Effect of IL-10 on the expression of HSC growth factors in hepatic fibrosis rat

瞄准：在生长因素的表示上学习 IL-10 的效果--转变生长 factor-beta1 ( TGF-beta1 )，表皮的生长因素( EGF )， hepatocyte 生长因素( HGF )和肝的纤维变性老鼠的肝的星形细胞( HSC )和外长的 IL-10 的 anti-fibrogenic 角色的导出血小板的生长因素( PDGF )。方法：肝的纤维变性被 CCl (4 ) 管理 intra-peritoneally 导致。六十只干净男 Sprague-Dawley (SD ) 老鼠随机被划分成三个组：正常控制组(GN， 8 只老鼠) ，肝的纤维变性模型组(GC， 28 只老鼠) 并且 IL-10 对待组(官方补给， 24 只老鼠) 。在第 7 和第 11 wk 的开始，在每个组的老鼠习惯性地通过一根门静脉导管与 pronase E 和类型 IV 胶原酶被酒，从肝获得的暂停被远心沉淀与 11% Nycodenz 密度坡度旋转孤立 HSC。组织学的检查被用来决定肝的纤维变性的度。RT-PCR 被采用从刚孤立的房间分析 mRNA 表示。Immunocytochemistry 被执行在主要有教养的 HSC 检测蛋白质表示。结果：老鼠肝的纤维变性随 CCl (4 ) 的注射频率的增加被开发，并且 HSC 成功地被孤立。与 GN 相比显然在在 GC 的第 7 和第 11 wk， TGF-beta1， EGF，和 HGF mRNA 增加了(P = 0.001/0.042， 0.001/0.001， 0.001/0.001 ) 并且官方补给(P = 0.001/0.007， 0.002/0.001， 0.001/0.001 ) 。为 TGF-beta1，没有差别被观察在之间官方补给并且 GN。为 EGF， mRNA 铺平在官方补给在第 7 wk 期间与 GN 相比增加了(P = 0.005 ) 并且第 11 wk (P = 0.049 ) 。为 HGF， mRNA 铺平在官方补给在第 7 wk 与 GN 相比减少了(P = 0.001 ) 并且第 11 wk (P = 0.021 ) 。两次在这些之间指，在第 7 wk 的 TGF-beta1 表示比第 11 wk 的高(P = 0.049 ) ，要不是 EGF，前者比后者低(P = 0.022 ) 。至于 PDGF mRNA，在这些组之间没有有效差量，但是差别似乎在蛋白质层次存在。由 TGF-beta1 和 EGF 的免疫细胞化学的结果是有上述调查结果的 paralleled。结论：TGF-beta1， EGF 和 HGF 的表示在肝的纤维变性老鼠的 HSC 增加了并且减少有 IL-10 的术后疗法。IL-10 由压制生长因素表示起一个 anti-fibrogenic 作用。

AIM： To study the effect of IL-10 on the expression of growth factors - transforming growth factor-β1 （TGF-β1）, epidermal growth factor （EGF）, hepatocyte growth factor （HGF）and platelet-derived growth factor （PDGF） of hepatic stellate cells （HSCs） of hepatic fibrosis rat and the anti-fibrogenic role of exogenous IL-10. METHODS： Hepatic fibrosis was induced by CCI4 administration intra-peritoneally. Sixty clean male Sprague-Dawley （SD） rats were randomly divided into three groups： normal control group （GN, 8 rats）, hepatic fibrosis model group （GC, 28 rats） and IL-10 treated group （GI, 24 rats）. At the beginning of the 7^th and 11^th wk, rats in each group were routinely perfused with pronase E and type IV collagenase through a portal vein catheter and the suspension obtained from the liver was spun by centrifugation with 11% Nycodenz density gradient to isolate HSCs. Histological examination was used to determine the degree of hepatic fibrosis. RT-PCR was employed to analyze mRNA expression from freshly isolated cells. Immunocytochemistry was performed to detect protein expression in primary cultured HSCs. RESULTS： Rat hepatic fibrosis was developed with the increase of injection frequency of CCl4, and HSCs were successfully isolated. At the 7^th and 11^th wk, TGF-β1, EGF, and HGF mRNA in GC increased obviously compared with GN （P = 0.001/0.042, 0.001/0.001, 0.001/0.001） and GI （P= 0.001/0.007, 0.002/0.001, 0.001/0.001）. For TGF-β1, no difference was observed between GI and GN. For EGF, mRNA level in GI increased compared with GN during the 7^th wk （P= 0.005） and 11^th wk （P= 0.049）. For HGF, mRNA level in GI decreased compared with GN at the 7^th wk （P = 0.001） and 11^th wk （P= 0.021）. Between these two time points, TGF-β1 expression at the 7^th wk was higher than that of the 11^th wk （P = 0.049）, but for EGF, the former was lower than the latter （P = 0.022）. As for PDGF mRNA, there was no significant difference between thesegroups, but difference seemed to exist in protein levels. Results by immunocytochemistry of TGF-β1 and EGF were paralleled with the above findings. CONCLUSION： The expression of TGF-β1, EGF and HGF increased in HSC of hepatic fibrosis rat and decreased after treatment with IL-10. IL-10 plays an anti-fibrogenic role by suppressing growth factors expression.