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.
DownLoad: