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目的探讨中药方剂益肾活血汤通过p38丝裂原活化蛋白激酶(p38MAPK)信号转导途径对肾小球系膜细胞纤连蛋白(FN)、Ⅳ型胶原(COLⅣ)表达的影响。方法采用血清药理学方法制备中药血清,体外培养肾小球系膜细胞分组为:对照组(正常鼠血清)I、L-1β组(正常鼠血清+10%IL-1β)、中药血清组(中药血清+10%IL-1β)。采用双抗体夹心ELISA法检测大鼠系膜细胞FN、COLⅣ表达,蛋白印记法(Western blot)检测磷酸化p38MAPK(p-p38MAPK)及α-平滑肌肌动蛋白(-αSMA)水平。结果与IL-1β组相比,中药血清组FN、p-p38MAPK蛋白的表达量降低(P<0.01)。-αSMA、COLⅣ的表达亦降低(P<0.05)。结论中药益肾活血汤可降低肾小球系膜细胞-αSMA的表达,从而抑制肾小球系膜细胞的表型转化,减少细胞外基质FN、COLⅣ的表达,该作用可能是通过抑制p-p38MAPK信号通路而实现的。
Objective To investigate the effect of Yishen Huoxue Decoction, a traditional Chinese medicine prescription, on the expression of fibronectin (FN) and type IV collagen (COLIV) in glomerular mesangial cells through the signal transduction pathway of p38 mitogen-activated protein kinase (p38MAPK). Methods The serum of traditional Chinese medicine was prepared by serum pharmacology. In vitro culture of glomerular mesangial cells was divided into control group (normal mouse serum) I, L-1β group (normal mouse serum +10% IL-1β) and Chinese medicine serum group ( Chinese medicine serum +10% IL-1β). The expression of FN and COLIV in rat mesangial cells was detected by double antibody sandwich ELISA. The levels of phosphorylated p38MAPK (p-p38MAPK) and α-smooth muscle actin (-αSMA) were detected by Western blot. Results Compared with the IL-1β group, the expression of FN and p-p38MAPK protein in the serum of the Chinese medicine group was decreased (P<0.01). -αSMA and COLIV expression also decreased (P<0.05). Conclusion Yishen Huoxue Decoction can reduce the expression of α-SMA in glomerular mesangial cells, inhibit the phenotypic transformation of glomerular mesangial cells and reduce the expression of extracellular matrix FN and COLIV. This effect may be inhibited by p- p38MAPK signaling pathway is realized.