大黄素对高糖介导的大鼠NRK52E TSP1和TGF-β_1表达的影响

Effect of rhein on expression of thrombospondin-1and transforming growth factorβ_1 in NRK52E cells stimulated by high glucose

目的:探讨大黄素对高糖介导的大鼠肾小管上皮细胞(NRK52E)血小板反应蛋白1(thrombospondin-1,TSP1)和转化生长因子β1(transforming growth factorβ1,TGF-β1)表达的影响。方法:在NRK52E体外培养的基础上,观察高糖(30mmol/L)诱导下低、中和高浓度的大黄素(10ug/ml、20ug/ml和40ug/ml)对NRK52E TSP1、TGF-β1的mRNA和蛋白表达的影响。采用Real Time PCR检测TSP1、TGF-β1mRNA表达,采用Western blot检测TSP1、TGF-β1蛋白表达。总TGF-β1和活性TGF-β1检测采用ELISA法。结果:与空白对照组比较,高糖(30mmol/L)明显上调了NRK52E TSP1的mRNA表达(3.34±0.18 vs 1.12±0.13;P<20.05)、伴有TGF-β1的mRNA表达增加(3.52±0.22 vs 1.24±0.09;P<20.05)。同时高糖(30mmol/L)也上调了TSP1蛋白表达(0.5184±0.0736 vs 0.1204±0.0376;P<20.01),伴有TGF-β1蛋白表达的增加(0.5698±0.0324 vs 0.1032±0.0322;P<20.01)。与高糖阳性对照组比较,中浓度和高浓度大黄素明显抑制了高糖(30mmol/L)诱导的TSP1、TGF-β1的mRNA(P<20.05;P<20.01)和蛋白表达(P<0.05;P<20.01)。中浓度大黄素干预组总的TGF-β1含量和活性TGF-β1含量均减少,(P<20.05);高浓度大黄素干预组总的TGF-β1含量和活性TGF-β1含量均明显减少(P<20.01)。结论:大黄素能抑制大鼠肾小管上皮细胞TSP1、TGF-β1的mRNA和蛋白表达,并下调总的和活性TGF-β1。

Objective：To investigate the effect of rhein on expression of thrembospondin -1 and transforming growth factor β1 in NRK52E cells stimulated by high glucose.. Methods：NRK52E cells were stimulated by different concentrations of rhein（ 10,20,40ug/ ml } in the presence of high glueose（30mmol/L） , untreated NRK52E ceils were acted as normal control group, NRK52E cells were stimulated by high glucose（30mmol/L） were acted as positive group. Real Time PCR was used to detect the expression of TSPlmRNA and TGF - β1 mRNA . Western blot analysis was used to detect the expression of TSP1 and TGF - β1 protein. ELISA was used to detect the levels of total and active TGF - β1. Results：The expression of TSP1 mRNA 13.34 ± 0.18 vs 1.12 ± 0.13 ; P 〈 20.05 ） and TGF - β1 mRNA （ 3.52 ± 0.22 vs 1.24 ± 0.09 ; P 〈 20.05 ） were up - regulated obviously when stimulated by high glucose （ 30mmol/L ）. The protein expression of TSP1 （ 0.5184 ± 0. 0736 vs 0. 1204 ± 0. 0376 ; P 〈 20.01 ） and TGF - β1 （0- 5698 ± 0. 0324 vs 0. 1032 ± 0. 0322 ; P 〈20.01 ） were also up - regulated simultaneously when stimulated by high glucose（ 30mmol/L）. The expression of TSP1 and TGF - β1 incueed by high glucase（30mmol/L） were clown - regulated markedly with 20ug/ml and 40ug/ml of rhein in mRNA and protein levels（ P 〈 20.05, P 〈 20.01 ）. And rhein （ 20ug/ml and 40ug/ml ） could reduce the levels of total and active TGF - β1 （ P 〈 20.05, P 〈 20.01 ）. Conclusion： Rhein can inhibit the expression of TSP1 and TGF - β1 in mRNA and protein levels and clown -regulate the levels of total and active TGF - β1.