旋覆花汤对高脂高果糖高胆固醇饮食诱导非酒精性脂肪性肝炎小鼠模型的影响  

Effect of Xuanfuhua decoction on a mouse model of nonalcoholic steatohepatitis induced by high-fat,high-fructose,and high-cholesterol diet

作　　者：辛一敬 陈逸云 杨海琳 卓蕴慧[1] 张定棋 周峰峰 XIN Yijing;CHEN Yiyun;YANG Hailin;ZHUO Yunhui;ZHANG Dingqi;ZHOU Fengfeng(Department of Hepatology,Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Institute of Liver Diseases,Key Laboratory of Liver and Kidney Diseases,Ministry of Education,Shanghai Key Laboratory of Traditional Chinese Clinical Medicine,Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Department of Traditional Chinese Medicine,Shanghai Pudong New Area Gongli Hospital,Shanghai 200120,China;School of Pharmaceutical Sciences,School of TCM Research,Tsinghua University,Beijing 100084,China)

机构地区：[1]上海中医药大学附属曙光医院肝病科,上海201203 [2]上海中医药大学附属曙光医院肝病研究所,肝肾疾病病证教育部重点实验室,上海市中医临床重点实验室,上海201203 [3]上海市浦东新区公利医院中医科,上海200120 [4]清华大学药学院中药研究院,北京100084

出　　处：《临床肝胆病杂志》2023年第6期1340-1350,共11页Journal of Clinical Hepatology

基　　金：上海市科委科技支撑项目(19401934100);上海市科委科技计划项目(22Y11921000)。

摘　　要：目的探讨旋覆花汤对高脂高果糖高胆固醇饮食诱导小鼠非酒精性脂肪性肝炎(NASH)的干预作用。方法32只雄性C57/BL6J小鼠随机分为正常组、模型组、旋覆花汤组和奥贝胆酸组,每组8只。高脂高果糖高胆固醇饮食造模第24周开始,各组给予相应药物干预,旋覆花汤组给药剂量为14.19 g/kg灌胃,奥贝胆酸组给药剂量为10 mg/kg灌胃,灌胃体积均为20 mL/kg,每天给药1次,连续6周。HE染色、油红O染色、天狼星红染色及Masson染色观察肝组织病理学变化、脂质沉积及胶原沉积;试剂盒检测血清ALT、AST、TC、TG、LDL-C和葡萄糖水平,肝组织TG及羟脯氨酸(Hyp)含量;实时荧光定量聚合酶链式反应检测肝组织脂质代谢相关、炎症相关以及纤维化相关基因的表达;免疫组化染色观察肝组织F4/80和α-SMA阳性表达。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。结果与正常组相比,模型组小鼠体质量、肝湿重、血清AST、ALT、TC、TG、LDL-C和葡萄糖水平均显著增高(P值均<0.01);HE染色显示,模型组小鼠肝组织可见肝细胞脂肪变,大量脂肪空泡,肝细胞气球样变以及炎症细胞浸润,且非酒精性脂肪性肝病活动度评分(NAS)较正常组显著增加(P<0.01);油红O染色显示,模型组小鼠肝细胞内有大量红色脂滴沉积,大小不一,油红O染色面积百分比和肝脏TG含量较正常组均显著增加(P值均<0.01);天狼星红染色和Masson染色显示,模型组小鼠窦周、中央静脉以及汇管区胶原纤维增生明显,且肝组织Hyp含量较正常组显著增加(P<0.05)。与模型组比较,旋覆花汤组小鼠血清AST、ALT、TC、TG、LDL-C及葡萄糖水平均明显降低(P值均<0.05);肝组织脂肪变、炎症浸润、脂滴沉积以及胶原纤维增生明显改善,且NAS、油红O染色面积百分比、肝脏TG含量及Hyp含量均显著降低(P值均<0.05)。与正常组相比,模型组小鼠肝组织脂质代谢相关Objective To investigate the intervention effect of Xuanfuhua decoction on mice with nonalcoholic steatohepatitis(NASH)induced by high-fat,high-fructose,and high-cholesterol diet.Methods A total of 32 male C57/BL6J mice were randomly divided into normal group,model group,Xuanfuhua decoction group,and obeticholic acid group,with 8 mice in each group.Since week 24 of modeling using high-fat,high-fructose,and high-cholesterol diet,each group was given the corresponding drug for intervention at a dose of 14.19 g/kg by gavage for the Xuanfuhua decoction group and 10 mg/kg by gavage for the obeticholic acid group and a volume of 20 mL/kg for gavage,once a day for 6 consecutive weeks.HE staining,oil red O staining,Sirius Red staining,and Masson staining were used to observe the pathological changes,lipid deposition,and collagen deposition of liver tissue;related kits were used to measure the serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),total cholesterol(TC),triglyceride(TG),low-density lipoprotein cholesterol(LDL-C),and glucose,as well as the content of TG and hydroxyproline(Hyp)in liver tissue;quantitative real-time PCR was used to measure the expression of genes associated with lipid metabolism,inflammation,and fibrosis in liver tissue;immunohistochemical staining was used to observe the positive expression of F4/80 andα-SMA in liver tissue.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 Compared with the normal group,the model group had significant increases in body weight,liver wet weight,and serum levels of AST,ALT,TC,TG,LDL-C and glucose(all P<0.01).HE staining showed hepatocyte steatosis,a large number of fat vacuoles,hepatocyte ballooning degeneration,and inflammatory cell infiltration in liver tissue of the mice in the model group,and the model group had a significant increase in NAFLD activity score(NAS)compared with the norma

关 键 词：非酒精性脂肪性肝炎 旋覆花汤 小鼠 近交C57BL 

分 类 号：R285.5[医药卫生—中药学]