黄芩-赤芍药对不同比例配伍抗大鼠肝纤维化模型作用机制探讨  

Mechanism of Scutellariae Radix-Paeoniae Radix Rubra Combination of Different Proportions Against Hepatic Fibrosis in Rats

作　　者：张敬博 陈平平[2] 于栋华[2] 刘苏杰 吴娟 刘树民[2] 王萍 ZHANG Jing-bo;CHEN Ping-ping;YU Dong-hua;LIU Su-jie;WU Juan;LIU Shu-min;WANG Ping(Heilongjiang University of Chinese Medicine,Harbin 150040,China;Institute of Traditional Chinese Medicine,Heilongjiang University of Chinese Medicine,Harbin 150040,China;Center for Drug Safety Evaluation,Heilongjiang University of Chinese Medicine,Harbin 150040,China;Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China)

机构地区：[1]黑龙江中医药大学,哈尔滨150040 [2]黑龙江中医药大学中医药研究院,哈尔滨150040 [3]黑龙江中医药大学药物安全性评价中心,哈尔滨150040 [4]中国中医科学院中药研究所,北京100700

出　　处：《中国实验方剂学杂志》2022年第12期69-77,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家重点基础研究发展计划(973计划)项目(2013CB531804);黑龙江省自然科学基金项目(LH2020H099);中国中医科学院科技创新工程项目(C12021A04904)。

摘　　要：目的:本研究旨在通过观察不同配伍比例的黄芩-赤芍对肝纤维化模型大鼠肝脏组织中Toll样受体4(TLR4)/髓样分化因子88(MyD88)/核转录因子-κB(NF-κB)和磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路表达水平的影响,探讨其干预肝纤维化的作用机制。方法:将60只SPF级雄性SD大鼠随机分正常、模型、水飞蓟素、黄芩-赤芍(1∶1)、黄芩-赤芍(1∶2)、黄芩-赤芍(1∶4)6组,每组10只。除正常组外,其余各组大鼠腹腔注射40%四氯化碳橄榄油溶液3 mL·kg^(-1)造模,首次5 mL·kg^(-1),每周2次,共计8周。后4周分别给予各组大鼠10 mL·kg^(-1)相应药物灌胃,并连续8周称量各组大鼠体质量。给药结束后,观察肝组织病理学变化,检测血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、透明质酸(HA)和层黏蛋白(LN)、白蛋白(ALB)、碱性磷酸酶(AKP)及肝组织中超氧化物歧化酶(SOD)活性,丙二醛(MDA)、羟脯氨酸(HYP)水平,实时荧光定量聚合酶链式反应(Real-time PCR)检测大鼠肝脏中TLR4、MyD88、NF-κB、PI3K、Akt、mTOR mRNA的表达水平。结果:与模型组比较,不同比例黄芩-赤芍均能回调各组大鼠体质量、改善肝脏病理损伤程度。酶联免疫吸附测定法(ELISA)结果显示,与正常组比较,模型组ALT、AST、HA、LN、AKP、MDA、HYP水平均有不同程度升高(P<0.05);与模型组比较,各给药组可明显降低ALT、AST、HA、LN、AKP、MDA、HYP含量及明显升高肝SOD活性、ALB水平(P<0.05)。Real-time PCR结果显示,与正常组比较,模型组TLR4、MyD88、NF-κB、PI3K、Akt、mTOR mRNA表达明显升高(P<0.05);与模型组比较,各给药组均能降低大鼠肝脏中TLR4、MyD88、NF-κB、PI3K、Akt、mTOR mRNA表达(P<0.05)。结论:不同比例黄芩-赤芍均可改善四氯化碳引起的肝组织病理学损伤,以黄芩-赤芍(1∶4)组疗效最佳。黄芩-赤芍可能通过抑制TLR4/MyD88/NF-κB和PI3K/Akt/mTObjective:To observe the effects of Scutellariae Radix(SR)-Paeoniae Radix Rubra(PRR combination of different proportions on the expression of the Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear transcription factorκB(NF-κB)and phosphatidylinositol kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR)signaling pathways in liver tissues of rats with hepatic fibrosis and explore the mechanism against hepatic fibrosis.Method:Sixty male SD rats of SPF grade were randomly divided into a normal group,a model group,a positive control(silymarin)group,and SR-PRR 1∶1,SR-PRR 1∶2,and SR-PRR 1∶4 groups,with 10 rats in each group.The hepatic fibrosis model was induced in rats except for those in the normal group by intraperitoneal injection of 40%tetrachloromethane(CCl4)-olive oil solution at 3 mL·kg^(-1),5 mL·kg^(-1)for the first time,for 8 weeks,twice per week.After 4 weeks,rats were treated correspondingly at 10 mL·kg^(-1)by intragastric administration,and the body weight of rats in each group was weighed for 8 weeks.After administration,histopathological changes in the liver were observed.Serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),hyaluronic acid(HA),laminin(LN),albumin(ALB),alkaline phosphatase(AKP),and superoxide dismutase(SOD)activities,malondialdehyde(MDA),and hydroxyproline(HYP)content in liver tissues were detected.The mRNA expression levels of TLR4,MyD88,NF-κB,PI3K,Akt,and mTOR in the liver of rats were detected by real-time fluorescencebased quantitative polymerase chain reaction(Real-time PCR).Result:Compared with the model group,SRPRR combination of different proportions could recover the body weight and improve the pathological injury of the liver.As revealed by enzyme linked immunosorbent assay(ELISA)results,compared with the normal group,the model group showed increased ALT,AST,HA,LN,AKP,MDA,and HYP levels to different degrees(P<0.05).Compared with the model group,the groups with drug intervention showed decreased levels of ALT,AS

关 键 词：黄芩-赤芍药对 肝纤维化 Toll样受体4(TLR4)/髓样分化因子88(MyD88)/核转录因子-κB(NF-κB) 磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/哺乳动物雷帕霉素靶蛋白(mTOR) 

分 类 号：R2-0[医药卫生—中医学] R22