下瘀血汤抑制胰腺巨噬细胞浸润改善肝纤维化的机制研究  

Mechanism of Xiayuxue Decoction on improving liver fibrosis by inhibiting pancreatic macrophage infiltration

作　　者：沈东晓 马文婷 吴柳 张玮 陶乐 薛冬英[2] 张洁[2] 刘成[1,2] SHEN Dongxiao;MA Wenting;WU Liu;ZHANG Wei;TAO Le;XUE Dongyin;ZHANG Jie;LIU Cheng(Central Laboratory, Putuo Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 200062, China;Department of Infectious Diseases, Putuo Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai200062, China)

机构地区：[1]上海中医药大学附属普陀医院中心实验室,上海200062 [2]上海中医药大学附属普陀医院感染科,上海200062

出　　处：《上海中医药大学学报》2019年第2期66-72,79,共8页Academic Journal of Shanghai University of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(81673788);上海中医药大学中医学高峰学科项目(A-U151902);上海中医药大学预算内项目(2014YSN67)

摘　　要：目的:评价下瘀血汤对四氯化碳(CCl_4)诱导的肝纤维化模型小鼠肝脏和胰腺病变的改善作用,并探讨其相关机制。方法:24只雄性C57BL/6小鼠随机分为正常组、模型组和下瘀血汤组,每组8只。模型组和下瘀血汤组小鼠腹腔注射10%CCl_4诱导肝纤维化模型,正常组小鼠给予等体积橄榄油,每周3次,连续6周。造模第4周起,下瘀血汤组小鼠灌胃给予0.467 8 g/kg下瘀血汤药液,正常组和模型组小鼠给予等体积蒸馏水,每日1次,连续3周。药物干预期间,每周称量各组小鼠体质量。末次给药后,腹主动脉采血,分离肝、脾、胰腺组织并称重,计算肝脏指数和脾脏指数;试剂盒检测血清ALT、AST水平,HE染色和天狼星红染色后光镜下观察肝脏和胰腺组织病理形态学变化;免疫荧光检测肝组织α-SMA蛋白表达,免疫组化检测胰腺组织CD68、MCP-1及α-SMA蛋白表达。结果:①与正常组相比,模型组小鼠体质量明显降低(P<0.01),肝脏指数、脾脏指数显著增大(P<0.01),血清ALT、AST水平显著升高(P<0.05,P<0.01);与模型组相比,下瘀血汤组小鼠体质量显著升高,肝脏指数、脾脏指数及血清ALT、AST水平显著降低(P<0.01)。②病理学观察显示,模型组小鼠肝组织可见明显肝细胞肿胀、小叶中心性出血及灶状坏死,并伴有炎症细胞浸润,纤维组织大量增生,增生的胶原纤维分割肝小叶形成较粗大的间隔;胰腺小叶间隙增大,腺泡肿胀,腺管区有较多炎症细胞浸润。下瘀血汤组小鼠肝组织胶原纤维间隔较窄、排列疏松,肝细胞变性、坏死及肝和胰腺组织炎症细胞浸润明显减轻。③与正常组相比,模型组肝组织α-SMA表达明显增多,胰腺组织CD68、MCP-1及α-SMA阳性表达显著升高(P<0.01);下瘀血汤干预3周后,肝组织α-SMA表达明显减少,胰腺组织CD68、MCP-1及α-SMA阳性表达显著降低(P<0.01)。结论:肝纤维化可引起胰腺巨噬细胞浸润和星状细胞激活;下瘀血�Objective: To study the effects of Xiayuxue Decoction on the liver and pancreas lesions in mice with liver fibrosis induced by carbon tetrachloride(CCl4),and explore its related mechanisms. Methods: 24 male C57 BL/6 mice were divided into the normal group,modelgroup and Xiayuxue Decoction group,8 mice in each group. The mice in the model group and Xiayuxue Decoction group were treated with 10% CCl4 by intraperitoneal injection to induce liver fibrosis model,and the mice in the normal group were treated with olive oil at equal volume,3 times a week for 6 weeks. From the 4 th week of modeling,the Xiayuxue Decoction group was treated with Xiayuxue Decoction at dose of 0.467 8 g/kg,and the normal group and model group were treated with distilled water at equal volume,once every day for 3 weeks. During the drug intervention,the body mass of mice in each group was weighed weekly. After the last administration,the blood was collected from the abdominal aorta,and the liver,spleen and pancreas tissues were separated and weighed,then the indexes of liver and spleen were calculated. The serum levels of ALT and AST were detected by kits. The pathological changes of liver and pancreas were observed under light microscope after HE staining and Sirius red staining. The expression of α-SMA in liver tissue was detected by immunofluorescence,and the protein expressions of CD68,MCP-1 and α-SMA in pancreatic tissue were detected by immunohistochemistry.Results:(1)Compared with the normal group,the body weight of mice in the model group was significantly decreased( P<0.01),the liver index and the spleen index were significantly increased( P<0.01),and the serum levels of ALT and AST were remarkably increased( P<0.05,P<0.01). Compared with the model group,the body weight of mice in Xiayuxue Decoction group was obviously increased,and the liver index,the spleen index and the serum levels of ALT and AST were significantly decreased( P<0.01).(2)Pathological observation showed that hepatocyte swelling,central lobe hemorrhage and focal ne

关 键 词：下瘀血汤 肝纤维化 胰腺 巨噬细胞 小鼠 

分 类 号：R285.5[医药卫生—中药学]