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目的:探讨大黄酸对人肾小管上皮细胞(HK-2)血小板反应蛋白1(TSP-1)和转化生长因子β1(TGF-β1)表达的影响。方法:体外培养HK-2细胞,观察AngⅡ(1×10-7mol/L)诱导下低、中和高浓度的大黄酸(10μg/ml、20μg/ml和40μg/ml)对HK-2TSP-1、TGF-β1的mRNA和蛋白表达的影响。采用Real Time PCR检测细胞TSP-1、TGF-β1mRNA表达,采用Westernb lot检测TSP-1、TGF-β1蛋白表达。培养上清液中TGF-β1和活性TGF-β1检测采用ELISA法。结果:与空白对照组比较,AngⅡ(1×10-7mol/L)明显上调了HK-2TSP-1的mRNA表达(3.84±0.48vs1.32±0.19;P<0.05)、伴有TGF-β1的mRNA表达增加(3.68±0.46vs1.18±0.13;P<0.05)。同时AngⅡ(1×10-7mol/L)也上调TSP-1蛋白表达(0.5246±0.0936 vs 0.1015±0.0768;P<0.01),伴有TGF-β1蛋白表达的增加(0.5948±0.0624 vs 0.1264±0.0811;P<0.01)。与AngⅡ阳性对照组比较,中浓度和高浓度大黄酸明显抑制了AngⅡ(1×10-7mol/L)诱导的TSP-1、TGF-β1的mRNA(P<0.05,P<0.01)和蛋白表达(P<0.05,P<0.01)。中浓度大黄酸干预组总的TGF-β1含量和活性TGF-β1含量均减少,(P<0.05);高浓度大黄酸干预组总的TGF-β1含量和活性TGF-β1含量均明显减少(P<0.01)。结论:大黄酸能抑制人肾小管上皮细胞TSP-1、TGF-β1的mRNA和蛋白表达,抑制TGF-β分泌。
Objective: To investigate the effect of rhein on the expression of thrombospondin-1 (TSP-1) and transforming growth factor-β1 (TGF-β1) in human renal tubular epithelial cells (HK-2). METHODS: HK-2 cells were cultured in vitro and low, medium, and high concentrations of rhein (10 μg/ml, 20 μg/ml, and 40 μg/ml) induced by AngII (1×10-7 mol/L) were observed against HK-2 TSP-1. Effects of TGF-β1 mRNA and protein expression. Real Time PCR was used to detect the expression of TSP-1 and TGF-β1 mRNA. Western blot was used to detect the expression of TSP-1 and TGF-β1. ELISA was used to detect TGF-β1 and active TGF-β1 in the culture supernatant. Results: Compared with the blank control group, AngII (1×10-7 mol/L) significantly up-regulated the mRNA expression of HK-2TSP-1 (3.84±0.48 vs 1.32±0.19; P<0.05), with TGF-β1 The mRNA expression increased (3.68±0.46 vs 1.18±0.13; P<0.05). At the same time, AngII (1×10-7mol/L) also up-regulated the expression of TSP-1 protein (0.5246±0.0936 vs 0.1015±0.0768; P<0.01), with the increase of TGF-β1 protein expression (0.5948±0.0624 vs 0.1264±0.0811; P<0.01). Compared with AngII positive control group, moderate and high concentrations of rhein significantly inhibited the expression of TSP-1, TGF-β1 mRNA (P<0.05, P<0.01) and protein expression induced by AngII (1×10-7 mol/L). (P<0.05, P<0.01). The content of total TGF-β1 and the content of active TGF-β1 in the intervention group with medium concentration of rhein decreased (P<0.05), and the content of total TGF-β1 and the content of active TGF-β1 in the intervention group with high concentration of rhein significantly decreased (P<0.05). <0.01). CONCLUSION: Rhein can inhibit the expression of TSP-1 and TGF-β1 mRNA and protein in human renal tubular epithelial cells and inhibit the secretion of TGF-β.