壳聚糖介导CrmA基因治疗小鼠肝纤维化的研究

目的：探讨壳聚糖介导的CrmA对小鼠肝纤维化的治疗效果，以期为肝纤维化的基因治疗提供实验基础。方法：清洁级的75只雄性小鼠随机分为正常组、模型组、壳聚糖介导的CrmA组、壳聚糖介导的空载体组、壳聚糖组，每组15只。应用30％四氯化碳橄榄油溶液3ml／kg腹腔注射制备肝纤维化小鼠模型。治疗8周后，眼眶取血，检测血清的肝功能指标，并取肝组织做HE染色，观察各组小鼠肝脏的病理形态，Real TimePCR检测肝组织IL-1β、α-SMA、TGF—β1、TIMP-1表达量。结果：与模型组小鼠相比，壳聚糖介导的CrmA组小鼠的肝纤维化程度减轻，ALT、AST显著降低（P〈0．01），肝组织IL-1β、α-SMA、TIMP1、TGF-β1的表达明显减少（P〈0．05），而模型组、壳聚糖介导的空载体组和壳聚糖组均无显著性差异。结论：壳聚糖介导的CrmA能有效减轻肝纤维化小鼠的肝脏损伤和纤维化程度，为基因治疗肝纤维化提供了一种潜在的新思路和方法。

Gene Therapy for Mouse Hepatic Fibrosis with Chitosan-mediated CrmA

Objective： To explore the therapeutic effects of the chitosan-mediated CrmA on CCl4-induced hepatic fibrosis in mouse. Methods： Seventy five male mice of clean grade were divided evenly into five groups marked ＂Normal＂, ＂Model＂, ＂chitosan-mediated CrmA＂, ＂chitosan-mediated vector＂ and ＂Chitosan＂. Liver fibrosis model mice were given 30 % carbon tetrachloride olive oil solution 3 ml/kg by intraperitoneal injection. After eight weeks of treatment, hepatic fibrosis was evaluated through measuring serum ALT and AST, examining HE pigmentation, and determining by Real Time PCR the expression levels of IL-1β, α-SMA, TGF-1β and TIMP-1 in hepatic tissue. Results： Compared with the model group, the degree of liver fibrosis in the chitosan-mediated CrmA group was obviously reduced. The ALT and AST levels of the chitosan-mediated CrmA group were much lower than that of model group （P〈0.01）. The mRNA expression of 1L-1β, α-SMA and TIMP-1 was reduced markedly in the chitosan-mediated CrmA group （P〈0.05）, as compared with the model group, whereas no significant difference was observed among the chitosan-mediated vector group, chitosan group and model group （P〉0.05）. Conclusion： The chitosan-mediated CrmA had therapeutic effects of easing the liver damage and hepatic fibrosis condition in the mouse model and provides a potential new approach for the treatment of liver fibrosis.