双氢青蒿素抑制四氯化碳诱导的小鼠肝纤维化的实验研究

The inhibitor effect of dihydroartemisinin on liver fibrosis of mice induced by CCl_4

目的探讨双氢青蒿素(DHA)对CCl_4诱导的小鼠肝纤维化的抑制作用及其机制。方法 2013年10月—2014年5月河北工程大学医学院中心实验室进行实验,将50只小鼠分为3组:对照组10只、模型组20只和治疗组20只。模型组与治疗组采用四氯化碳(CCl_4)腹腔注射的方法制备小鼠肝纤维化模型。治疗组小鼠按20μg/g体质量的DHA灌胃,每日1次,共12次;对照组、模型组小鼠给予等剂量的DMSO生理盐水灌胃,每日1次,共12次。HE和Masson三色染色法观察各组肝组织病理学变化;比色法测定肝组织中的羟脯氨酸的含量;Western blot方法检测基质金属蛋白13(MMP-13)及其组织抑制因子1(TIMP-1)的表达水平。结果 HE和Masson三色染色结果证实小鼠肝纤维化动物模型的成功建立。模型组小鼠肝内出现肝细胞脂肪变性,纤维组织明显增生,肝组织中MMP-13的表达降低,而羟脯氨酸含量和TIMP-1表达增加,MMP-13/TIMP-1值下降,与对照组相比差异均有统计学意义(P<0.05)。治疗组经过DHA治疗后,小鼠肝内纤维组织增生降低,肝组织中MMP-13的表达增加,而羟脯氨酸含量和TIMP-1表达降低,与模型组相比差异均有统计学意义(P<0.05)。结论双氢青蒿素具有抗肝组织纤维化作用,提示可作为一种新型抗肝组织纤维化药物。

Objective To investigate the inhibitory effect of（DHA） on hepatic fibrosis induced by CCl_4 in mice and its mechanism.Methods From October 2013 to May 2014,in the Central Laboratory of Hebei University of Engineering Medical College,50 mice were divided into 3 groups：control group（n = 10）,model group（n =20） and treatment group（n = 20）.Model group and treatment group were treated with intraperitoneal injection of carbon tetrachloride（CCl_4） to prepare the model of liver fibrosis mice.Mice in the treatment group were administered with 20 μg/g,1 times a day,12 times,the control group and the model group were given DMSO normal saline for 1 times a day,12 times.HE and Massons trichrorae staining were used to observe the pathological changes of liver tissue;colorimetric method were used for the determination of the content of hydroxyproline in liver tissue;Western blot method were used for the detection of matrix metalloproteinase 13（MMP-13） tissues and the expression level of inhibit factor 1（TIMP-1）.Results The HE and Masson staining results confirmed the successful establishment of animal model of liver fibrosis in mice.In model group mice＇ s liver,fatty degeneration of liver cell were found,marked hyperplasia of fibrous tissue,reduce the liver tissue in MMP-13 expression,and hydroxyproline content and TIMP-1 expression increased,and MMP-13/TIMP-1 value were decreased than the control group（P〈0.05）.Treatment group after DHA therapy,mice liver fibrous tissue hyperplasia decreased,increased liver tissue＇s MMP-13 expression,and hydroxyproline content and TIMP-1 expression decreased than the model group（P〈0.05）.Conclusion Dihydroartemisinin has the function of anti-hepatic fibrosis,which may be as a new type of anti-hepatic fibrosis drug.