Cenicriviroc治疗肝纤维化的实验研究

Therapeutical effect of cenicriviroc on liver fibrosis

探讨Cenicriviroc(CVC)对硫代乙酰胺(TAA)所致大鼠肝纤维化的治疗作用。SD雄性大鼠66只被随机分为模型对照组和CVC治疗组。各组大鼠于同日开始腹腔注射TAA 30mg/Kg,3次/周,连续8周;另设空白对照组。治疗组分别在TAA诱导肝纤维化不同时间,按30mg·kg-1·d-1灌胃给药,包括:与TAA同日开始给CVC,连续8周(1B组);TAA腹腔注射4周后,给予CVC,连续4周(2B组)和TAA腹腔注射8周停用后,再给予CVC,4周(3B组)。治疗结束后,检测血清中生化指标,留取肝组织,行HE和苦味酸天狼星红溶液染色,观察肝脏病理学改变,并进行肝纤维化定量分析。同时,以qRT-PCR检测CVC对培养肝星状细胞(HSC)表达α-平滑肌肌动蛋白(α-SMA)和胶原蛋白I(collagen I)的直接作用。与模型组比,早、中期CVC治疗(1B组和2B组)显著降低肝纤维化大鼠增高的肝脏系数(P<0.05)和血清中异常升高的天冬氨酸转氨酶(AST)、丙氨酸转氨酶(ALT)(P<0.01),病理组织学检查结果表明,1B组和2B组大鼠肝纤维组织增生和肝组织炎症活动度较模型组均有显著的改善(P<0.05),其肝组织中胶原纤维面积显著降低(P<0.01),但是CVC对已形成的肝硬化(3B组)无显著改善作用。体外实验显示,CVC可以直接抑制HSC胶原蛋白I mRNA表达(P<0.05),但α-SMA mRNA表达与对照组无明显差异。CVC具有明显抑制肝纤维化形成作用,但对已形成的肝硬化缺乏逆转作用。

The prurpose of this study was to investigate therapeutic effect of cenicriviroc （CVC） on TAA-induced liver fibrosis in rats. Sixty-nine male SD rats were randomly divided into control group, model group and CVC-treated group. The rats of the model group and the treatment group began to receive intraperitoneal injections of TAA at close of 30mg/Kg, 3 times per week for consecutive 8 weeks. At the same time the model group was given vehicle for 8 weeks. Rats of the treatment group were given CVC at different stages of liver fibrosis. CVC was administered daily at doses of 30 mg/kg/day by gavage either at the beginning of TAA administration for 8 weeks（1B group）, during weeks 4-8（2B group）, or from weeks 8-12（3B group）. Then serum levels of biochemical indicators and liver specimens were collected. Hepatic pathology was performed by hematoxylin-eosin staining and Siru red staining to detect the areas of collagen fiber in the liver. Treatment with 10nmol/L CVC for 24 h and 48 h markedly inhibited expression of collagen Ⅰ, mRNA, but not of SMA. In vivo, as compared with in model groups, CVC significantly decreased serum levels of aspartate aminotransferase （AST）, alanine aminotransferase （ALT） and inhibited liver fibrosis when started concurrently with TAA and during weeks 4-8 with TAA. In contrast, there was no significant effect of CVC on liver fibrosis when begun after cirrhosis was already established. Therefore, CVC has antifibrotic effect towards human HSC in culture, and is a potent antifibrotic agent in TAA-indueed hepatic fibrosis in rats. The drug is effective at the early stage of liver fibrosis, but is not effective when cirrhosis has already been established.