二甲双胍对布加综合征小鼠模型肝纤维化的影响  

Effect of metformin on liver fibrosis in a mouse model of Budd-Chiari syndrome

作　　者：杨净 李素新[1] 张跃辉[1] 李路豪 刘兆臣 沈东启 党晓卫[1] YANG Jing;LI Suxin;ZHANG Yuehui;LI Luhao;LIU Zhaochen;SHEN Dongqi;DANG Xiaowei(Department of Hepatobiliary and Pancreatic Surgery,The First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]郑州大学第一附属医院肝胆胰外科,郑州450052

出　　处：《临床肝胆病杂志》2022年第9期2034-2039,共6页Journal of Clinical Hepatology

基　　金：河南省医学科技攻关计划省部共建项目(SB201901003)。

摘　　要：目的探讨二甲双胍对于布加综合征小鼠模型肝脏纤维化的影响及其机制。方法C57雄性小鼠30只随机分为4组:假手术组(SHAM组)6只,假手术+二甲双胍组(SHAM+M组)5只,布加综合征模型组(BCS组)10只,布加综合征模型+二甲双胍组(BCS+M组)9只。模型组行下腔静脉部分结扎术,SHAM组不结扎,二甲双胍组造模同时饮水中给予0.1%二甲双胍。6周后处死小鼠取材,HE染色、天狼猩红染色观察小鼠肝脏组织病理和胶原沉积变化,免疫组化检测α-平滑肌肌动蛋白(α-SMA)、纤维蛋白原(Fibrinogen)表达情况,实时荧光定量PCR检测缺氧诱导因子-1α(HIF-1α)、Ⅰ型胶原(Collagen1)的mRNA表达,Western Blot检测HIF-1α、血管内皮生长因子(VEGF)、Fibrinogen、α-SMA、Collagen1蛋白的相对表达量。计量资料多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果与SHAM组相比,病理染色结果显示BCS组肝脏明显纤维化,中央静脉旁肝细胞排列紊乱,肝窦扩张并伴有红细胞淤积和少量炎症细胞浸润,胶原沉积。BCS组HIF-1α、Collagen1 mRNA表达增加,α-SMA、Collagen1、HIF-1α、VEGF、Fibrinogen蛋白表达明显增加(P值均<0.05);与BCS组相比,BCS+M组肝脏纤维化减轻,红细胞淤积减少,胶原沉积减少,HIF-1α、Collagen1 mRNA表达降低,α-SMA、Collagen1、HIF-1α、VEGF、Fibrinogen蛋白表达降低(P值均<0.05)。结论二甲双胍能够改善由布加综合征所导致的淤血性肝纤维化,其机制可能为减少肝窦内微血栓,抑制HIF-1α/VEGF通路来发挥作用。Objective To investigate the effect of metformin on liver fibrosis in a mouse model of Budd-Chiari syndrome and its mechanism.Methods A total of 30 male C57 mice were randomly divided into sham-operation group(SHAM group)with 6 mice,sham operation+metformin group(SHAM+M group)with 5 mice,Budd-Chiari model group(BCS group)with 10 mice,and Budd-Chiari model+metformin group(BCS+M group)with 9 mice.The mice in the model group were treated with partial ligation of the inferior vena cava,those in the SHAM group were not treated with ligation,and those in the metformin group were given 0.1% metformin in drinking water besides modeling.The mice were sacrificed after 6 weeks.HE staining and picrosirius red staining were used to observe liver histopathology and collagen deposition;immunohistochemistry was used to measure the expressions ofα-smooth muscle actin(α-SMA)and fibrinogen;quantitative real-time PCR was used to measure the mRNA expression of hypoxia-inducible factor 1α(HIF-1α)and type Ⅰ collagen(collagen 1),and Western blot was used to measure the relative protein expression levels of HIF-1α,vascular endothelial growth factor(VEGF),fibrinogen,α-SMA,and collagen 1.A one-way analysis of variance was used for comparison of continuous data between multiple groups,and the least significant difference t-test was used for further comparison between two groups.Results Pathological staining showed that compared with the SHAM group,the BCS group had significant liver fibrosis,disordered arrangement of hepatocytes near the central vein,sinusoidal expansion with red blood cell deposition and a small amount of inflammatory cell infiltration,and collagen deposition.The BCS group had significant increases in the mRNA expression levels of HIF-1α and collagen 1 and the protein expression levels of α-SMA,collagen 1,HIF-1α,VEGF,and fibrinogen(all P<0.05);compared with the BCS group,the BCS+M group had significant alleviation of liver fibrosis,red blood cell deposition,and collagen deposition and significant reductions in t

关 键 词：BUDD-CHIARI综合征 肝硬化 二甲双胍 小鼠 近交C57BL 

分 类 号：R575.2[医药卫生—消化系统]