Effects of CTGF on Kupffer cells induced activation of hepatic stellate cells

Objective To investigate the effect of RNA interference connective tissue growth factor (CTGF)expression on Kulclffer cells(KC)induced activation of hepatic stellate cells(HSC).Methods Rat CTGF RNA interference vector Psilencer 3.1H1-Neo-CTGF was constructed and identified.HSC cell line rHSC-99 cells were divided into three groups,group A served as control,group B transfected with vector without CTGF interference.group C was RNA interferenee CTGF expression of HSC.RT-PCR was used to measure the expression of CTGF in HSC.Rat Kur)ffer cells were isolated and identified.and cocultured with HSC in the 3 groups respectively.MTT assay was used to evaluate the proliferation of HSC.RT-PCR was used to measure the expression of TGF-β1 and precollagen type I in HSC.Western blot was used to measure the expression of TGF-β1 in HSC.ELISA was used to detect the production of precollagen type I protein.Immunofluorescence was used to detect the expression of ot-smooth muscle actin(α-SMA)in HSC.Resuits After CTGF RNA interference vector transfection.CTGF expression of HSC decreased by 22%(P＜0.01).The yield rate of Kupffer cell was 5×107 and the cell viability exceeded 98%.In the HSC and KC co-culture system.the proliferation and activation of HSC were inhibited while RNA interferenee CTGF of HSC.As compared with control,HSC proliferation decreased by 29%(P＜0.01).Precollagen type I and ot-SMA expression decreased by 38%(P＜0.01).Production of precollagen type I protein in culture medium decreased bv 48%(P＜0.01).Conclusions Blockade CTGF expression of HSC inhibits KC induced activation of HSC. Key words: Hepatic stellate cell; Connective tissue growth factor; RNA interference; Kupffer cell