---

作 者 ：刘明龙,曾永祥,卢守燕,李建华,刘天喜

期 刊 ：中国中药杂志 2011年 36卷 18期 页码：2571.

关键词：复肾颗粒；人肾小管上皮细胞；细胞增殖；肾纤维化；转化生长因子β激活激酶；

Keywords:fushenkeli, human renal tubular epithelial cells, cell proliferation, renal fibrosis, TGFβ-activated kinase 1,

摘 要 ：目的: 探讨复肾颗粒对转化生长因子-β1(TGF-β1)诱导下人肾小管上皮细胞(HK-2)增殖时转化生长因子β激活激酶(TAK1)表达的影响及其可能机制。 方法 :体外培养HK-2细胞,分为对照组,TGF-β1诱导组(5 μg·L^-1)、干预1组(TGF-β1 5 μg·L^-1+复肾颗粒100 μg·L^-1)、干预2组(TGF-β1 5 μg·L^-1+复肾颗粒500 μg·L^-1)、干预3组(TGF-β1 5 μg·L^-1+复肾颗粒1 g·L^-1)。培养12,24,48 h 以MTT法检测细胞增殖情况,ELISA法检测细胞上清IV型胶原蛋白(Col IV)含量,实时荧光定量PCR检测TAK1 mRNA的相对含量,Western印迹法检测TAK1蛋白表达。 结果 :①TGF-β1组细胞的增殖及Col IV的表达显著高于对照组(P<0.05,P<0.01),复肾颗粒干预后细胞增殖和Col IV的表达显著低于TGF-β1组(P<0.05,P<0.01)。②TGF-β1诱导12 h细胞TAK1表达开始上调,且各时间点表达量均高于对照组(P<0.01),复肾颗粒干预后,与同时间点TGF-β1组比较显著下调(P<0.01),尤以干预3组为甚。 结论 :在体外,复肾颗粒对TGF-β1诱导HK-2细胞增殖过程中TAK1蛋白及mRNA的表达有下调作用,其可能是通过激活肾小管上皮细胞某些抑制因子抑制TAK1转录和表达。

Abstract:Objective : To investigate the effects of fushenkeli on the expression of TAK1 in the proliferation of the renal tubular epithelial cells induced by TGF-β1 and its possible mechanism. Method : Human renal tubular epithelial (HK-2) cells were divided into five groups:blank control group, TGF-β1 group (5 μg·L^-1),intervention group 1 (5 μg·L^-1 of TGF-β1+100 mg·L^-1 of fushenkeli), intervention group 2 (5 μg·L^-1 of TGF-β1+500 mg·L^-1 of fushenkeli) and intervention group 3 (5 μg·L^-1 of TGF-β1+1 g·L^-1 of fushenkeli).HK-2 proliferation was detected by methyl thiazolyl tetrazolium (MTT) assay.Type Ⅳ collagen in the supernatants of the cultured HK-2 was detected by ELISA at 12,24,48 hours respectively.The protein and mRNA expressions of TAK1 was measured by Western blot and real-time quantitative PCR. Result : ①The cell proliferation and the expression of type Ⅳ collagen were increased compared with the control group (P<0.05,P<0.01),but they were decreased in intervention group. ②The expressions of protein and mRNA of TAK1 in TGF-β1 group were upregulating significantly compared with control group (P<0.01),but they were downregulating in intervention group,especially in intervention group 3. Conclusion : Fushenkeli could inhibits TAK1 expression induced by TGF-β1 in the proliferation of HK-2 cell.

---