Inhibitory effect of Anfate on expression of TGFβ1 /Smad2 signaling pathway in treatment of hepatic fibrosis in mice
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摘要:
目的研究复方牛胎肝提取物通过抑制肝纤维化小鼠肝脏组织转化生长因子β1/Smad 2通路发挥抗纤维化作用的机制。方法健康雄性ICR小鼠90只,随机分为正常对照组(10只),模型组(20只),治疗组(60只)。治疗组按不同剂量分为大剂量治疗组(20只),中剂量治疗组(20只),小剂量治疗组(20只)。模型组及治疗组皮下注射30%四氯化碳溶液制备肝纤维化小鼠模型。治疗组造模8周后用复方牛胎肝提取物以大、中、小剂量灌胃治疗,1次/d,持续4周;正常组及模型组同期予等量生理盐水灌胃。肝脏组织经HE染色后光学显微镜观察病理变化;采用Real-Time PCR、免疫组织化学及Western蛋白印迹法分别检测正常组、模型组及治疗组TGFβ1、蠕虫果蝇母抗同源蛋白(Smad)2的mRNA及蛋白表达水平。各组间均数比较采用单因素方差分析。方差齐性采用Levene法检验,样本均数比较采用单因素方差分析;方差不齐时采用秩和检验。相关性检验采用Pearson直线相关分析。结果正常对照组TGFβ1、Smad 2 mRNA及蛋白表达量很低,模型组较高,治疗组较模型组表达下降,大剂量治疗组下降最为显著,其他2组随治疗...
Abstract:Objective To investigate the inhibitory effect of Anfate on the expression of transforming growth factor β1 (TGFβ1) /small mothers against decapentaplegic homolog 2 (Smad2) signaling pathway in the treatment of hepatic fibrosis in mice.Methods Ninety healthy male ICR mice were randomly divided into three groups:normal control group (n = 10) , model group (n = 20) , and high-, middle-, and low- dose treatment groups (n = 20 for each) .In the model group and treatment groups, a mouse model of hepatic fibrosis was established by hypodermic injection of 30% carbon tetrachloride (CCl4) .At 8 weeks after the model was established, the mice in treatment groups received high-, middle-, or low- dose Anfate by gavage once daily for 4 weeks, while the normal control group and model group received the same volume of saline by gavage.The liver tissues were observed under a light microscope after HE staining.The mRNA and protein expression levels of TGFβ1 and Smad2 were measured by Real- Time PCR and immunohistochemistry / Western Blot, respectively.Comparison of means between groups was made by One- Way analysis of variance (ANOVA) .Levene's test was used for assessing the equality of variances;comparison of sample means was made by One- Way ANOVA, and the Kruskal- Wallis test was used when the variances were unequal.Correlation analysis was performed using the Pearson linear correlation coefficient.Results The mRNA and protein expression of TGFβ1 and Smad2 was low in the normal control group and high in the model group;compared with the model group, the treatment groups showed decreases in the mRNA and protein expression of TGFβ1 and Smad2, most significant in the high- dose treatment group, followed by the middle- dose treatment group and high- dose treatment group;there were significant differences in the mRNA expression of TGFβ1 and Smad2 between these groups (χ2= 27.877, P < 0.05;χ2= 26.688, P < 0.05) .The linear correlation analysis showed that the mRNA expression of TGFβ1 and Smad2 was negatively correlated with the dose of Anfate (r =- 0.967, P < 0.05;r =- 0.956;P < 0.05) .Conclusion Anfate inhibits the expression of TGFβ1 and Smad2 in liver tissues in a dose- dependent manner, and that may be related to the mechanism by which Anfate relieves the hepatic fibrosis in mice.
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Key words:
- liver cirrhosis /
- liver extracts /
- transforming growth factor beta /
- Smad2 protein /
- mice
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