糖耐康对转化生长因子-β_1诱导的HK-2细胞分泌细胞外基质成分的影响

Effect of Tangnaikang on Extracellular Matrix Components of Human Renal Tubular Epithelial Cell HK-2 Induced by TGF-β_1

目的观察糖耐康对转化生长因子-β1(TGF-β1)诱导的人肾小管上皮细胞(HK-2)对细胞外基质成分表达的影响,探讨糖耐康治疗肾间质纤维化的作用机制。方法将HK-2细胞用含10%胎牛血清的DMEM/F12(1∶1)培养基培养,并分为6组:空白对照组、TGF-β1诱导组(TGF-β110 ng/mL)、空白血清对照组(TGF-β110 ng/mL+10%空白血清)、干预1组(TGF-β110 ng/mL+5%糖耐康药物血清)、干预2组(TGF-β110 ng/mL+10%糖耐康药物血清)、干预3组(TGF-β110 ng/mL+20%糖耐康药物血清)。药物干预24 h,荧光定量PCR检测Ⅰ型胶原(ColⅠ)、Ⅲ型胶原(ColⅢ)和纤连蛋白(FN)的mRNA表达。结果 HK-2细胞经TGF-β1诱导后,ColⅠ、ColⅢ和FN的mRNA表达显著上升,与空白对照组比较,差异有统计学意义(P<0.05)。经糖耐康药物血清干预后,其表达逐步下降,与TGF-β1诱导组比较,差异有统计学意义(P<0.05)。而空白血清无此作用。结论糖耐康在一定程度上能够抑制TGF-β1诱导的人肾小管上皮细胞ColⅠ、ColⅢ和FN的mRNA表达,减少细胞外基质成分的分泌,具有防治肾间质纤维化的作用。

Objective To explore the effect of Tangnaikang（TNK） on extracellular matrix expression of human tubular epithelial cell HK-2 induced by TGF-β1 and explore its mechanism on the renal fibrosis.Methods The HK-2 cells were cultured by DMEM/F12（1：1） with 10% fetal bovine serum and divided into control group,TGF-β1 group（TGF-β110 ng/mL）,rat serum control group（TGF-β110 ng/mL ＋10% rat serum）,TNK-containing rat serum therapy groups（TGF-β110 ng/mL＋5% Tangnaikang,or ＋10% Tangnaikang,or ＋20% TNK）.After 24 h of administration,the expression of ColⅠ,Col Ⅲ and FN mRNA were tested by fluorescence quantitative PCR assay.Results The expression of ColⅠ,Col Ⅲ and FN mRNA of HK-2 cultured with TGF-β1 were much higher than the control,and significantly decreased in HK-2 cultured with TGF-β1 plus Tangnaikang compared with only TGF-β1（P〈0.05）,but rat serum control had no effect.Conclusion TNK could inhibit the expression of ColⅠ,Col Ⅲ and FN mRNA of HK-2 cell induced by TGF-β1,and prevent the development of renal fibrosis to some extent.