木棉花总黄酮对CCl_4致肝纤维大鼠ColⅠ表达的影响

Impacts of the Total Flavone Glycoides( TFG) of Kapok on Col I Expression in the Rats of Liver Fibrosis Induced by CCl_4

目的研究木棉花总黄酮(TFG)对CCl4致肝纤维大鼠Ⅰ型胶原蛋白(ColⅠ)mRNA和ColⅠ蛋白表达的影响。方法采用50%CCl4花生油溶液灌胃诱导Wistar大鼠肝纤维化造模,将病理检查确认肝纤维化形成的Wistar大鼠50只随机分成5组,分别为模型对照组(模型组),秋水仙碱阳性对照组(阳性组),木棉花总黄酮高、中、低剂量组,另外设空白组,空白组和模型组给予等剂量的生理盐水,每日灌胃给药1次,连续4周。末次给药24 h后处死大鼠,RT-PCR检测肝组织中ColⅠmRNA的表达,用免疫组化方法观察肝组织ColⅠ蛋白质的表达,运用实时荧光定量PCR方法对肝组织ColⅠmRNA表达进行相对定量。结果 RT-PCR和实时荧光定量PCR结果均表明TFG能显著下调ColⅠmRNA的表达(P<0.05,P<0.01)。免疫组化结果显示,TFG能显著降低肝组织ColⅠ蛋白质的表达(P<0.05,P<0.01),且表现一定的剂量依赖性。结论 TFG对CCl4诱导的肝纤维大鼠具有一定的保护作用,其机制可能与其下调ColⅠmRNA及蛋白表达有关。

Objective To study the impacts of the total flavone glycoides （TFG） of Kapok on the ex- pressions of Col I mRNA and Col protein in the rats of liver fibrosis induced by CC14. Methods 50% CCl4 peanut oil solution was used for gastric infusion to induce the models of liver fibrosis in Wistar rats. Based on the identification of pathological examination ,50 cases of Wistar rats of liver fibrosis were randomized into 5 groups, named a model control group, colchicine （ Col ） positive control group, the high - dose TFG group, the middle - dose TFG group and the low - dose TFG group. Additionally, a blank control group was set up. The physiological saline solution of the same dose was used in the blank group and the model group via gastric in- fusion, once every day for 4 weeks continuously. The rats were sacrificed in 24 h after the last medication. RT - PCR was applied to detect the expression of Col I mRNA in liver tissue. IHC method was used to observe the expression of Col I protein in liver tissue. The real - time fluorescent quantitative PCR method was a- dopted for the relative quantitation of ColI mRNA expression in liver tissue. Results RT - PCR and the re- al - time fluorescent quantitative PCR methods all indicated that TGF down - regulated the expression of Col I mRNA significantly （P 〈 0.05,P 〈 0.01 ）. IHC results showed that TFG reduced significantly the expres- sion of Col I protein in liver tissue（P 〈 0.05, P 〈 0.01 ）, presenting a certain dose - dependence. Conclu- sion TFG presents a certain protective role in the rats of liver fibrosis induced by CCl4, which is probably related to its down - regulation of Col I mRNA and protein.