复方鳖甲软肝方对LPS刺激肾小管上皮细胞MMP9／TIMP1表达的影响

Experimental Study of Effect of Compound Prescription Biejiarangan on Exprssion of MMP9/TIMPI of Tubular Epithelial Cells Stimulated by LPS

目的:研究复方鳖甲软肝方对肾小管上皮细胞LPS刺激不同时段表达MMP9、TIMP1的影响。方法:制备含药血清,将5％含药血清培养基加入体外培养的肾小管上皮细胞,分别于LPS刺激 24 h及48 h采用免疫组化及Western blot方法检测各组MMP9、TIMP1的表达。结果:LPS刺激肾小管上皮细胞12 h后,细胞胞浆内MMP9活性逐渐增强;24h达到高峰;TIMP1于LPS刺激24 h逐渐增强,48 h达到高峰上。复方鳖甲软肝方降低24h时MMP9的表达,同时也降低48h时TIMP1的表达。结论:复方鳖甲软肝方含药血清能抑制细胞早期MMP9的表达,可降低MMP9早期对基膜的破坏引起的上皮细胞转化,同时又降低后期TIMP1的表达,从而促进细胞外基质的降解,其双向调节作用可能是其防治肾间质纤维化的机制之一。

Objective To study the effect of the compound prescription Beiejiarangan of Chinese medicine on the expression of MMP9 and TIMPI of tublar epithelial cells stimulated by LPS in different period of time. Method To prepare drugcontaining serum and inject 5% culture medium with drug-contaiming serum into tubular epithelial cells cultured in vitro and then test the expression of MMP9and TIMPI of rats in all the experimental groups respectively by the methods of immunohistochemistry and Western blot in 24h and 48h respectively after the stimulation. Result The activity of MMP9 in cytoplasm gradually increases in 24h after at 24h and that of TIMPI also gradually increases stimulating tubular epithelial ceils by LPS and reaches its summit in 24h after such stimulation,but reaches its summit at 48h.This Chinese medicine is able to lower down the expression of MMP9 in 24h as well as that of TIMPI in 48h. Conclusion The drug-containing serum prepared from the compound prescription Biejiarangan is able to inhibit the expression of MMP9 of cells in their early stage and reduce epithelial-mesenchymal transformation caused by the damage of MMP9 to the basement of cells in its early stage while lowering down the expression of TIMPI in its later stage,thus advancing the degradation of cell epimatrix.The role of bilateral adjustment of this Chinese medicine as described above may be one of its mechanisms in the prevention and treatment of interrenal fibrosis.