The Effect Of Regulating Extracellular Signal Regulated Kinase-1 On The Biological Behaviors Of Hepatic Stellate Cells

Objective To explore the effect of regulating extracellular signal regulated kinase (ERK) -1 on the biological behaviors of hepatic stellate cells (HSC).Methods The inhibition rate of siRNA on ERK-1 expression of HSC was determined by Western blot and RT-PCR. After the ERK-1 gene silencing, the proliferation of HSC was determined with MTT assay. HSC cycle and apoptosis were measured with flow cytometery. Content of collagen typeâ…¢in the supernatants was determined by ELISA. Expression of typeâ… ,â…¢and TIMP-1 mRNA in HSC was detected by RT-PCR. The expression of leptin and collagen typeâ… andâ…¢in rat fibrotic liver by RT-PCR and immunohistochemistry respectively. Leptin-stimulating ERK, pERK expression and collagen typeâ…¢synthesis were evaluated by Western blot and ELISA respectively.Results 189 siRNA exerted a highest repression activity. After ERK-1 gene silencing, both the proliferation of HSC and expression of collagen typeâ… ,â…¢and TIMP-1 decreased compared with control group. Collagen typeâ…¢level in supernatant of HSC also decreased. The leptin expression correlated with the fibrotic degree in rodent model. Leptin could stimulate the ERK expression and typeâ…¢collagen synthesis in HSCs, which could be blocked by the ERK-1 gene silencing.Conclusions RNA interfering technique can be used as a useful tool to silence ERK-1 gene and to regulate the biological function of HSC. ERK-1 might serve as a new target for liver fibrosis therapy.