恩格列净通过miR-497-3p/GPLD1通路调控高糖诱人肾小球微血管内皮细胞增殖凋亡的机制研究

doi:10.3969/j.issn.1006-6233.2022.02.04

Abstract
Figure/Table
References
Related Citation (15)

Download: PDF (2023 KB) HTML (1 KB)
Export: BibTeX | EndNote (RIS)

Abstract Objective: To explore the mechanism of empagliflozin (EMPA) regulating the proliferation and apoptosis of human glomerular microvascular endothelial cells (HRGEC). Methods: HRGEC was induced by 30 mmoL/L glucose to establish the renal injury cell model in high glucose environment. The damaged cells were treated with englitazin (250, 500, 1000nmoL/L). HRGEC cells were transfected with miR-NC group (transfected miR-NC), miR-497-3p group (transfected miR-497-3p), si-NC group (transfected si-NC), si-fish glycosylphosphatidylinositol specific phospholipase D1 (GPLD1) group (transfected si-GPLD1), EMPA + miR-497-3p + pcDNA group (co-transfected miR-497-3p and pcDNA), EMPA + miR-497-3p + pcDNA-GPLD1 group (co-transfected miR-497-3p and pcDNA-GPLD1), were all treated with 30mmoL/L glucose or 500 nmol / L empagliflozin. The cell proliferation rate and apoptosis rate were detected by cell counting kit (CCK-8) method and flow cytometry. The expressions of GPLD1, proliferating cell nuclear antigen (PCNA), cell cycle dependent protein kinase inhibitor 1a (p21), Caspase-3 and Caspase-9 were detected by western blotting (WB) assay. The binding ability of miR-497-3p to GPLD1 was detected by dual luciferase assay and RNA binding protein immunoprecipitation assay (RIP). Results: Compared with NG group, the proliferation rate significantly decreased in HG group, the apoptosis rate significantly increased, and the expression of miR-497-3p significantly decreased (P<0.05). Compared with HG group, the cell proliferation rate, apoptosis rate and the expression of miR-497-3p were significantly increased in empagliflozin (250, 500 and 1000nmoL/L) group (P<0.05). The optimal concentration of empagliflozin was 500 nmol/L. MiR-497-3p significantly inhibited the fluorescence activity of wild-type GPLD1 cells, promoted the expression of Ago2-GPLD1, and negatively regulated the protein expression of GPLD1. Overexpression of miR-497-3p and knockdown of GPLD1 had similar effects on promoting the proliferation and inhibiting apoptosis, up-regulating PCNA protein and down-regulating p21, caspase-3 and caspase-9 protein of HRGEC cells induced by high glucose. Overexpression of GPLD1 significantly weakened the regulatory effects on the proliferation and apoptosis of empagliflozin and overexpression of miR-497-3p in HRGEC cells induced by high glucose. Conclusion: Empagliflozin could promote the proliferation and inhibit apoptosis of HRGEC cells induced by high glucose, and its mechanism is related to the regulation of miR-497-3p / GPLD1 signal pathway.

Key words： Empagliflozin Diabetes MiR-497-3p GPLD1

	Service

	E-mail this article
	Add to my bookshelf
	Add to citation manager
	E-mail Alert
	RSS
	Articles by authors

Cite this article:

URL:

http://www.hbyxzzs.cn/EN/10.3969/j.issn.1006-6233.2022.02.04 OR http://www.hbyxzzs.cn/EN/Y2022/V28/I2/194

[1] Frampton JE.Empagliflozin: a review in type 2 diabetes[J].Drugs,2018,78(10):1037~1048.
[2] Levine MJ.Empagliflozin for type 2 diabetes mellitus:an overview of phase 3 clinical trials[J].Curr Diabetes Rev,2017,13(4):405~423.
[3] Zannad F,Ferreira JP,Pocock SJ,et al.Cardiac and kidney benefits of empagliflozin in heart failure across the spectrum of kidney function:insights from EMPEROR-reduced[J].Circulation,2021,143(4):310~321.
[4] Vishnoi A,Rani S.MiRNA biogenesis and regulation of diseases:an overview[J].Methods Mol Biol,2017,1509(11):1~10.
[5] Li Y,Yuan X,Shi Z,et al.LncRNA XIST serves as a diagnostic biomarker in gestational diabetes mellitus and its regulatory effect on trophoblast cell via miR-497-5p/FOXO1 axis[J].Cardiovasc Diagn Ther,2021,11(3):716~725.
[6] Anker SD,Butler J,Filippatos G,et al.Effect of empagliflozin on cardiovascular and renal outcomes in patients with heart failure by baseline diabetes status:results from the EMPEROR-reduced Trial[J].Circulation,2021,143(4):337~349.
[7] Packer M,Anker SD,Butler J,et al.Cardiovascular and renal outcomes with empagliflozin in heart failure[J].N Engl Med,2020,383(15):1413~1424.
[8] Gonzalez DE,Foresto RD,Ribeiro AB.SGLT-2 inhibitors in diabetes:a focus on renoprotection[J].Rev Assoc Med Bras (1992),2020,66(Suppl 1):s17~s24.
[9] Lee YH,Kim SH,Kang JM,et al.Empagliflozin attenuates diabetic tubulopathy by improving mitochondrial fragmentation and autophagy[J].Am Physiol Renal Physiol,2019,317(4):767~780.
[10] Nitin AD,Andrea JC,Anthony B,et al.Empagliflozin reduces high glucose-induced oxidative stress and miR-21-dependent TRAF3IP2 induction and RECK suppression,and inhibits human renal proximal tubular epithelial cell migration and epithelial-to-mesenchymal transition[J].Cell Signal,2020,68(4):109506.
[11] Hathaway QA,Pinti MV,Durr AJ,et al.Regulating microRNA expression:at the heart of diabetes mellitus and the mitochondrion[J].Am Physiol Heart Circ Physiol,2018,314(2):293~310.
[12] Ban E,Jeong S,Park M,et al.Accelerated wound healing in diabetic mice by miRNA-497 and its anti-inflammatory activity[J].Biomed Pharmacother,2020,121(1):109613.
[13] Wang J,Zhao SM.LncRNA-antisense non-coding RNA in the INK4 locus promotes pyroptosis via miR-497/thioredoxin-interacting protein axis in diabetic nephropathy[J].Life Sci,2021,264(1):118728.
[14] Abdolmaleki F,Heidarianpour A.Endurance exercise training restores diabetes-induced alteration in circulating glycosylphosphatidylinositol-specific phospholipase D levels in rats[J].Diabetol Metab Syndr,2020,12(5):43.
[15] Zhang K,Zhu H,Wang L,et al.Serum glypican4 and glycosylphosphatidylinositol-specific phospholipase D levels are associated with adipose tissue insulin resistance in obese subjects with different glucose metabolism status[J].Endocrinol Invest,2021,44(4):781~790.