
BackgroundChatauret, N. P., Coudroy, R., Delpech, P., Vandebrouck, C., Hosni, S., Scépi, M., & Hauet, T. (2014) · American Journal of Transplantation · doi:10.1111/ajt.12904 · Cited by 64 · see citing works
Wu, Y., Zhang, C., Dong, Y., Wang, S., Song, P., Viollet, B., & Zou, M. (2012) · PLoS ONE · doi:10.1371/journal.pone.0035508 · Cited by 64 · see citing works
“[28] demonstrated that infection of endothelial cells with a recombinant adenovirus expressing the constitutively active AMPK results in eNOS activation and increased NO production.”
BackgroundYalçın, Ö., Ülker, P., Yavuzer, U., Meiselman, H. J., & Baskurt, O. K. (2008) · American Journal of Physiology-Heart and Circulatory Physiology · doi:10.1152/ajpheart.00015.2008 · Cited by 64 · see citing works
“on February 15, 2010 ajpheart.physiology.org Downloaded from synthesis (30) and eNOS phosphorylation (7, 48 ).”
BackgroundPenumathsa, S. V., Thirunavukkarasu, M., Samuel, S. M., Zhan, L., Maulik, G., Bagchi, M., Bagchi, D., & Maulik, N. (2008) · Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease · doi:10.1016/j.bbadis.2008.10.018 · Cited by 63 · see citing works
Bradley, E. A., Eringa, E. C., Stehouwer, C. D. A., Korstjens, I. J. M., Amerongen, G. P. V. N., Musters, R. J. P., Sipkema, P., Clark, M. G., & Rattigan, S. (2010) · Arteriosclerosis Thrombosis and Vascular Biology · doi:10.1161/atvbaha.110.204404 · Cited by 62 · see citing works
AMPKα2 exerts its anti-inflammatory effects through PARP-1 and Bcl-6
Gongol, B., Marin, T., Peng, I.-C., Woo, B., Martin, M., King, S., Sun, W., Johnson, D. A., Chien, S., & Shyy, J. Y.-J. (2013) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1222051110 · Cited by 61 · see citing works
“In particular, pulsatile flow (PS) activates AMPK (25) and is associated with anti-inflammatory effects (6– 9).”
BackgroundPark, J., Lee, S., Cho, D., Park, Y. M., Kang, D., & Jo, I. (2013) · Biochemical and Biophysical Research Communications · doi:10.1016/j.bbrc.2013.06.003 · Cited by 59 · see citing works
Xu, L., Wang, S., Li, B., Sun, A., Zou, Y., & Ge, J. (2014) · Journal of Cellular and Molecular Medicine · doi:10.1111/jcmm.12463 · Cited by 55 · see citing works
“Metformin, the AMPK activator, proved to be effective in meditating vascular protection against calcification, improving myocardial infarction and attenuating ventricular hypertrophy, all of which required the mechanism of AMPK-dependent eNOS activation [26, 29, 42, 43].”
BackgroundXu, Q., Hao, X., Yang, Q., & Si, L. (2009) · Biochemical and Biophysical Research Communications · doi:10.1016/j.bbrc.2009.08.021 · Cited by 56 · see citing works
Ching, L., Chen, C., Su, K., Hou, H., Shyue, S., Kou, Y. R., & Lee, T. (2012) · Molecular Medicine · doi:10.2119/molmed.2011.00461 · Cited by 54 · see citing works
“For instance, shear stress, the major physiological stimulation in the vascular system, was found to activate AMPK, which in turn phosphorylated eNOS at Ser635 and Ser1179, thus leading to enhanced NO bioavailability (27,36).”
BackgroundAtheroprone flow enhances the endothelial-to-mesenchymal transition
Lai, B., Li, Z., He, M., Wang, Y., Chen, L., Zhang, J., Yang, Y., & Shyy, J. Y.-J. (2018) · American Journal of Physiology-Heart and Circulatory Physiology · doi:10.1152/ajpheart.00213.2018 · Cited by 53 · see citing works
Han, Y., He, M., Marin, T., Shen, H., Wang, W., Lee, T.-Y., Hong, H.-C., Jiang, Z., Garland, T., Shyy, J. Y.-J., et al. (2021) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.2103982118 · Cited by 53 · see citing works
“We used adult males from a selectively bred mouse model (generation 83) with a high level of voluntary wheel running, namely, high-runner (HR) mice (28), to substantiate the finding that KLF4-AMPK/GCKR inhibits glycolysis in the endothelium in vivo.”
BackgroundMethodInfluentialFord, R. J., & Rush, J. W. E. (2010) · American Journal of Physiology-Heart and Circulatory Physiology · doi:10.1152/ajpheart.00597.2010 · Cited by 53 · see citing works
“Although recognized primarily as a modulator of cellular energy status and metabolism (28, 57), the identification of its existence in vascular cells and of its ability to be stimulated by numerous physical (8, 20, 59), energetic (17), hormonal (5, 8, 43), and chemical (8, 9, 11, 12, 29, 35, 40, 53) mediators of vasom…”
BackgroundDiNicolantonio, J. J., McCarty, M. F., Assanga, S. B. I., Luján, L. M. L., & O’Keefe, J. H. (2022) · Open Heart · doi:10.1136/openhrt-2021-001801 · Cited by 54 · see citing works
Cardioprotective effects of nitrite during exercise
Calvert, J. W. (2010) · Cardiovascular Research · doi:10.1093/cvr/cvq307 · Cited by 51 · see citing works
“Exercise has been associated with increasing plasma nitrite levels in both rodents and humans.(9,75,76) However, the increased circulating levels of nitrite have traditionally been considered only as an acute marker of NO production and a surrogate for endothelial function.”
BackgroundDzeja, P. P., Bast, P., Pucar, D., Wieringa, B., & Terzic, A. (2007) · Journal of Biological Chemistry · doi:10.1074/jbc.m705268200 · Cited by 50 · see citing works
Vascular remodeling: A redox-modulated mechanism of vessel caliber regulation
Tanaka, L. Y., & Laurindo, F. R. M. (2017) · Free Radical Biology and Medicine · doi:10.1016/j.freeradbiomed.2017.01.025 · Cited by 50 · see citing works
Protein kinase N2 mediates flow-induced eNOS activation and vascular tone regulation
Jin, Y., Chennupati, R., Li, R., Liang, G., Wang, S., & Iring, A. (2021) · Repository of the Academy's Library (Library of the Hungarian Academy of Sciences) · Cited by 49 · see citing works
Wang, C., Li, L., Zhang, Z., Dong, F., Zhu, Y., & Wu, L. (2009) · Journal of Cellular Physiology · doi:10.1002/jcp.21931 · Cited by 47 · see citing works
“This Ad-AMPK-DN inhibits endogenous AMPK activity by replacing the endogenous AMPKa subunit JOURNAL OF CELLULAR PHYSIOLOGY with the inactive form in the abg homoenzyme (Zhang et al., 2006).”
MethodZhu, Z., Fu, C., Li, X., Song, Y., Li, C., Zou, M., Guan, Y., & Zhu, Y. (2011) · PLoS ONE · doi:10.1371/journal.pone.0023554 · Cited by 48 · see citing works
“Ad-AMPK-DN, a recombinant adenovirus expressing a dominant-negative mutant of AMPK, was described previously [15].”
BackgroundDong, Q., Yang, Y., Song, L., Qian, H., & Xu, Z.-M. (2010) · International Journal of Cardiology · doi:10.1016/j.ijcard.2010.08.047 · Cited by 47 · see citing works
Activation of AMP-activated protein kinase by tributyltin induces neuronal cell death
Nakatsu, Y., Kotake, Y., Hino, A., & Ohta, S. (2008) · Toxicology and Applied Pharmacology · doi:10.1016/j.taap.2008.03.021 · Cited by 48 · see citing works
“Involvement of AMPK in nitric oxide synthase (NOS) activation NOS activation is reported to be downstream of AMPK (Zhang et al., 2006).”
BackgroundCheng, C. K., Shang, W., Liu, J., Cheang, W. S., Wang, Y., Li, X., Lau, C., Luo, J., Ng, C., Huang, Y., et al. (2022) · Antioxidants · doi:10.3390/antiox11061137 · Cited by 47 · see citing works
“Additionally, LSS promotes AMPK activation in endothelial cells [54].”
BackgroundRichter, R. P., Ashtekar, A. R., Zheng, L., Pretorius, D., Tripathi, K. K., Sanderson, R. D., Gaggar, A., & Richter, J. R. (2022) · JCI Insight · doi:10.1172/jci.insight.155010 · Cited by 45 · see citing works
“(46) showed that eNOS activity may also be controlled by AMPK signaling.”
BackgroundSu, K., Yu, Y., Hou, H., Zhao, J., Kou, Y. R., Cheng, L., Shyue, S., & Lee, T. (2011) · Journal of Cellular Physiology · doi:10.1002/jcp.23052 · Cited by 45 · see citing works
“Furthermore, arterial AMPK activity and phosphorylation of eNOS were found increased in exercisetraining mice (Zhang et al., 2006; Zhang et al., 2009).”
BackgroundVaanholt, L. M., Jónás, I., Doornbos, M. E., Schubert, K. A., Nyakas, C. J., Garland, T., Visser, G. H., & van Dijk, G. (2008) · International Journal of Obesity · doi:10.1038/ijo.2008.136 · Cited by 44 · see citing works
Lim, J. C. L., Henry, C. J., & Haldar, S. (2016) · Molecular Nutrition & Food Research · doi:10.1002/mnfr.201600121 · Cited by 45 · see citing works
Flow-Mediated Signaling Modulates Endothelial Cell Phenotype
Garin, G., & Berk, B. C. (2006) · Endothelium · doi:10.1080/10623320601061599 · Cited by 44 · see citing works
Anthracycline chemotherapy‐mediated vascular dysfunction as a model of accelerated vascular aging
Clayton, Z. S., Hutton, D. A., Mahoney, S. A., & Seals, D. R. (2021) · Aging and Cancer · doi:10.1002/aac2.12033 · Cited by 43 · see citing works
Baltgalvis, K. A., White, K., Li, W., Claypool, M. D., Lang, W., Alcantara, R. R., Singh, B., Friera, A. M., McLaughlin, J. R., Hansen, D., et al. (2014) · American Journal of Physiology. Heart and Circulatory Physiology · doi:10.1152/ajpheart.00839.2013 · Cited by 41 · see citing works
Targeting the vasculature in cardiometabolic disease
Boutagy, N. E., Singh, A. K., & Sessa, W. C. (2022) · Journal of Clinical Investigation · doi:10.1172/jci148556 · Cited by 43 · see citing works
“Interestingly, in ECs, AMPK activation was demonstrated to activate eNOS (99, 100), thus supporting a bidirectional relationship between energy sensing and NO signaling.”
BackgroundLKB1 Deficiency in Tie2-Cre-expressing Cells Impairs Ischemia-induced Angiogenesis
Ohashi, K., Ouchi, N., Higuchi, A., Shaw, R. J., & Walsh, K. (2010) · Journal of Biological Chemistry · doi:10.1074/jbc.m110.123794 · Cited by 41 · see citing works
- Authors
- Zhang, Y., Lee, T.-S., Kolb, E. M., Sun, K., Lu, X., Sladek, F. M., Kassab, G. S., Garland, T., & Shyy, J. Y.-J.
- Affiliations
- University of California, Riverside (US)
- Venue
- Arteriosclerosis Thrombosis and Vascular Biology
- Published
- 2006-06
- Volume
- 26
- Issue
- 6
- Pages
- 1281-1287
- ISSN
- 1079-5642 (ISSN-L)
- 1524-4636
- OpenAlex
- W2143950284
- Cited by
- 212
- References
- 35
- Citations by year
- 2026: 7 · 2025: 4 · 2024: 6 · 2023: 5 · 2022: 11 · 2021: 15 · 2020: 11 · 2019: 6 · 2018: 8 · 2017: 8 · 2016: 8 · 2015: 11 · 2014: 12 · 2013: 17 · 2012: 8
- FWCI
- 9.28
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Arteriosclerosis Thrombosis and Vascular Biology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Heart, Lung, and Blood Institute (HL-77448, HL55554, R01HL077448)
- National Institute of Diabetes and Digestive and Kidney Diseases (DK053892, R01DK053892)
- Topics
- Health Sciences › Medicine › Physiology › Nitric Oxide and Endothelin Effects
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Metabolism, Diabetes, and Cancer
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
- Keywords
- AMPK, endothelial nitric oxide synthase, shear stress, endothelial cells, LKB1, nitric oxide production, vascular tone, voluntary running, aorta, pulsatility, flow rate
- MeSH terms
- Animals, Aorta, Cattle, Cells, Cultured, Motor Activity, Multienzyme Complexes, Nitric Oxide, Phosphorylation, Protein Kinases, Pulsatile Flow, Stress, Mechanical, Protein Serine-Threonine Kinases, Endothelial Cells, Mice, Nitric Oxide Synthase Type III, AMP-Activated Protein Kinases
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