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目的探讨α-硫辛酸(ALA)对高糖诱导的肾小管上皮细胞转分化的影响。方法体外培养人肾小管上皮细胞株(HKC)分别予正常糖(NG组)、高糖(HG组)、高糖+ALA(100、200、400μmol/L)组或高糖+甘露醇(M组)干预48h;观察HKC细胞形态改变,RT-PCR检测结缔组织生长因子(CTGF)和α-平滑肌肌动蛋白(α-SMA)mRNA的表达,免疫荧光染色检测α-SMA蛋白表达。结果高糖刺激使HKC细胞由铺路石样变为梭形长条状;ALA干预可部分逆转高糖对HKC细胞形态的影响。HG组干预HKC细胞的CTGF及α-SMA mRNA表达水平较NG组增强,高糖+ALA组表达均较HG组减弱,且呈浓度依赖性(P<0.01);HG组α-SMA表达增强,NG组表达微弱,高糖+ALA组表达渐次减弱,差异均有统计学意义(P<0.01);M组上述指标与NG组比较差异均无统计学意义。结论 ALA可阻断高糖诱导的肾小管上皮细胞转分化,对糖尿病慢性肾脏疾病(CKD)的防治可能具有一定作用。
Objective To investigate the effects of α-lipoic acid (ALA) on the transdifferentiation of renal tubular epithelial cells induced by high glucose. Methods Human renal tubular epithelial cell line (HKC) was cultured with normal glucose (NG), high glucose (HG), high glucose + ALA (100,200,400μmol / L) Group) for 48h. The morphological changes of HKC cells were observed. The expression of CTGF and α-SMA mRNA was detected by RT-PCR. The expression of α-SMA protein was detected by immunofluorescence staining. Results High glucose stimulated HKC cells to change from paving stones to fusiform strips. ALA intervention partially reversed the effect of high glucose on the morphology of HKC cells. The expression of CTGF and α-SMA mRNA in HG group was significantly higher than that in NG group. The expression of α-SMA in HG group was weaker than that in HG group and in a concentration-dependent manner (P <0.01) The expression of NG in group A was weaker than that in group A, and the expression in high glucose + ALA group was gradually weakened (P <0.01). There was no significant difference in the above indexes between M group and NG group. Conclusion ALA can block high glucose-induced renal tubular epithelial cell transdifferentiation, which may play a role in the prevention and treatment of chronic kidney disease (CKD) in diabetes.