Shenkang Injection Solution Antagonizes the Fibrogenic Effects of Aristolochic Acid on Human Proximal Tubular Epithelial Cells in Vitro

Objective:To study whether Shenkang injection solution (SKI) can antagonize the fibrogenic effects of aristolochic acid (AA)on human proximal tubular epithelial cells (HKC) in vitro.Methods:HKC were incubated with medium alone (control group), medium containing AA-Na 10 mg/L, medium containing SKI 8 mg/ml,or medium containing AA-Na 10 mg/L and SKI 8 mg/ml, respectively. The mRNA expression of transforming growth factor-β1(TGF-β1), connective tissue growth factor(CTGF), tissue inhibitor of metalloproteinase-1(TIMP-1)and plasminogen activator inhibitor-1 (PAI-1)in cell lysate was measured by RT-PCR after 12 hours; The protein expression of TGF-β1, TIMP-1 and PAI-1 in supernate was measured by ELISA after 24 hours, and the protein expression of CTGF in cell lysate was measured by western blotting after 36 h incubation hours.Results:The mRNA and protein expression of TGF-β1,CTGF, TIMP-1 and PAI-1 was significantly up-regulated by 10 mg/L AA-Na. Compared with the control group, their the mRNA expression was up-regulated to 1.84, 1.58,1.62 and 1.29 times, respectively(P0.05), and the protein expression was up-regulated to 1.12,1.63,10.42and 1.29 times, respectively(P0.05).The up-regulated mRNA and protein expression of TGF-β1,CTGF,TIMP-1 and PAI-1 by AA-Na was significantly inhibited by 8 mg/ml SKI. Compared with the AA-Na group,the inhibition rates of mRNA expression were 41.6%,43.7%,43.8% and 24.4%, respectively(P0.05),and those of protein expression were 34.3%, 43.3%, 31.1% and 21.9%,respectively(P0.05).8 mg/ml SKI alone had no effects on both mRNA and protein expression of TGF-β1,CTGF,TIMP-1 and PAI-1.Conclusion:AA-Na can up-regulate the mRNA and protein expression of extracellular matrix(ECM)promoting synthesis factors(TGF-β1,CTGF)and inhibiting inhibit ECM degradation factors(TIMP-1, PAI-1), which can be antagonized by SKI.