基于HK-2细胞转分化研究益肾化瘀方抗肾间质纤维化机制

Study on the mechanism of Yishen Huayu Formula against renal interstitial fibrosis based on the transdifferentiation of HK-2 cells

目的:观察益肾化瘀方含药血清对人近端肾小管上皮细胞株(HK-2)细胞转分化过程中γ干扰素(IFN-γ)、转化生长因子-β1(TGF-β1)、Ⅰ型胶原(Col-Ⅰ)、Ⅳ型胶原(Col-Ⅳ)表达的影响,探究该方抗肾间质纤维化机制。方法:将体外培养的HK-2细胞随机分为:正常对照组,缬沙坦含药血清组,AngⅡ刺激组,5%、10%、20%益肾化瘀方含药血清组,分别干预12、24、48 h。检测各组细胞IFN-γ、TGF-β1、Col-Ⅰ、Col-Ⅳ的表达情况。结果:与正常对照组同期比较,AngⅡ刺激组IFN-γ表达量显著降低(P<0.05),TGF-β1、Col-Ⅰ、Col-Ⅳ表达量显著升高(P<0.05);与AngⅡ刺激组同期比较,10%、20%益肾化瘀方含药血清组及缬沙坦组IFN-γ表达量显著升高(P<0.05),各治疗组IFN-γ表达量显著降低(P<0.05);其中20%益肾化瘀方含药血清组与缬沙坦含药血清组各指标表达量无显著性差异。结论:益肾化瘀方含药血清可通过上调IFN-γ及下调TGF-β1、Col-Ⅰ、Col-Ⅳ的表达保护肾脏。

Objective: To observe the effects of Yishen Huayu Formula serum containing on the expression of interferon γ(IFN-γ), transforming growth factor β1(TGF-β1), type Ⅰ collagen(Col-Ⅰ) and type Ⅳ collagen(Col-Ⅳ) in HK-2 cells during transdifferentiation, and explore the mechanism of Yishen Huayu Formula against interstitial fibrosis. Methods: HK-2 cells cultured in vitro were randomly divided into: normal control group, Valsartan medicated serum group, AngⅡ stimulation group and 5%, 10%, 20% Yishen Huayu Formula medicated serum group, which were intervened for 12 h, 24 h and 48 h respectively.The expression of IFN-γ, TGF-β1, Col-Ⅰ and Col-Ⅳ in each group were detected. Results: Compared with the normal control group, the expression of IFN-γ in AngⅡ stimulation group decreased significantly(P<0.05), while the expression of TGF-β1,Col-Ⅰ and Col-Ⅳ increased significantly(P<0.05);Compared with AngⅡ stimulation group, the expression of IFN-γ in 10%and 20% Yishen Huayu Formula medicated serum group and Valsartan medicated serum group increased significantly(P<0.05),while the expression of IFN-γ decreased significantly(P<0.05);There was no significant difference in the expression of each index between 20% Yishen Huayu Formula medicated serum group and Valsartan medicated serum group. Conclusion: The serum containing Yishen Huayu Formula can protect the kidney by up-regulating IFN-γ and down-regulating the expression of TGF-β1, Col-Ⅰ and Col-Ⅳ.