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目的探讨活性维生素D3(Vit D3)对高糖诱导的人肾小管上皮细胞(HK-2)中α-平滑肌肌动蛋白(α-SMA)和纤维连接蛋白(FN)表达的影响。方法培养人肾小管上皮细胞并分为正常组(N组,含5.5 mol.L-1D-葡萄糖)、高糖组(M组,含25 mol.L-1D-葡萄糖)、高糖+低浓度活性Vit D3组(LD组,含10-8mol.L-1活性Vit D3)和高糖+高浓度活性Vit D3组(HD组,含10-6mol.L-1活性维生素D3),细胞免疫组化方法测定各组细胞的α-SMA和FN的蛋白表达,RT-PCR半定量方法测定各组细胞的α-SMA和FN的mRNA表达。结果与N组比较,M组、LD组和HD组α-SMA、FN蛋白表达的水平和mRNA表达的水平明显升高(P<0.05),与M组比较,LD组和HD组α-SMA、FN蛋白表达的水平和mRNA表达的水平明显降低(P<0.05),其中,与LD组比较,HD组水平最低(P<0.05)。结论活性Vit D3可下调人肾小管上皮细胞α-SMA和FN的表达,从而抑制肾小管上皮细胞转分化,对糖尿病肾病有一定防治作用,且存在剂量依赖关系。
Objective To investigate the effects of active vitamin D3 (Vit D3) on the expression of α-smooth muscle actin (α-SMA) and fibronectin (FN) in high glucose-induced human renal tubular epithelial cells (HK- Methods Human renal tubular epithelial cells were cultured and divided into normal group (N group, containing 5.5 mol.L-1D-glucose), high glucose group (M group, containing 25 mol.L-1D-glucose), high glucose + low concentration Active Vit D3 group (LD group, containing 10-8mol.L-1 active Vit D3) and high glucose + high concentration Vit D3 group (HD group, containing 10-6mol.L-1 active vitamin D3), cellular immunity group Methods The α-SMA and FN protein expression in each group of cells was determined. The mRNA expression of α-SMA and FN in each group was determined by semi-quantitative RT-PCR. Results Compared with the N group, the expression of α-SMA, FN and the expression of α-SMA were significantly increased in M group, LD group and HD group (P <0.05) (P <0.05). The level of FN protein expression and mRNA expression were significantly lower in HD group than in LD group (P <0.05). Conclusion The active Vit D3 can down-regulate the expression of α-SMA and FN in human renal tubular epithelial cells and thus inhibit the renal tubular epithelial cell transdifferentiation, which may prevent and treat diabetic nephropathy in a dose-dependent manner.