氧化苦参碱对免疫性肝纤维化的胶原纤维合成和TGF-β_1mRNA的影响

目的:研究氧化苦参碱对小鼠免疫性肝纤维化胶原纤维合成和TGF-β_1 mRNA的影响。方法:将BABL/c小鼠随机分成正常对照组、阳性对照组、氧化苦参碱小剂量、大剂量治疗组,阳性对照组和氧化苦参碱治疗组每周静脉注射1次刀豆蛋白A,共20W。氧化苦参碱治疗组分别腹腔注射氧化苦参碱30mg/kg、60mg/kg。共20W,分别于注射刀豆蛋白A后5、12和20W取血低温保存,并每次处死1批小鼠取肝脏组织液氮低温保存,常规HE染色、VanGieson胶原纤维染色,图像分析系统定量分析肝内纤维组织含量。抽提小鼠肝脏组织总RNA,RT—PCR方法扩增检测肝组织内TGF-β_1、β-actin。凝胶图像分析系统扫描扩增产物的灰度值,以β-actin作为内参照,计算相对表达量。结果:氧化苦参碱治疗组血清ALT含量及肝组织内炎症活动度、纤维含量均低于阳性对照组,而且呈剂量依赖性,TGF-β_1 mRNA相对表达量下降。结论:氧化苦参碱有减轻小鼠肝脏炎症活动度并可能通过下调TGF-β_1的表达而抑制小鼠免疫性肝纤维化的发生。

Effect of Anluohuaxian Tablet Combined with γ-IFN on Schistosomal Liver Fibrosis

The therapeutic effects of anluohuaxian tablet combined with γ-IFN on schistosomal liver fibrosis and its mechanism were studied in a murine model and clinical cases of schistosomal liver fibrosis, Fifty Kunming mice were randomly divided into 5 groups： normal control group, infection control group, anluohuaxian tablet-treated group, γ-IFN-treated group and combined treatment （anluohuaian tablet＋γ-IFN） group. Pathologic changes in liver, including hepatic pigmentation and the size of schistosomal egg granuloma, were observed by HE staining after treatment for 8 weeks. The expression of the type Ⅰ and Ⅲ collagen, and TIMP-1 was detected by immunohistochemistry. TGF-β1 mRNA expression was examined by real-time fluorescent quantitative PCR. Sixty patients with schistosomal liver fibrosis were divided into treatment group and control group. The patients in treatment group were treated with anluohuaxian tablet in combination with γ-IFN for 6 months. Before and after treatment, the changes of symptoms and signs, liver function, serum liver fibrosis indexes and imaging indexes were observed. The results showed that as compared with infection control group, all forms of treatments relieved the hepatic pathological injury with apparently diminished size of schistosomal egg nodules and decreased percentage of pigmentation （P〈0.05）. Furthermore, the expression of collagen Ⅰ and Ⅲ, TIMP-1, and TGF-β1 mRNA in combined treatment group was significantly decreased as compared with anluohuaxian tablet-treated and γ-IFN-treated groups （P〈0.05）. In the clinical observation, the serum liver fibrosis indexes, the portal vein width as well as the spleen thickness was significantly reduced in treatment group as compared with control group （P〈0.05）. It was concluded that the combined use of anluohuaxian tablet with γ-IFN in schistosomal liver fibrosis could protect liver function, alleviate liver fibrosis, and could be used as a choice in treating patients with schiatosomal liver fibrosis.

安络化纤丸联合γ-干扰素对血吸虫肝纤维化鼠TIMP-1及TGF-β_1的影响

目的了解安络化纤丸+γ-干扰素(IFN-γ)对实验性血吸虫病肝纤维化指标TIMP-1、TGF-β1表达的影响。方法将36只昆明小鼠随机分为正常对照组,感染对照组及IFN-γ+安络化纤丸组;采用日本血吸虫尾蚴40条/只鼠攻击感染小鼠建立血吸虫性肝纤维化模型,IFN-γ+安络化纤丸联合干预连续8周,观察肝组织纤维化程度及虫卵结节变化;SABC免疫组织化学检测TIMP-1的表达;实时荧光定量法检测TGF-β1mRNA的含量。结果安络化纤丸+IFN-γ组小鼠肝组织虫卵肉芽肿个数较感染对照组少,虫卵肉芽肿平均直径较感染对照组小,两组相比,差异有统计学意义(P<0.05);安络化纤丸+IFN-γ组小鼠肝组织TIMP-1的水平和TGF-β1mRNA的相对量均较感染对照组显著下降,差异有统计学意义(P<0.05)。结论安络化纤丸+IFN-γ组虫卵肉芽肿减小,数目减少,肝纤维化程度减轻可能与其能减少TIMP-1的表达,下调TGF-β1的转录水平有关。

抗纤Ⅰ号等方剂和硒抗大鼠肝纤维化作用的实验研究

目的 :研究抗纤Ⅰ号、抗纤Ⅱ号、抗纤Ⅲ号及各方剂加硒对大鼠肝纤维化治疗作用和免疫调节作用 ,并从分子水平对其机制进行探讨。方法 :利用四氯化碳、胆固醇等复合因素制造大鼠肝纤维化模型 ,设立空白对照组、肝纤维化模型组、模型自然恢复组、西药组及中药治疗组 ,中药灌胃治疗 4 2d ,检测病理学指标、血清酶学指标、血清学肝纤维化指标、免疫学指标 ,以及肝组织TGF β1mRNA表达量的变化。结果 :各项检查均显示抗纤Ⅰ号具有很好的抗肝纤维化的作用 ,其疗效优于抗纤Ⅱ号和抗纤Ⅲ号 ,抗纤Ⅱ号优于抗纤Ⅲ号 ,因此推测丹参为抗纤Ⅰ号的主药 ,鳖甲和郁金的辅助作用优于三棱和莪术。中药治疗组大鼠胸腺组织CD4、CD4/CD8均明显高于秋水仙碱组。加硒组与各方剂相比大鼠胸腺组织CD4、CD4/CD8均有升高 ,但差异无显著性。同时实验表明抗纤Ⅰ号能够明显抑制TGF β1mRNA的表达量。结论 :抗纤Ⅰ号对大鼠肝纤维化具有明确的治疗作用 ,且有提高机体免疫力的作用。其作用机制可能是通过下调TGF β1mRNA 。