【摘要】 目的 考察佛手苷内酯(bergapten,BG)是否通过下调miR-503表达抑制高糖(high glucose,HG)诱导的肾小管上皮细胞炎性因子表达和细胞凋亡。方法 将肾小管上皮细胞分为正常对照(Con)组、高糖(HG)组、高糖+BG低、中、高浓度(HG+BG-L、-M、-H)组、HG+inhibitor-NC组、HG+miR-503 inhibitor组、HG+BG+miR-NC组、HG+BG+miR-503组。利用酶联免疫吸附测定法(ELISA)测定炎性因子肿瘤坏死因子-α (TNF-α)、白细胞介素-6(IL-6)表达,流式细胞术测定细胞凋亡,Western印迹测定凋亡蛋白裂解天冬氨酸特异性半胱氨酸蛋白酶(cleaved-caspase) 3、cleaved-caspase 9表达,qRT-PCR测定miR-503表达。结果 HG组肾小管上皮细胞的TNF-α、IL-6表达、凋亡率、cleaved-caspase 3、cleaved-caspase 9蛋白、miR-503表达量均比Con组增加(P_均<0.05)。HG+BG-L组、HG+BG-M组、HG+BG-H组肾小管上皮细胞的TNF-α、IL-6表达、凋亡率、cleaved-caspase 3、cleaved-caspase 9蛋白、miR-503表达量均比HG组减少(P_均<0.05),且BG的抑制作用呈现浓度依赖性。干扰miR-503表达后,HG+miR-503 inhibitor组肾小管上皮细胞的miR-503表达量、TNF-α、IL-6表达、凋亡率、cleaved-caspase 3蛋白、cleaved-caspase 9蛋白均比HG+inhibitor-NC组降低(P_均<0.05)。HG+BG+miR-503组肾小管上皮细胞的miR-503表达量、TNF-α、IL-6表达、凋亡率、cleaved-caspase 3蛋白、cleaved-caspase 9蛋白表达量均高于HG+BG+miR-NC组(P_均<0.05)。结论 BG通过下调miR-503的表达,以浓度依赖方式抑制高糖诱导的肾小管上皮细胞炎性因子表达和细胞凋亡。更多还原

【Abstract】 Objective To investigate the effect of bergapten(BG) on the inhibition of high glucose(HG) induced inflammatory factors expression and apoptosis in renal tubular epithelial cells by down-regulation of miR-503.Methods The renal tubular epithelial cells were divided into control(Con) group,high glucose(HG) group,high glucose+BG low-,medium-and high concentration(HG+BG-L,-M,-H) group,HG+inhibitor-NC group,HG+miR-503 inhibitor group,HG+BG+miR-NC group,HG+BG+miR-503 group.Enzyme-linked immunosorbent assay(ELISA) was used to measure the levels of inflammatory factors tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6),flow cytometry was used to monitor the cell apoptosis,and Western blotting was used to determine the expression levels of apoptotic proteins cleaved-caspase 3 and cleaved-caspase 9,and the expression level of miR-503 was determined by qRT-PCR.Results The apoptosis rate,the levels of TNF-α,IL-6,and the expression levels of cleaved-caspase 3,cleaved-caspase 9 protein and miR-503 in renal tubular epithelial cells in the HG group were all higher than those in the Con group(all P<0.05).The apoptosis rate,the levels of TNF-α,IL-6,and the expression levels of cleaved-caspase 3,cleaved-caspase 9 protein,miR-503 of renal tubular epithelial cells in the HG+BG-L group,the HG+BG-M group,and the HG+BG-H group were all lower than those in the HG group(all P<0.05),and the inhibitory effects of BG were concentration-dependent.After interfering with miR-503,the expression levels of miR-503,cleaved-caspase 3 and cleaved-caspase 9,the levels of TNF-α,IL-6,and the apoptosis rate in renal tubular epithelial cells in the HG+miR-503 inhibitor group were lower than those in the HG+inhibitor-NC group(all P <0.05).The expression levels of miR-503,cleaved-caspase 3 and cleaved-caspase 9,the levels of TNF-α,IL-6,and the apoptosis rate in renal tubular epithelial cells in the HG+BG+miR-503 group were higher than those in the HG+BG+miR-NC group(all P<0.05).Conclusion BG inhibits the high glucose induced expression of inflammatory factors and apoptosis of renal tubular epithelial cells in a concentration-dependent manner by down-regulation of the miR-503 expression.更多还原