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扬州大学附属医院内分泌科,扬州,225000
纸质出版日期:2022
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孙枫. miR-132-5p靶向S100A9介导高糖诱导的肾小球系膜细胞凋亡及炎症反应[J]. 重庆医科大学学报, 2022,47(6):629-634.
Sun Feng. miR-132-5p mediates apoptosis and inflammatory of high glucose-induced mesangial cells by targeting S100A9[J]. Journal of chongqing medical university, 2022, 47(6): 629-634.
目的: 探讨微小RNA-132-5p(microRNA-132-5p,miR-132-5p)靶向S100钙结合蛋白A9(S100 calcium binding protein A9,S100A9)对高糖诱导的肾小球系膜细胞凋亡及炎症反应的影响。方法: 人肾小球系膜细胞用DMEM正常糖培养基培养作为对照组,用DMEM高糖培养基培养作为高糖组;miR-NC、miR-132-5p、anti-miR-NC、anti-miR-132-5p转染至正常培养的肾小球系膜细胞中,记为miR-NC组、miR-132-5p组、anti-miR-NC组、anti-miR-132-5p组;将miR-NC、miR-132-5p、si-NC、si-S100A9转染至肾小球系膜细胞中再用高糖培养基培养,记为高糖+miR-NC组、高糖+miR-132-5p组、高糖+si-NC组、高糖+si-S100A9组;将miR-132-5p分别与pcDNA、pcDNA-S100A9转染至肾小球系膜细胞中再用高糖培养基培养,记为高糖+miR-132-5p+pcDNA组、高糖+miR-132-5p+pcDNA-S100A9组。实时荧光定量PCR(real-time quantitative PCR,RT-qPCR)检测miR-132-5p和S100A9 mRNA表达水平;Western blot检测S100A9、Bcl-2相关X蛋白(Bcl-2 associated X,Bax)、B细胞淋巴瘤/白血病-2蛋白(B cell lymphoma/leukemia-2,Bcl-2)表达;双荧光素酶报告实验验证miR-132-5p和S100A9的靶向关系;流式细胞术检测细胞凋亡;酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)检测肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白介素(interleukin,IL)-8、IL-6水平。结果: 高糖诱导的肾小球系膜细胞中miR-132-5p表达水平降低,S100A9表达水平升高(P<0.05)。miR-132-5p靶向调控S100A9,过表达miR-132-5p或抑制S100A9表达,高糖诱导的肾小球系膜细胞中细胞凋亡率降低,Bax表达水平降低,Bcl-2表达水平升高,TNF-α、IL-8、IL-6水平降低(P<0.05)。S100A9过表达逆转miR-132-5p过表达对高糖诱导的肾小球系膜细胞凋亡和炎症反应的抑制作用。结论: 过表达miR-132-5p通过靶向下调S100A9抑制高糖诱导的肾小球系膜细胞凋亡和炎症反应。
Objective: To investigate the effect of microRNA-132-5p (miR-132-5p) targeting S100 calcium-binding protein A9 (S100A9) on the apoptosis and inflammatory of mesangial cells induced by high glucose. Methods: Human mesangial cells were cultured in DMEM normal glucose medium as control group
and cultured in DMEM high glucose medium as high glucose group.miR-NC
miR-132-5p
anti-miR-NC
anti-miR-132-5p were transfected into normal cultured mesangial cells and recorded as miR-NC group
miR-132-5p group
anti-miR-NC group
anti-miR-132-5p group; miR-NC
miR-132-5p
si-NC
si-S100A9 were transfected into mesangial cells and cultured in a high glucose medium
and recorded as high glucose+miR-NC group
high glucose+miR-132-5p group
high glucose+si-NC group
high glucose+si-S100A9 group; miR-132-5p was transfected with pcDNA and pcDNA-S100A9 into mesangial cells and cultured with high glucose medium
and recorded as high glucose+miR-132-5p pcDNA group
high glucose+miR-132-5p+pcDNA-S100A9 group. Real-time quantitative PCR (RT-qPCR) was used to detect miR-132-5p and S100A9 mRNA expressions. Western blot was used to detect S100A9
Bcl-2 associated X protein (Bax)
and B cells lymphoma/leu-kemia-2 protein (Bcl-2) expression; dual luciferase assay was used to verify the targeting relationship between miR-132-5p and S100A9; flow cytometry was used to detect apoptosis; enzyme-linked immu-nosorbent assay (ELISA) detected the levels of tumor necrosis factor-α (TNF-α)
interleukin (IL) -8 and IL-6. Results: The expression of miR-132-5p in mesangial cells induced by high glucose was decreased
and the expression of S100A9 was increased (P<0.05).miR-132-5p targetedS100A9
overexpressedmiR-132-5porinhibited S100A9 expression. Apoptosis was reduced in high glucose-induced mesangial cells
Bax expression was reduced
Bcl-2 expression was increased
and TNF-α
IL-8
IL-6 levels were decreased (P<0.05). S100A9 overexpression reversed the inhibitory effect of miR-132-5p overexpression on apoptosis and inflammatory of mesangial cells induced by high glucose. Conclusion: Overexpression of miR-132-5p can inhibit apoptosis and inflammatory of high glucose-induced mesangial cells by down-regulating S100A9.
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