CAJ | 학술논문

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组随治疗剂量的减少表达逐渐增加；各组TGFβ1、Smad 2 mRNA表达水平差异具有统计学意义（χ2＝27．877、26．688，P＜0．05）。直线相关性分析显示：各治疗组TGFβ1、Smad 2的mRNA表达水平与治疗剂量均呈负相关（r＝-0．967、-0．956，P＜0.05）。结论复方牛胎肝提取物能改善小鼠肝脏纤维化程度，并呈剂量依赖性抑制TGFβ1、Smad 2蛋白在肝纤维化组织中的表达，这可能与其抗肝纤维化机制相关。
목적：연구복방우태간제취물통과억제간섬유화소서간장조직전화생장인자β1/Smad 2통로발휘항섬유화작용적궤제。방법건강웅성ICR소서90지，수궤분위정상대조조（10지），모형조（20지），치료조（60지）。치료조안불동제량분위대제량치료조（20지），중제량치료조（20지），소제량치료조（20지）。모형조급치료조피하주사30％사록화탄용액제비간섬유화소서모형。치료조조모8주후용복방우태간제취물이대、중、소제량관위치료，1차/d，지속4주；정상조급모형조동기여등량생리염수관위。간장조직경HE염색후광학현미경관찰병리변화；채용Real-Time PCR、면역조직화학급Western 단백인적법분별검측정상조、모형조급치료조TGFβ1、연충과승모항동원단백（Smad）2적mRNA급단백표체수평。각조간균수비교채용단인소방차분석。방차제성채용Levene법검험，양본균수비교채용단인소방차분석；방차불제시채용질화검험。상관성검험채용Pearson직선상관분석。결과정상대조조TGFβ1、Smad 2 mRNA급단백표체량흔저，모형조교고，치료조교모형조표체하강，대제량치료조하강최위현저，기타2조수치료제량적감소표체축점증가；각조TGFβ1、Smad 2 mRNA표체수평차이구유통계학의의（χ2＝27．877、26．688，P＜0．05）。직선상관성분석현시：각치료조TGFβ1、Smad 2적mRNA표체수평여치료제량균정부상관（r＝-0．967、-0．956，P＜0.05）。결론복방우태간제취물능개선소서간장섬유화정도，병정제량의뢰성억제TGFβ1、Smad 2단백재간섬유화조직중적표체，저가능여기항간섬유화궤제상관。
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-,mid-dle-,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 e-quality of variances;comparison of sample means was made by One-Way ANOVA,and the Kruskal-Wallis test was used when the vari-ances were unequal.Correlation analysis was performed using the Pearson linear correlation coefficient.Results The mRNA and protein ex-pression of TGFβ1 and Smad2 was low in the normal control group and high in the model group;compared with the model group,the treat-ment 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.