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、FN mRNA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

关键词:  糖肾1号方  糖尿病肾病  HK-2细胞  肾纤维化  TGF-β1

Key words:  Tangshen No.1 prescription  diabetic nephropathy  HK-2 cells  renal fibrosis  TGF-β1

摘要:

[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。

Abstract:

[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, high-sugar 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 high-sugar 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.