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Competitive binding of circCCDC6 to microRNA-128-3p activates TXNIP/NLRP3 pathway and promotes cerebral ischemia-reperfusion defects

Wang, ChongShu, Dong, MingMing, Zhang, XiaoYi, Wang, XiaoYu, Zhao, Yan, Cao, Yunpeng·2023·Acta Biochimica Polonica
DOI 10.18388/abp.2020_6552
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Abstract

Objective: Circular RNAs (circRNAs) are enriched in the brain and involved in various central nervous system diseases. The potential role of circCCDC6 in cerebral ischemia-reperfusion defects was partly elucidated in the work. Methods: A middle cerebral artery occlusion/reperfusion (MCAO/R) rat model and an oxygen-glucose deprivation and re-oxygenation (OGD/R)-treated SH-SY5Y cell model were constructed. CircCCDC6 expression in the two models was examined, and circCCDC6-involved mechanisms in neuronal pyroptosis and inflammation were analyzed through loss- and gain-of-function assays. Results: MCAO/R rat brain tissues and OGD/R-treated SH-SY5Y cells exhibited upregulated circCCDC6. Silencing circCCDC6 attenuated neuronal pyroptosis and inflammation in the brain tissue of MCAO/R rats. Overexpressing circCCDC6 or inhibiting miR-128-3p stimulated OGD/R-induced pyroptosis and inflammation in SH-SY5Y cells, while upregulating miR-128-3p attenuated OGD/R injury. CircCCDC6 silencing-induced effects on SH-SY5Y cells were antagonized by TXNIP overexpression. Conclusion: Mechanistically, circCCDC6 mediates miR-128-3p and activates TXNIP/NLRP3, thereby promoting OGD/R-induced neuronal pyroptosis and inflammation. CircCCDC6 may provide a new strategy for the treatment of MCAO/R.

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Figure 1 Effects of injection of shRNA-circCCDC6 lentiviral vector on biochemical indexes and pathological tissue damage of MCAO/R rat brain.
Figure 1 Effects of injection of shRNA-circCCDC6 lentiviral vector on biochemical indexes and pathological tissue damage of MCAO/R rat brain.CC-BY · From the PDF
Figure 2 Effect of transfection of pcDNA 3.1-circCCDC6 on the biological behavior and biochemical indexes of OGD/R-treated SH-
Figure 2 Effect of transfection of pcDNA 3.1-circCCDC6 on the biological behavior and biochemical indexes of OGD/R-treated SH-CC-BY · From the PDF
Figure 3 Binding relation of circCCDC6 and miR-128-3p. Potential binding sites of miR-128-3p and circCCDC6 ( A ), miR-128-3p expression in in vivo and in vitro models ( B ), verification of targeting relationship between circCCDC6 and miR-128-3p ( C-D ), effects of low or overexpression of circCCDC6 on miR-128-3p expression in SHSY5Y cells ( E ). data were reported as mean ± S.D. (N=3).
Figure 3 Binding relation of circCCDC6 and miR-128-3p. Potential binding sites of miR-128-3p and circCCDC6 ( A ), miR-128-3p expression in in vivo and in vitro models ( B ), verification of targeting relationship between circCCDC6 and miR-128-3p ( C-D ), effects of low or overexpression of circCCDC6 on miR-128-3p expression in SHSY5Y cells ( E ). data were reported as mean ± S.D. (N=3).CC-BY · From the PDF
Figure 4 Effects of transfection of miR-128-3p mimic/inhibitor on the biological behavior and biochemical indexes of SH-SY5Y cells treated with OGD/R. miR-128-3p expression ( A ), cell viability ( B ), toxicity ( C ), caspase-1 positive cells ( D ), IL-1β and IL-1β contents ( E ), protein expression of IL-1β, IL-18, cleaved caspase-1, ASC and NLRP3 ( F ); data were reported as mean ± S.D. (N=3).
Figure 4 Effects of transfection of miR-128-3p mimic/inhibitor on the biological behavior and biochemical indexes of SH-SY5Y cells treated with OGD/R. miR-128-3p expression ( A ), cell viability ( B ), toxicity ( C ), caspase-1 positive cells ( D ), IL-1β and IL-1β contents ( E ), protein expression of IL-1β, IL-18, cleaved caspase-1, ASC and NLRP3 ( F ); data were reported as mean ± S.D. (N=3).CC-BY · From the PDF
Figure 5 Targeting relation between miR-128-3p and TXNIP. TXNIP expression in in vivo and in vitro models ( A-B ), effect of miR-1283p mimic/inhibitor on TXNIP protein expression in SH-SY5Y cells ( C ), potential binding sites of miR-128-3p and TXNIP ( D ), verification of the targeting relationship between miR-128-3p and TXNIP ( E-F ); data were reported as mean ± S.D. (N=3).
Figure 5 Targeting relation between miR-128-3p and TXNIP. TXNIP expression in in vivo and in vitro models ( A-B ), effect of miR-1283p mimic/inhibitor on TXNIP protein expression in SH-SY5Y cells ( C ), potential binding sites of miR-128-3p and TXNIP ( D ), verification of the targeting relationship between miR-128-3p and TXNIP ( E-F ); data were reported as mean ± S.D. (N=3).CC-BY · From the PDF
Figure 6 Effects of transfection of si-circCCDC6 and pcDNA 3.1-TXNIP on the biological behavior and biochemical indicators of OGD/R-treated SH-SY5Y cells.
Figure 6 Effects of transfection of si-circCCDC6 and pcDNA 3.1-TXNIP on the biological behavior and biochemical indicators of OGD/R-treated SH-SY5Y cells.CC-BY · From the PDF

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Metadata

Authors
Wang, C., Dong, M., Zhang, X., Wang, X., Zhao, Y., & Cao, Y.
ORCID
Wang, ChongShu
Venue
Acta Biochimica Polonica
Publisher
Frontiers Media SA
Published
2023-11-07
ISSN
  • 1734-154X
  • 0001-527X
DOI
10.18388/abp.2020_6552
Cited by
2
References
0
Licence
creativecommons.org/licenses/by/4.0 (unspecified)

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