糖肾1号方含药血清对高糖环境下培养的HK-2细胞中TGF-β1、α-SMA、FNmRNA和蛋白水平的影响

Effect of Tangshen No.1 prescription contained serum on the levels of TGF-β1, α-SMA, FN mRNA and protein in HK-2 Cells cultured in high glucose environment

[目的]观察糖肾1号方含药血清干预高糖环境下培养的人肾小管上皮细胞(HK-2)中转化生长因子-β1(TGF-β1)、α-平滑肌蛋白(α-SMA)、纤维连接蛋白(FN)mRNA和蛋白水平,初步探索糖肾1号方治疗糖尿病肾病可能的作用机制。[方法]选用HK-2细胞,分为空白组、高糖组、空白血清组、糖肾1号方含药血清低剂量组、糖肾1号方含药血清高剂量组。高糖环境下培养24 h后,进行血清干预24 h,培养48 h后收集细胞,通过逆转录-聚合酶链反应(RT-PCR)和蛋白免疫印迹(Western Blot)法检测HK-2细胞中TGF-β1、α-SMA、FN mRNA和蛋白表达水平。[结果]1)与空白组相比,高糖组的TGF-β1、α-SMA、FN mRNA和蛋白表达水平明显增多,差异具有统计学意义(P<0.05)。2)与高糖组相比,糖肾1号方低剂量组、糖肾1号方高剂量组TGF-β1、α-SMA、FN mRNA表达水平减少,差异具有统计学意义(P<0.05)。3)与高糖组相比,糖肾1号方低剂量组FN蛋白表达水平减少,差异具有统计学意义(P<0.05),糖肾1号方高剂量组α-SMA、FN蛋白表达水平减少,差异具有统计学意义(P<0.05)。4)与糖肾1号方低剂量组相比,糖肾1号方高剂量组TGF-β1、FN mRNA降低,差异具有统计学意义(P<0.05)。[结论]糖肾1号方含药血清可抑制TGF-β1、α-SMA、FN mRNA和α-SMA、FN蛋白表达水平,糖肾1号方高剂量组效果优于糖肾1号方低剂量组。

[Objective] To observe the effect of Tangshen No.1 prescription on the mRNA and protein levels of TGF-β1, α-SMA, and FN in HK-2 cells cultured under high glucose conditions, and preliminary explore the possible mechanism of Tangshen No.1 prescription in the treatment of diabetic nephropathy. [Methods] Renal tubular epithelial cells（HK-2） were selected and divided into blank group, highsugar group, blank serum group, low-dose group of Tangshen No.1 prescription-containing serum, and high-dose group of Tangshen No.1 prescription-containing serum. Cells were performed in a high glucose environment for 24 hours; serum was intervened for 24 hours. Cells were harvested after 48 hours of co-culture. Use RT-PCR and western blotting methods detected TGF-β1, α-SMA, FN mRNA and protein expression levels in HK-2 cells. [Results] 1） Compared with the blank group, the expression levels of TGF-β1, α-SMA, FN mRNA and protein in the high glucose group increased significantly, the difference was statistically significant（P〈0.05）. 2） Compared with the highsugar group, the expression levels of TGF-β1, α-SMA and FN mRNA in the low-dose group of Tangshen No.1 and high-dose group of Tangshen No.1 were significantly decreased, and the difference was statistically significant（P〈0.05）. 3） Compared with the high-sugar group, the expression of FN protein in the low-dose group of Tangshen No.1 low-dose group was significantly reduced（P〈0.05）. α-SMA and FN protein in high-dose group of Tangshen No.1. The expression level was significantly reduced and the difference was statistically significant（P〈0.05）. 4） Compared with the low serum concentration group, the serum TGF-β1 and FN levels in high-concentration sera group decreased more significantly（P〈0.05）. [Conclusion] The Tangshen No.1 prescription serum significantly inhibited the expression of TGF-β1, α-SMA, FN mRNA and α-SMA and FN protein, and the effect of high concentration of drug-containing serum was superior to that of low-concentration drug-containing serum.