肝储脂细胞表达CYP11B2mRNA与实验性肝纤维化的关系研究

CYP11B2 mRNA exression in rat liver and its effect on CCL4induced-hepatic fibrogendsis of rat

目的比较CYP11B2在正常与实验性肝纤维化大鼠肝组织中的表达,评价安体舒通的抗纤维化作用。方法取雄性wistar大鼠160只,随机平均分为4组。模型组:CCLA油0.25mL/100mg皮下注射,每周三次;安舒体通组:CCLA油0.25ml/100mg皮下注射,每周三次;安体舒通20mgkg/d灌胃,1次/曰;马洛替脂组:CCLA油0.25ml/100mg皮下注射。每周三次:马洛替脂50ml/kg/d灌胃,1次/日;对照组:橄榄油0.25/ml/100mg皮下注射,每周次。于2、4、6、8、1O周末在光镜下动态观察组织学改变,图象分析仪测量胶原面积。RT-PCR和原位杂交检测纤维化及正常肝组织CYP11B2mRN的表达。结果 RT-PCR显示CYP11B2mRN在纤维化肝组织中表达上调。原位杂交显示CYP11B2mRNA表达定位于储脂细胞胞浆,在纤维化形成时表达增细。在肝纤维化形成的早中期阶段(6周以前),用安体舒通治疗能使肝纤维化分级和胶原面积减少。结论 CYP11R2mRNA在纤维化肝脏的储脂细胞中表达增强。安体舒通对早中期肝纤维化具有一定的治疗作用。

ob jective In consideration of the facts that extra-drenal tissues syntehesixe aldosteron not only in vascular tissue and brain but also in liver, and locally produced aldsterone is likely to take and active part in fibrogenesis of liver, we undertook present study to indentify aldosterone synthase bene-CYP11B2mRN Aexpression in normal and fibr- tic liver of rats and vevaluate the curative effect of antisterone. Methods: 120 Wistar rats weighting about 250g were divided into 4 groups. Modle group:The rats were injected with 40% CCLA0.25ml/100g subcutaneously three times a week. Antisterone group(n=30):The rats were injected with 40% CCLA0.25ml/100g subcutaneously three times a week. Antisterone equivalent to 20mg/kg/d was given ig. Malotilate grorp(n=30): The rats were injected with 40% CCLA 0.25mg/100g subcutaneously three times a week. Malotilate equivalent to 50mg/kg/d was given ig. Control group(n=30): the rats were injected with olive oil only. After2,4,6,8,and 10 weeks, the animals were sacrificed. Morphological examination was based on microscope and electron. The area and percentage of collagen were examined by Image Analyse system. By means of RT-PCR and in situ hybridization, the expressions of aldosterone synthase gene-CYP11B2 mRNA in fibrotic and normal liver were detected. Results In situ hybridization and RT-PCR showed the expression of CYP11B2mRNA, which localized in the endoplasm of fat storing cells (FSC_8), was up regulated when fibrogenesis occurred. Histological observation inndicated that the grade of fibrosis and the arade of collagen in antisterone group were less then thoae in model group before 6 weeks (P<0.05). There was no significant difference between antisterone group and malotilate group(P>0.05). After that, however, the grade of fibrosis and the area of collagen in antisterone group were higher than those in malotilate group(P<0.05). There was no significant difference between antisterone group and model group(P>0.05). Conclusion: The expression of CYP11B2 mRNA is up regulated in fibrotic liver. Antisterone can take fibrogenesis-inhibiting effect pratly in the early stage.