藏药二十五味松石丸抑制血管生成和上皮间充质转化改善四氯化碳-酒精诱导的大鼠肝纤维化研究  

Tibetan medicine Ershiwuwei Songshi pills inhibit angiogenesis and epithelial mesenchymal transformation to improve carbon tetrachloride alcohol-induced liver fibrosis in rats

作　　者：许湘 黄丹[1,2] 陆华冠 刘延涛 刘馨 谢新 雷昌 赵洪庆[1,2] 刘建军 XU Xiang;HUANG Dan;LU Hua-guan;LIU Yan-tao;LIU Xin;XIE Xin;LEI Chang;ZHAO Hong-qing;LIU Jian-jun(Science and Technology Innovation Center,Hunan University of Traditional Chinese Medicine,Changsha 410208;State Key Laboratory Cultivation Base of Traditional Chinese Medicine Powder and Innovative Drugs,Changsha 410208;Department of Ultrasound,the Third Hospital of Sun Yat-Sen University Affiliated Hospital,Guangzhou 510630)

机构地区：[1]湖南中医药大学科技创新中心,长沙410208 [2]中药粉体与创新药物省部共建国家重点实验室培育基地,长沙410208 [3]中山大学附属医院第三医院超声科,广州510630

出　　处：《中南药学》2023年第5期1157-1163,共7页Central South Pharmacy

基　　金：湖南省教育厅资助科研项目(No.18A228);国家自然科学基金资助项目(No.82104836)。

摘　　要：目的研究二十五味松石丸是否通过抑制血管生成和上皮间充质转化改善四氯化碳(CCl_(4))-酒精诱导的大鼠肝纤维化模型。方法SPF级健康雄性SD大鼠30只,随机分为正常组,模型组,二十五味松石丸低、中和高[88、176、352 mg/(kg·d)]剂量组,每组6只。采用50%CCl_(4)橄榄油腹部皮下左右交替注射,同时以5%酒精饮料喂养建立肝纤维化模型;正常组皮下注射同体积橄榄油以蒸馏水喂养,持续8周,造模的同时按照分组灌胃相应的药物和0.4%羧甲基纤维素钠治疗。HE和Masson染色检测大鼠肝脏组织病理变化和胶原纤维增生;生化试剂盒测定血清谷丙转氨酶(ALT)、谷草转氨酶(AST),酶联免疫分析测定肿瘤坏死因子(TNF-α)、透明质酸(HA)和层黏连蛋白(LA)水平;免疫组化检测血小板内皮细胞黏附分子-1(CD31)、α-平滑肌肌动蛋白(α-SMA)、转化生长因子β1(TGF-β1)、E-钙黏蛋白(E-cadherin)和波形蛋白(Vimentin)表达;免疫印迹法检测α-SMA、TGF-β1、E-cadherin和Vimentin蛋白表达。结果与正常组相比,模型组大鼠肝组织病理损伤严重,胶原纤维明显增生;模型组中大鼠血清中ALT、AST活性明显升高,TNF-α含量明显增加,HA和LN含量明显升高;模型组大鼠肝组织内CD31、α-SMA、TGF-β1和Vimentin蛋白表达明显增加,E-cadherin蛋白表达明显降低。与模型组相比,二十五味松石丸各剂量组均能改善大鼠肝组织病理损伤、降低肝功能和肝纤维化指标,降低大鼠血清中TNF-α水平及肝组织内CD31、α-SMA、TGF-β1和Vimentin蛋白表达,增加E-cadherin蛋白表达。结论二十五味松石丸可改善CCl_(4)酒精复合因素诱导的大鼠肝纤维化,其作用可能与抑制血管新生和上皮间充质转化相关。Objective To determine whether Ershiwuwei Songshi pills improve Carbon tetrachloride(CCl_(4))alcohol-induced liver fibrosis in rats by inhibiting the angiogenesis and epithelial-mesenchymal transition.Methods Totally 30 SPF healthy male SD rats were randomly divided into a normal group,a model group,and a low,a middle,and a high dose group[88,176,and 352 mg/(kg·d)],with 6 rats in each group. The liver fibrosis model was established by subcutaneous injection of 50%CCl_(4) olive oil and feeding with 5% alcohol beverage. The normal group was fed with distilled water by subcutaneous injection of the same volume of olive oil for 8 weeks. Each group was treated with corresponding drugs and 0.4% sodium carboxymethylcellulose by gavage. HE staining and Masson staining were used to detect the pathological changes and collagen fiber proliferation in the rat liver. The levels of aspartate aminotransferase (AST), and alanine aminotransferase (ALT) in the serum were measured via biochemical kits;tumor necrosis factor-α (TNF-α ), hyaluronic acid (HA) and laminin (LA) in the serum were detected by ELISA. The expressios of platelet endothelial cell adhesion molecule-1 (CD31), α -smooth muscle actin (α -SMA), transforming growth factor-β 1 (TGF-β 1), E-cadherin and Vimentin were detected by immunohistochemistry. The protein expressions of α -SMA, TGF-β 1, E-cadherin and Vimentin were detected by Western blot. Results Compared with the normal group, the pathological damage in the liver tissue in the model group was severe, and the collagen fibers were obviously proliferated. The activities of ALT and AST in the serum of rats in the model group were significantly increased, the content of TNF-α was effectively increased, and the content of HA and LN was increased. The expression of CD31, α -SMA, TGF-β 1 and Vimentin protein in the liver tissue of the model group was effectively down-regulated, and the expression of E-cadherin protein was significantly decreased. Compared with the model group, all dose groups of Ershiwuw

关 键 词：二十五味松石丸 肝纤维化 血管生成 上皮间充质转化 血小板内皮细胞黏附分子-1 

分 类 号：R285[医药卫生—中药学]