胶原酶Ⅱ对肝硬化大鼠的治疗作用

Therapeutic effect of collagenase Ⅱ against rat liver cirrhosis

目的:探讨应用腺病毒与乙型肝炎病毒嵌合载体表达胶原酶Ⅱ对肝硬化大鼠模型的治疗作用.方法:以硫代乙酰胺口服法16 wk诱导大鼠形成肝硬化模型,用腺病毒穿梭质粒与乙型肝炎病毒嵌合载体表达截断的胶原酶Ⅱ基因tMMP-8片断或全长MMP-8基因,分别构建了腺病毒Ad-CH-tMMP8、Ad-C-MMP8,并以表达红色荧光蛋白(red fluorescent protein,RFP2)的Ad-CH-RFP2作为对照,经尾静脉注射治疗已形成肝硬化模型的大鼠.结果:在Ad-CH-tMMP8、Ad-C-MMP8尾静脉注射肝硬化大鼠的体内转染研究中,转染4 wk相比于模型组,肝纤维化程度显著的减轻,肝细胞再生明显,同时伴有肝组织羟脯氨酸含量的下降(28.97μg/g±2.36μg/g vs 17.04μg/g±0.61μg/g,17.62μg/g±1.30μg/g,P<0.05),AdCH-RFP2对照组,纤维化程度同模型组类似.结论:用腺病毒与乙型肝炎病毒嵌合载体表达胶原酶Ⅱ能够有效地减轻大鼠肝纤维化程度.

AIM： To explore the therapeutic effect of an adenovirus-HBV chimeric vector expressing collagenase Ⅱ against cirrhosis in a rat model. METHODS： Rat liver cirrhosis was inducedwith 0.03% thioacetamide in drinking water for 16 wk. Ad-CH-tMMP8 and Ad-C-MMP8 were constructed using an adenovirus shuttle plasmid and a HBV chimeric vector expressing truncated and full-length collagenase Ⅱ gene. Ad-CHRFP2 expressing red fluorescent protein was used as a negative control. Liver cirrhosis rats were injected with the three plasmids through the tail vein. RESULTS： Compared with the model group and negative control group, fibrosis was dramatically attenuated four weeks after the infection. HE staining and picric acid-Sirius red. staining showed that hepatocyte steatosis, necrosis and inflammation were significantly milder in the treatment group, along with hepatocyte proliferation, recovery of hepatic lobule structure, and diminished content of HYP （28.97μg/g ± 2.36 μg/g vs 17.04 μg/g ± 0.61 μg/g, 17.62μg/g ± 1.30 μg/g, P 〈 0.05）, whereas the fibrosis in Ad-CHRFP2-treated rats persisted.CONCLUSION： Adenovirus-HBV chimeric vector expressing collagenase II effectively reduces the degree of liver fibrosis in rats.