[关键词]
[摘要]
目的 探究小檗碱对高糖诱导的足细胞损伤的影响及其对miR-1290的调控作用。方法 采用高糖诱导小鼠肾足细胞MPC5建立细胞损伤模型,给予不同浓度(1.0、2.5、5.0 μmol/L)小檗碱进行干预,采用流式细胞术检测细胞凋亡情况;采用Western blotting法检测细胞B淋巴细胞瘤2(B-cell lymphoma 2,Bcl-2)和Bcl-2相关X蛋白(Bcl-2-associated X protein,Bax)蛋白表达情况;采用ELISA法检测细胞上清液中白细胞介素-6(interleukin-6,IL-6)、IL-1β、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)水平;采用qRT-PCR法检测miR-1290 mRNA表达情况。将阴性对照(miR-NC)和miR-1290模拟物(miR-1290 mimics)分别转染至MPC5细胞,给予5 μmol/L小檗碱进行干预,考察miR-1290对小檗碱抑制高糖诱导的细胞凋亡及炎性因子分泌的影响。结果 与对照组比较,模型组细胞凋亡率显著升高(P<0.05),Bcl-2蛋白表达水平显著降低(P<0.05),Bax蛋白表达水平显著升高(P<0.05),上清液中IL-6、IL-1β和TNF-α水平显著升高(P<0.05),miR-1290 mRNA表达水平显著升高(P<0.05)。与模型组比较,小檗碱组细胞凋亡率显著降低(P<0.05),Bcl-2蛋白表达水平显著升高(P<0.05),Bax蛋白表达水平显著降低(P<0.05),上清液中IL-6、IL-1β和TNF-α水平显著降低(P<0.05),miR-1290 mRNA表达水平显著降低(P<0.05)。与小檗碱+miR-NC组比较,小檗碱+miR-1290 mimics组细胞凋亡率显著升高(P<0.05),Bcl-2蛋白表达水平显著降低(P<0.05),Bax蛋白表达水平显著升高(P<0.05),上清液中IL-6、IL-1β和TNF-α水平显著升高(P<0.05)。结论 小檗碱能够通过下调miR-1290表达抑制高糖诱导的足细胞凋亡及炎性反应,从而减轻细胞损伤。
[Key word]
[Abstract]
Objective To explore the effect of berberine on podocyte damage induced by high glucose and its regulatory effect on miR-1290. Methods MPC5 cells was induced by high glucose to establish a cell injury model, and different concentrations (1.0, 2.5, 5.0 μmol/L) of berberine were given for intervention. The apoptosis was detected by flow cytometry; Western blotting was used to detect B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X protein (Bax) protein expressions; ELISA method was used to detect interleukin-6 (IL-6), IL-1β, and tumor necrosis factor-α (TNF-α) levels in supernatant; qRT-PCR method was used to detect miR-1290 mRNA expression. Negative control (miR-NC) and miR-1290 mimics were respectively transfected into MPC5 cells, and 5 μmol/L berberine was given for intervention to investigate the effect of miR-1290 on berberine in inhibiting apoptosis and inflammatory factors secretion in MPC5 cells induced by high glucose. Results Compared with control group, apoptosis rate of MPC5 cells in model group was significantly increased (P<0.05), Bcl-2 protein expression level was significantly reduced (P<0.05), Bax protein expression level was significantly increased (P<0.05), IL-6, IL-1β and TNF-α levels in supernatant were significantly increased (P<0.05), and miR-1290 mRNA expression was significantly increased (P<0.05). Compared with model group, apoptosis rate of MPC5 cells in berberine group was significantly reduced (P<0.05), Bcl-2 protein expression level was significantly increased (P<0.05), Bax protein expression level was significantly reduced (P<0.05), IL-6, IL-1β and TNF-α levels in supernatant were significantly reduced (P<0.05), and miR-1290 mRNA expression levels were significantly reduced (P<0.05). Compared with berberine + miR-NC group, apoptosis rate of MPC5 cells in berberine + miR-1290 mimics group was significantly increased (P<0.05), Bcl-2 protein expression level was significantly reduced (P<0.05), Bax protein expression level was significantly increased (P<0.05), IL-6, IL-1β and TNF-α levels in supernatant were significantly increased (P<0.05). Conclusion Berberine can inhibit podocyte apoptosis and inflammatory response induced by high glucose through down-regulating miR-1290 expression, thereby reducing cell damage.
[中图分类号]
R285.5
[基金项目]
国家重点研发计划子课题(2017YFC0113901)