Shi Mei-Na, Zheng Wei-Da, Zhang Li-Juan, Chen Zhi-Xin, Wang Xiao-Zhong
Department of Gastroenterology, Union Hospital of Fujian Medical University, Fuzhou 350001, Fujian Province, China.
World J Gastroenterol. 2005 Aug 21;11(31):4788-93. doi: 10.3748/wjg.v11.i31.4788.
To study the effect of IL-10 on the expression of growth factors--transforming growth factor-beta1 (TGF-beta1), 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.
Hepatic fibrosis was induced by CCl(4) 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 7th and 11th 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.
Rat hepatic fibrosis was developed with the increase of injection frequency of CCl(4), and HSCs were successfully isolated. At the 7th and 11th wk, TGF-beta1, 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-beta1, no difference was observed between GI and GN. For EGF, mRNA level in GI increased compared with GN during the 7th wk (P = 0.005) and 11th wk (P = 0.049). For HGF, mRNA level in GI decreased compared with GN at the 7th wk (P = 0.001) and 11th wk (P = 0.021). Between these two time points, TGF-beta1 expression at the 7th wk was higher than that of the 11th 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 these groups, but difference seemed to exist in protein levels. Results by immunocytochemistry of TGF-beta1 and EGF were paralleled with the above findings.
The expression of TGF-beta1, 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.
研究白细胞介素-10（IL-10）对肝纤维化大鼠肝星状细胞（HSCs）中生长因子——转化生长因子-β1（TGF-β1）、表皮生长因子（EGF）、肝细胞生长因子（HGF）和血小板衍生生长因子（PDGF）表达的影响以及外源性IL-10的抗纤维化作用。
通过腹腔注射四氯化碳（CCl₄）诱导肝纤维化。60只清洁级雄性Sprague-Dawley（SD）大鼠随机分为三组：正常对照组（GN，8只大鼠）、肝纤维化模型组（GC，28只大鼠）和IL-10治疗组（GI，24只大鼠）。在第7周和第11周开始时，通过门静脉导管对每组大鼠常规灌注链霉蛋白酶E和IV型胶原酶，将肝脏获得的悬液用11% Nycodenz密度梯度离心分离HSCs。采用组织学检查确定肝纤维化程度。运用逆转录-聚合酶链反应（RT-PCR）分析新鲜分离细胞的mRNA表达。进行免疫细胞化学检测原代培养HSCs中的蛋白表达。
随着CCl₄注射频率增加，大鼠肝纤维化形成，且成功分离出HSCs。在第7周和第11周，与GN组（P = 0.001/0.042、0.001/0.001、0.001/0.001）和GI组（P = 0.001/0.007、0.002/0.001、0.001/0.001）相比，GC组中TGF-β1、EGF和HGF mRNA明显增加。对于TGF-β1，GI组和GN组之间未观察到差异。对于EGF，在第7周（P = 0.005）和第11周（P = 0.049）时GI组mRNA水平高于GN组。对于HGF，在第7周（P = 0.001）和第11周（P = 0.021）时GI组mRNA水平低于GN组。在这两个时间点之间，第7周时TGF-β1表达高于第11周（P = 0.049），但对于EGF，前者低于后者（P = 0.022）。至于PDGF mRNA，这些组之间无显著差异，但蛋白水平似乎存在差异。TGF-β1和EGF的免疫细胞化学结果与上述发现一致。
肝纤维化大鼠HSCs中TGF-β1、EGF和HGF表达增加，IL-10治疗后降低。IL-10通过抑制生长因子表达发挥抗纤维化作用。
