Effect of lumbrokinase on the synthesis of Src and FAK induced by TGF-β1 in human renal tubular epithelial cells

Abstract

Objective To observe the regulation of lumbrukinase on the expression of α-smooth muscle actin, fibronectin, focal adhesion kinase and Src kinase induced by transforming growth factor β1 in human renal tubular epithelial cells. Methods According to random number table method, the human renal tubular epithelial cells were divided into the normal group, TGF-β1 model group, benazepril group, lumbrukinase low, medium and high dose group. Except the normal group, the other groups were treated with TGF-β1 10 μg/ml. After 30 minutes, the benazepril group added benazepril 10 μmol/L, and the low, medium, high dose groups of lumbrukinase were respectively treated with lumbrokinase 30, 60, 120 U/ml to intervene. After 24 hours of cultivation. Western blotting and real-time PCR were used to detect the expression of α-SMA, FN, FAK and Src. Results Compared with the model group, the expressions of α-SMA (0.84 ± 0.14, 0.72 ± 0.08, 0.69 ± 0.05 vs. 1.24 ± 0.03) and FN (0.59 ± 0.09, 0.55 ± 0.11, 0.44 ± 0.08 vs. 0.83 ± 0.18) and FAK (0.94 ± 0.04, 0.79 ± 0.05, 0.70 ± 0.02 vs. 1.29 ± 0.07) and Src (0.87 ± 0.20, 0.78 ± 0.15, 0.71 ± 0.11 vs. 1.23 ± 0.01) proteins in the high doses of lumbrical kinase group were significantly lower than those in the model group (P<0.05), the expressions of α-SMA (3.13 ± 0.62, 2.76 ± 0.14, 2.15 ± 0.33 vs. 4.12 ± 0.32) and FN (3.08 ± 0.34, 2.78 ± 0.17, 2.49 ± 0.11 vs. 4.34 ± 0.06) and FAK (1.73 ± 0.23, 1.63 ± 0.36, 1.57 ± 0.27 vs. 2.61 ± 0.59) and Src (2.11 ± 0.17, 1.78 ± 0.25, 1.71 ± 0.22 vs. 2.78 ± 0.47) mRNA in the high doses of lumbrical kinase group decreased significantly (P<0.05). Conclusions Lumbrokinase may prevent the development of renal fibrosis by regulating the expression of FN, FAK and and reducing the production of α-SMA, FN. Key words: Fibrosis; Lumbrokinase; α-smooth muscle actin; src-Family kinases; Epithelial- mesenchymal transition; Epithelial cells