疏肝健脾方对四氯化碳诱导小鼠肝纤维化模型影响

Effect of Shugan Jianpi Recipe on Mice Models of Liver Fibrosis Induced by Carbon Tetrachloride

目的:观察疏肝健脾方对四氯化碳(Carbon tetrachloride,CCl4)诱导的小鼠肝纤维化模型的防治作用。方法:小鼠随机分为正常组,模型组,疏肝健脾高、低两个剂量组。除正常组外,其余各组均腹腔注射10%CCl4溶液2 m L/kg体重造模。自造模之日起,两个实验组分别给予疏肝健脾方提取液按1 g生药/m L和3 g生药/m L的剂量灌胃共8周,正常组及模型组给予等量生理盐水。于末次戊巴比妥麻醉小鼠,腹主动脉取血,离心取上清,检测血清丙氨酸转氨酶(ALT)、天门冬氨酸转氨酶(AST)含量以及透明质酸(HA)含量,摘取肝脏、脾脏,检测肝脏器系数、脾脏器系数、肝组织匀浆中羟脯氨酸(Hyp)含量、Western印迹检测肝组织匀浆中TIMP-2及MMP-2的表达,观察疏肝健脾方对上述指标的影响,电镜、光镜下观察大鼠肝脏病理学检查结果。结果:疏肝健脾方预防用药能不同程度降低小鼠肝脏器比、降低血清转氨酶ALT水平、HA含量,降低肝组织Hyp含量,增加MMP-2表达及抑制TIMP-2表达的作用,光镜及电镜下显示疏肝健脾方具有较好的保护肝细胞,减轻四氯化碳对大鼠肝组织破坏、防止胶原纤维增生、沉积的作用,其中以高剂量组按3 g生药/m L给药效果最为明显。结论:疏肝健脾方对CCl4诱导小鼠肝纤维化模型具有防治作用。其可以通过降低小鼠肝脏器比,降低血清转氨酶ALT、AST水平、HA含量,降低肝组织Hyp含量,增加肝组织匀浆MMP-2表达及抑制肝组织匀浆TIMP-2表达,从而达到治疗肝纤维化的目的。

Objective： To observe the prevention and effect of Shugan Jianpi Recipe on mice models of liver fibrosis induced by carbon tetrachloride （ CCI4 ）. Methods ： The mice were randomly divided into the normal group, the model group, the high dose group and the low dose group. The rest three groups were induced by intraperitoneal injection of 10% carbon tetrachloride 2 mL/kg to duplicate the liver fibrosis model except the normal group. Since the day of the animal model, all the groups were filled with the Shugan Jianpi Recipe extracts by 1 g/mL and 3 g/mL. The normal group and the model group were given equat volume of saline. Finally all the mice were anesthesia with pentobarbital mice, taking blood from the abdominal aorta, getting centrifugal supernatant, detecting serum alanine aminotransferase （ALT）, aspartate aminotransferase （ AST ） and hyaluronic acid （ HA ） content, removing the liver, spleen, counting the liver detection coefficient and the coefficient spleen, liver hydroxyproline （ Hyp ）, matrix metalloproteinases-2 （ MMP-2 ） and tissue inhibitor of metalloproteinase-2 （ 2 ） content to observe the effect of Shugan Jianpi Recipe on the index above-mentioned. Histological changes of hepatic tissue were under light microscope. Results ： Compared with the model group, high dose group can reduce the liver detection coefficient, serum ALT activity and HA concentration, lowing down the levels of Hyp of liver tissue by 3 g crude drug/mL. Conclusion ： Shugan Jianpi Recipe can prevent and treat rats model of CCl4-induced liver fibrosis through descending the liver detection coefficient, serum ALT, serum AST, HA content, Hyp in the liver tissue and TIMP-2 level, ascending MMP-2 level .