Effects of CTGF on Kupffer cells induced activation of hepatic stellate cells
10.3760/cma.j.issn.1007-631X.2009.02.018
- VernacularTitle:结缔组织生长因子对Kupffer细胞诱导的肝星状细胞激活的影响
- Author:
Tao LI
;
Shenban SONG
;
Xisheng LENG
;
Jiye ZHU
;
Jirun PENG
;
Yuhua WEI
- Publication Type:Journal Article
- Keywords:
Hepatic stellate cell;
Connective tissue growth factor;
RNA interference;
Kupffer cell
- From:
Chinese Journal of General Surgery
2009;24(2):142-145
- CountryChina
- Language:Chinese
-
Abstract:
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.
