Abstract：Objective: To observe the effects of miR-93-5p and dual-specificity phosphatase 8 (DUSP8) on high glucose-induced HPC cells, and explore its mechanism. Methods: HPC cells were treated with 25mM high glucose for 6, 12, and 24 hours. The optimal treatment time was 12h by the secretion of the inflammatory factors interleukin-6 (IL-6) and Tumor Necrosis Factor-alpha (TNF-α) in the cells, apoptosis rate and apoptosis-related protein split Cleaved caspase-3 (Cleaved caspase-3), Bcl-2 associated X protein (Bax), B-cell lymphoma-2(Bcl-2). The miR-con group (transfected miR-con), miR-93-5p group (transfected miR-93-5p mimics), anti-miR-con group (transfected anti-miR-con), anti-miR- 93-5p group (transfected anti-miR-93-5p mimics), high glucose (HG)+si-con group (transfected si-con), HG+si-DUSP8 group (transfected si-DUSP8), HG+miR-93-5p +pcDNA group (co-transfected miR-93-5p mimics and pcDNA), HG+miR-93-5p+ pcDNA-DUSP8 group (co-transfected miR-93-5p mimics and pcDNA-DUSP8) were all transfected to HPC cells with liposome methods . Fluorescence quantitative polymerase chain reaction (qRT-PCR), Western blotting (WB), Enzyme linked immunosorbent assay (ELISA), flow cytometry were used to detect the expression of miR-93-5p, DUSP8, cell secretion of IL-6, TNF-α , and protein expression of DUSP8, Cleaved caspase-3, Bax, Bcl-2, and cell apoptosis rate; Dual luciferase reporter experiment detects the fluorescence activity of cells. Results: Compared with the low glucose (NG) group, the expression of miR-93-5p was significantly decreased, the expression of DUSP8 was substantially increased, the levels of IL-6 and TNF-α were substantially increased, protein expression of Cleaved caspase-3, Bcl-2 were significantly decreased, Bax protein expression was significantly increased, the cell apoptosis rate was markedly increased in the HG (6, 12, 24 h) group. The optimal treatment time was 12 h. Overexpression miR-93-5p depressed the secretion of IL-6, TNF-α, apoptosis rate, and facilitated proteins of Cleaved caspase-3 and Bcl-2, and suppressed Bax protein in HPC cells induced by high glucose. MiR-93-5p inhibited the fluorescent activity of wild-type DUSP8 cells. Silencing DUSP8 had a protective effect similar to that of overexpression miR-93-5p, and overexpression DUSP8 reversed the effects of overexpression miR-93-5p on the inhibition of IL-6 and TNF- α secretion and apoptosis in injured podocytes. Conclusion: MiR-93-5p could inhibit the secretion and apoptosis of inflammatory factors in podocytes induced by high glucose, and its mechanism is related to miR-93-5p targeting DUSP8.
顾勇, 杨艳, 李娜, 王艳, 张姣, 杜婷, 范小平. miR-93-5p靶向DUSP8基因对糖尿病肾病足细胞损伤的影响[J]. 河北医学, 2022, 28(1): 21-27.
GU Yong, YANG Yan, LI Na, et al. The Effects of MiR-93-5p on Podocyte Damage in Diabetic Nephropathy by Targeting DUSP8 Gene. HeBei Med, 2022, 28(1): 21-27.
[1] Nagib AM,Elsayed Matter Y,Gheith OA,et al.Diabetic nephropathy following posttransplant diabetes mellitus[J].Exp Clin Transplant,2019,17(2):138~146. [2] Marathias KP,Lambadiari VA,Markakis KP,et al.Competing effects of renin angiotensin system blockade and sodium-glucose cotransporter-2 inhibitors on erythropoietin secretion in diabetes[J].Am Nephrol,2020,51(5):349~356. [3] Nistor I,De Sutter J,Drechsler C,et al.Effect of renin-angiotensin-aldosterone system blockade inadults with diabetes mellitus and advanced chronic kidney disease not on dialysis:a systematic review and meta-analysis[J].Nephrol Dial Transplant,2018,33(1):12~22. [4] Vishnoi A,Rani S.MiRNA biogenesis and regulation of diseases:An overview[J].Methods Mol Biol,2017,1509(11):1~10. [5] Vasu S,Kumano K,Darden CM,et al.MicroRNA signatures as future biomarkers for diagnosis of diabetes states[J].Cells,2019,8(12):1533~1541. [6] Li JZ,Zhao Q,Jin XH,et al.Silencing of LncRNA PVT1 inhibits the proliferation,migration and fibrosis of high glucose-induced mouse mesangial cells via targeting microRNA-93-5p[J].Biosci Rep,2020,40(5):4427~4443. [7] Zhu MC,Ye MS,Wang J,et al.Construction of potential miRNA-mRNA regulatory network in COPD plasma by bioinformatics analysis[J].Int Chron Obstruct Pulmon Dis,2020,15(9):2135~2145. [8] Yang JD,Shen Y,Yang X,et al.Silencing of long noncoding RNA XIST protects against renal interstitial fibrosis in diabetic nephropathy via microRNA-93-5p-mediated inhibition of CDKN1A[J].American Journal of Physiology,2019,317(5):F1350~F1358. [9] Zhou M,Hou YL,Wu J,et al.miR-93-5p promotes insulin resistance to regulate type 2 diabetes progression in HepG2 cells by targeting HGF[J].Mol Med Rep,2021,23(5):329~362. [10] Ding T,Zhou Y,Long RY,et al.DUSP8 phosphatase:structure,functions,expression regulation and the role in human diseases[J].Cell Biosci,2019,9(8):70~83. [11] Schriever SC,Kabra DG,Pfuhlmann K,et al.Type 2 diabetes risk gene DUSP8 regulates hypothalamic Jnk signaling and insulin sensitivity[J].Clin Invest,2020,130(11):6093~6108. [12] Peter Baumann,Sonja C,Schriever,et al.Diabetes type 2 risk gene Dusp8 is associated with altered sucrose reward behavior in mice and humans[J].Brain Behav,2021,11(1):1928~1939.
2022, Vol. 28 
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