摘要
目的初步探索扶正养肝化纤汤对CCl_(4)诱导肝纤维化小鼠的治疗作用及其机制。方法取6周龄19~22 g的SPF级雄性C57BL/6J小鼠48只,根据随机数字表法将其分为空白组,模型组,扶正养肝化纤汤高、中、低剂量组,水飞蓟素治疗组,每组8只。除空白组外,其余5组予5 ml/kg橄榄油+20%CCl_(4)腹腔注射,3次/周,持续8周。模型构建的同期各组给予相应的药物灌胃治疗,1次/d,连续8周。观察小鼠一般状态变化;比较各组肝脏指数,血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、基质金属蛋白酶-1(MMP-1)、金属蛋白酶组织抑制剂-1(TIMP-1)水平;比较各组肝组织MMP-1、TIMP-1 mRNA水平;苏木精-伊红染色观察肝组织病理学变化;免疫荧光染色观察肝脏星状细胞(HSC)变化。结果空白组状态良好;模型组精神萎靡,活动迟缓,皮毛欠光泽,食欲减退;扶正养肝化纤汤高、中、低剂量组毛发干枯稀疏情况好于模型组,饮食、饮水较模型组稍多,活动尚可。造模第3周开始,模型组体重低于空白组;扶正养肝化纤汤高、中、低剂量组体重高于模型组,且扶正养肝化纤汤高剂量组体重下降趋势不明显。模型组肝脏指数,血清ALT、AST水平高于空白组(P<0.05)。扶正养肝化纤汤高、中、低剂量组肝脏指数,血清ALT、AST水平低于模型组(P<0.05);扶正养肝化纤汤高、中剂量组肝脏指数,血清ALT、AST水平低于扶正养肝化纤汤低剂量组;扶正养肝化纤汤高剂量组肝脏指数,血清ALT、AST水平低于扶正养肝化纤汤中剂量组(P<0.05)。模型组血清MMP-1水平、肝组织MMP-1 mRNA水平低于空白组,血清TIMP-1水平、肝组织TIMP-1 mRNA水平高于空白组(P<0.05)。扶正养肝化纤汤高、中、低剂量组血清MMP-1水平、肝组织MMP-1 mRNA水平高于模型组,血清TIMP-1水平、肝组织TIMP-1 mRNA水平低于模型组(P<0.05)。扶正养肝化纤汤高、中剂量组血清MMP-1水平、肝组织MMP-1 mRNA水平高于扶正养肝化纤汤低剂量组,血清TIMP-1水平、肝组织TIMP-1 mRNA水平低于扶正养肝化纤汤低剂量组;扶正养肝化纤汤高剂量组血清MMP-1水平、肝组织MMP-1 mRNA水平高于扶正养肝化纤汤中剂量组;血清TIMP-1水平、肝组织TIMP-1 mRNA水平低于扶正养肝化纤汤中剂量组(P<0.05)。空白组肝小叶结构边界清晰完整,细胞形态正常;模型组肝脏形态损伤、炎症细胞浸润,肝脏HSC激活数量高于空白组;扶正养肝化纤汤高、中、低剂量组肝脏形态损伤、炎症细胞浸润,肝脏HSC激活数量低于模型组。结论高剂量扶正养肝化纤汤治疗肝纤维化可明显减少HSC激活数量,降低细胞外基质沉积,改善肝组织病理,其作用机制之一可能与调控肝组织MMPs/TIMPs活性平衡有关。
Objective To explore the therapeutic effect and me-chanism of Fuzheng Yanggan Huaxian Decoction on CCl_(4)-induced liver fibrosis in mice.Methods A total of 48 SPF grade male C57BL/6J mice aged 6 weeks and weighing 19-22 g were divided into blank group,model group,high-,medium-,and low-dose groups of Fuzheng Yanggan Huaxian Decoction,and Silymarin treatment group according to random number table method,with eight mice in each group.In addition to blank group,the remaining five groups were given 5 ml/kg olive oil+20%CCl_(4)intrabitoneal injection,three times a week,for eight weeks.In the same period of the model construction,each group was given corresponding drug intragastric therapy,once a day,for eight weeks.The general state of mice was observed;liver index,the levels of serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),matrix metalloproteinase-1(MMP-1),and tissue inhibitor of metalloproteinase-1(TIMP-1)were compared among all groups;the levels of MMP-1 and TIMP-1 mRNA in liver tissues of all groups were compared;hematoxylin-eosin staining was used to observe the histopathological changes of liver;and the changes of hepatic stellate cells(HSC)were observed by immunofluorescence staining.Results The blank group was in good condition;in model group,the spirit was weak,the activity was slow,the fur was not shiny,and the appetite was decreased;the dry and sparse hair in high-,medium-,and low-dose groups of Fuzheng Yanggan Huayxian Decoction was better than that of model group,and the diet and drinking water were slightly more than that of model group.The liver index and the levels of serum AST and ALT of model group were higher than those of blank group(P<0.05).The liver index and the levels of serum AST and ALT in high-,medium-,and low-dose groups of Fuzheng Yanggan Huayxian Decoction were lower than those of model group(P<0.05).The liver index and the levels of serum AST and ALT in high-and medium-dose groups of Fuzheng Yanggan Huayxian Decoction were lower than those in low-dose group of Fuzheng Yanggan Huayxian Decoction;and the liver index and the levels of serum AST and ALT in high-dose group of Fuzheng Yanggan Huayxian Decoction were lower than those in medium-dose group of Fuzheng Yanggan Huayxian Decoction(P<0.05).The levels of serum MMP-1 and liver tissue MMP-1 mRNA in model group were lower than those in blank group,while the levels of serum TIMP-1 and liver tissue TIMP-1 mRNA were higher than those in blank group(P<0.05).The levels of serum MMP-1 and liver tissue MMP-1 mRNA in high-,medium-,and low-dose groups of Fuzheng Yanggan Huayxian Decoction were higher than those in model group,while the levels of serum TIMP-1 and liver tissue TIMP-1 mRNA were lower than those in model group(P<0.05).The levels of serum MMP-1 and liver tissue MMP-1 mRNA in high-and medium-dose groups of Fuzheng Yanggan Huayxian Decoction were higher than those of low-dose group of Fuzheng Yanggan Huayxian Decoction;while the levels of serum TIMP-1 and liver tissue TIMP-1 mRNA were lower than those in low-dose group of Fuzheng Yanggan Huayxian Decoction(P<0.05).The levels of serum MMP-1 and liver tissue MMP-1 mRNA in high-dose group of Fuzheng Yanggan Huayxian Decoction were higher than those in medium-dose group of Fuzheng Yanggan Huayxian Decoction;while the levels of serum TIMP-1 and liver tissue TIMP-1 mRNA were lower than those in medium-dose group of Fuzheng Yanggan Huayxian Decoction(P<0.05).In blank group,the boundary of hepatic lobular structure was clear and complete,and the cell morphology was normal;in model group,liver morphology was damaged and inflammatory cells infiltrated,and the number of hepatic HSC activation was higher than that of blank group;and in high-,medium-,and low-dose groups of Fuzheng Yanggan Huayxian Decoction,the liver morphological damage,inflammatory cell infiltration,and the number of hepatic HSC activation was lower than that of model group.Conclusion In the treatment of liver fibrosis,high-dose Fuzheng Yanggan Huaxian Decoction can significantly reduce the number of HSC activation,decrease extracellular matrix deposition,and improve liver histopathology.One of its mechanisms may be related to the regulation of MMPs/TIMPs activity balance in liver tissue.
作者
陈欣
朱婷
臧雪
赵鑫
栾云鹏
李襄
王华宁
CHEN Xin;ZHU Ting;ZANG Xue;ZHAO Xin;LUAN Yunpeng;LI Xiang;WANG Huaning(The First Clinical College,Yunnan University of Chinese Medicine,Yunnan Province,Kunming650021,China;Department of Spleen and Stomach Diseases,the First Affiliated Hospital of Yunnan University of Chinese Medicine,Yunnan Province,Kunming650500,China)
出处
《中国医药导报》
CAS
2024年第33期13-19,共7页
China Medical Herald
基金
云南省科技厅中医联合专项-基础研究计划项目(202301AZ070001-039)
云南省科技厅中医联合专项-基础研究专项项目(202301AT070251)
云南省教育厅科学研究基金项目(2023Y0475)。
关键词
肝纤维化
扶正养肝化纤汤
细胞外基质
基质金属蛋白酶
金属蛋白酶组织抑制剂
Liver fibrosis
Fuzheng Yanggan Huaxian Decoction
Extracellular matrix
Matrix metalloproteinases
Tissue inhibitor of metalloproteinase
作者简介
陈欣(1998.2-),女,云南中医药大学第一临床医学院2022级中医内科学专业在读硕士研究生,研究方向:中医药防治脾胃肝胆病。;通讯作者:王华宁(1966.4-),女,硕士,主任医师,教授,研究方向:中西医防治脾胃肝胆病。
