Hepatocyte growth-promoting factor partially reverses monocyte chemotatic protein-1- and aristolochic acid I - Induced epithelial-to-mesenchymal transition of human kidney epithelial cells / 第二军医大学学报

ABSTRACT

Objective: To observe the influence of hepatocyte growth-promoting factor (pHGF) on monocyte chemotatic protein-1-(MCP-1) and aristolochic acid I (AA I )-induced epithelial-to-mesenchymal transition (EMT) and apoptosis of human kidney epithelial cell line(HKC). Methods: The HKC cells were randomly divided into blank control group (control groups), epithelial-to-mesenchymal transition model group (model group), and pHGF inhibition group (pHGF groups) with pHGF at different concentrations (0. 15, 1. 5, 15, 150, and 1 500 ng/ml). The EMT model was established by exposing HKC cells to MCP-1 (0. 1 μg/ml)and AA I (10 μg/ml). Cells in the pHGF groups were the model cells treated with different concentrations of pHGF. Cells in the control group were cultured routinely. WST-8 method and flow cytometry were used to observe the proliferation and apoptosis of HKC cells, respectively. The mRNA expression of α-smooth muscle actin(α-SMA) was determined by reverse transcriptase-polymerase chain reaction (RT-PCR), and the expression of α-SMA, fibronectin (FN), and transforming growth factor-β1 (TGF-β1) in HKC cells were assessed by indirect enzyme immunohistochemistry. Results: The cell inhibitory rate, apoptotic rate, and expression of α-SMA mRNA were significantly increased in the model group and pHGF groups compared with those in the control group (P<0. 01), indicating the successful establishment of EMT model. Compared with the model group, pHGF at 150 ng/ml, but not at other concentrations, significantly decreased the inhibition rate of HKC cells(P<0. 01). The apoptotic rate of HKC cells in all the pHGF groups were significantly lower than that in the model group (P<0. 01), pHGF at 150 ng/ml also greatly decreased the expression of α-SMA mRNA, and significantly down-regulated the expression of α-SMA, TGF-β1, and FN protein. Conclusion: pHGF at 150 ng/ml can partly reverse MCP-1- and AA I -induced HKC cell growth inhibition, apoptosis, and EMT.