基于非酒精性脂肪性肝炎模型及主成分分析的柴胡拆分组分引经药性归属探讨  

Meridian Tropism of Components in Bupleuri Radix Based on Nonalcoholic Steatohepatitis Model and Principal Component Analysis

作　　者：洪菲惠 陈洁欣 陈雨婵 李慧敏 彭东辉 沈志滨 夏永刚[1] 王秋红[1,2] 匡海学 HONG Feihui;CHEN Jiexin;CHEN Yuchan;LI Huimin;PENG Donghui;SHEN Zhibin;XIA Yonggang;WANG Qiuhong;KUANG Haixue(Key Laboratory of Basic and Application Research of Beiyao,Ministry of Education,Key Laboratory of Traditional Chinese Medicine(TCM)Pharmacodynamic Material Base,Heilongjiang University of Chinese Medicine,Harbin 150040,China;School of TCM,Guangdong Pharmaceutical University,Guangzhou 510006,China)

机构地区：[1]黑龙江中医药大学教育部北药基础与应用研究重点实验室,黑龙江中药及天然药物药效物质基础研究重点实验室,哈尔滨150040 [2]广东药科大学中药学院,广州510006

出　　处：《中国实验方剂学杂志》2022年第15期53-60,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家重点研发计划—中医药现代化研究项目(2019YFC1708801);国家重点研发计划项目(2018YFC1707100);广东省重点领域研发计划(2020B1111120002);黑龙江省“头雁”团队支持项目(黑龙江省头雁行动领导小组文件[2019]5号)。

摘　　要：目的:建立非酒精性脂肪性肝炎(NASH)模型,对逍遥散方剂配伍柴胡化学拆分组分,探究柴胡化学拆分归肝经作用强弱,并结合主成分分析明确柴胡归肝经药性物质基础。方法:采用80只SPF级雄性C57BL/6小鼠随机分为8组,每组10只,除正常组喂食蛋氨酸胆碱正常含量(MCS)饲料外,其余小鼠喂养蛋氨酸胆碱缺乏(MCD)模型饲料4周建立的NASH模型。苏木素-伊红(HE)染色证实模型建立后,分别灌胃给药4周,1次/d:逍遥散组2.874 g·kg^(-1)、逍遥散-柴胡组2.445 g·kg^(-1),逍遥散-柴胡+挥发油组、逍遥散-柴胡+多糖组、逍遥散-柴胡+黄酮组、逍遥散-柴胡+皂苷组于逍遥散-柴胡组基础上分别加入柴胡各拆分组分:挥发油0.163 mg·kg^(-1)、多糖24.067 mg·kg^(-1)、黄酮2.241 mg·kg^(-1)、皂苷2.746 mg·kg^(-1);模型组及正常组同体积生理盐水灌胃。末次灌胃给药后处死小鼠取血及肝脏组织,通过HE染色和油红O染色观察各组小鼠肝脏病理形态变化,酶联免疫吸附测定法(ELISA)试剂盒检测小鼠血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、甘油三酯(TG)、总胆固醇(TC)、高密度脂蛋白(HDL)、低密度脂蛋白(LDL)及肝脏丙二醛(MDA)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)及谷胱甘肽过氧化物酶(GSH-Px)水平考察其作用强弱;运用SPSS Statistics 23数据分析软件进行主成分分析,对逍遥散配伍柴胡不同拆分组分进行综合评价,确定柴胡归肝经药性物质基础。结果:与正常组比较,模型组小鼠肝细胞肿胀,脂肪空泡增多,有多处炎性细胞;血清ALT、AST、TG、TC、LDL含量升高,HDL含量降低(P<0.01);肝组织MDA含量升高,SOD、CAT及GSH-Px水平降低(P<0.01)。与模型组比较,逍遥散及其配伍柴胡不同拆分组分均可改善小鼠肝组织病理变化;各给药组血清ALT、AST水平均明显降低(P<0.05),逍遥散及其配伍柴胡不同拆分组分对小鼠血脂指标均有不同程度的改Objective: To explore the meridian tropism of components in Bupleuri Radix(Chaihu,CH)based on the model of nonalcoholic steatohepatitis(NASH)and clarify the substance basis of the meridian tropism of CH in Xiaoyaosan(XYS)by means of principal component analysis.Method: Eighty SPF male C57BL/6 mice were randomly assigned into 8 groups,with 10 mice in each group.Except that the blank group was fed with the methionine choline-sufficient(MCS)diet,the other mice were fed with methionine choline-deficient(MCD)diet for 4 weeks to establish the nonalcoholic steatohepatitis(NASH)model.After the established model was confirmed by hematoxylin-eosin(HE)staining,the mice were administrated with corresponding drugs by gavage once a day for 4 weeks.Specifically,the 8 groups were XYS group(2.874 g·kg^(-1)),XYS-CH group(2.445 g·kg^(-1)),XYS-CH+volatile oils(Vol,0.163 mg·kg^(-1))group,XYS-CH+polysaccharides(Pol,24.067 mg·kg^(-1))group,XYS-CH+flavones(Fla,2.241 mg·kg^(-1))group,and XYS-CH+saponins(Sap,2.746 mg·kg^(-1))group.The model group and the blank group were administrated with the same volume of normal saline.After the last administration,the mice were sacrificed for the collection of blood and liver tissue.The pathological changes of liver were observed by HE staining and oil red O staining.Enzyme linked immunosorbent assay(ELISA)kits were used to determine the levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),triglyceride(TG),total cholesterol(TC),high-density lipoprotein(HDL),and low-density lipoprotein(LDL)in serum as well as malondialdehyde(MDA),superoxide dismutase(SOD),catalase(CAT),and glutathione peroxidase(GSH-Px)in liver.SPSS Statistics 23 was used for principal component analysis and comprehensive evaluation to determine the substance basis of the meridian tropism of CH in NASH mice.Result: Compared with the blank control group,the modeling led to hepatocyte swelling,increased fat vacuoles,and appearance of inflammatory cells.Further,the modeling elevated the levels of ALT,AST,TG,TC,and

关 键 词：柴胡 化学拆分组分 逍遥散 归经药性理论 非酒精性脂肪性肝炎 主成分分析 

分 类 号：R2-0[医药卫生—中医学] R22R285.5R284R33