基于代谢组学的四逆汤中附子-干姜药对配伍前后毒效变化规律分析  

Metabolomics-based Analysis of Toxicity Changes of Aconiti Lateralis Radix Praeparata-Zingiberis Rhizoma in Sinitang Before and After Compatibility

作　　者：彭伟[1,2] 刘冬梅 傅舒 王潇[1] 吴明权 何丕龙 涂禾 傅超美[1] PENG Wei;LIU Dongmei;FU Shu;WANG Xiao;WU Mingquan;HE Pilong;TU He;FU Chaomei(School of Pharmacy,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China;Sichuan Province Orthopedic Hospital,Chengdu 610041,China;Zigong First People's Hospital,Zigong 643000,China)

机构地区：[1]成都中医药大学药学院,成都611137 [2]四川省骨科医院,成都610041 [3]自贡市第一人民医院,四川自贡643000

出　　处：《中国实验方剂学杂志》2023年第18期126-136,共11页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81603536)。

摘　　要：目的:探讨附子干姜配伍前后的毒效变化规律,揭示二者配伍内涵。方法:将60只SD大鼠随机分为空白组、空白-附子组、空白-附子干姜组、模型组、模型-附子组、模型-附子干姜组,后3组采用尾静脉注射阿霉素复制心力衰竭大鼠模型,前3组尾静脉注射等量生理盐水,并对比分析附子单煎液与附子干姜合煎液对正常大鼠与模型大鼠生化指标、心肌组织病理学形态的影响;对大鼠血清样品进行代谢组学分析,运用主成分分析(PCA)和正交偏最小二乘法判别分析(OPLS-DA)筛选组间差异代谢物,并分析差异代谢通路;结合网络药理学筛选附子干姜配伍前后与增强抗心衰药效、调控心脏毒性相关的代谢物及其关联靶点、通路,采用蛋白免疫印迹法(Western blot)对筛选出的代表性通路进行验证。结果:与空白组比较,模型组脑钠肽(BNP)、肌酸激酶(CK)、乳酸脱氢酶(LDH)、肌钙蛋白(cTn)-T含量显著升高(P<0.01),空白-附子组CK、LDH、cTn-T含量明显升高(P<0.05,P<0.01),空白-附子干姜组CK含量显著升高(P<0.01);与空白-附子组比较,空白-附子干姜组LDH含量明显降低(P<0.05),病理切片显示2种煎液均可导致正常大鼠心肌组织病理损伤。与模型组比较,模型-附子干姜组BNP、CK、LDH、cTn-T含量明显降低(P<0.05,P<0.01),模型-附子组BNP、LDH、cTn-T含量明显降低(P<0.05,P<0.01);与模型-附子组比较,模型-附子干姜组CK含量显著降低(P<0.01),2种煎液均可改善模型大鼠心肌组织病理学形态。代谢组学结果显示,附子单煎液与附子干姜合煎液分别可回调模型大鼠422种与459种代谢物,附子干姜合煎液对正常大鼠的代谢干扰作用弱于附子单煎液。网络药理学关联分析结果显示,干姜增强附子抗心衰药效方面,脱氧尿苷酸等3个代谢物关联到56个代谢物,82个靶点,13条通路,包括钙信号通路、肾素分泌、肾素-血管紧张素系统等;干姜影响附子心�Objective:To explore the effect and toxicity change rule of Aconiti Lateralis Radix Praeparata(ALRP)and Zingiberis Rhizoma(ZR)before and after compatibility,and to reveal the compatibility connotation of them.Method:Sixty SD rats were randomly divided into blank group,blank-ALRP group,blank-ALRP-ZR group,model group,model-ALRP group and model-ALRP-ZR group,the latter three groups were injected with adriamycin via tail vein to establish the model of heart failure,and the former three groups were injected with the same amount of physiological saline via tail vein.The effects of ALRP single decoction and ALRP-ZR mixed decoction on biochemical indexes and myocardial histopathological morphology of normal rats and model rats were compared.Metabolomics analysis was performed on rat serum samples,principal component analysis(PCA)and orthogonal partial least squares-discriminant analysis(OPLS-DA)were used to screen the differential metabolites between groups,and the differential metabolic pathways were analyzed.Combined with network pharmacology technology,the metabolites and their associated targets and pathways related to enhancing anti-heart failure efficacy and reducing cardiotoxicity were screened before and after the compatibility of ALRP and ZR,the screened representative pathways were verified by Western blot.Result:Compared with the blank group,the model group showed significant increases in the contents of brain natriuretic peptide(BNP),creatine kinase(CK),lactate dehydrogenase(LDH)and cardiac troponin(cTn)-T(P<0.01),the blank-ALRP group showed obvious increases in CK,LDH,and cTn-T contents(P<0.05,P<0.01),while the normal-ALRP-ZR group showed a significant increase in CK conten(t P<0.01).Compared with the blank-ALRP group,the blank-ALRP-ZR group showed a obvious decrease in LDH content(P<0.05),and pathological sections showed that both decoctions could lead to myocardial histopathological damage in normal rats.Compared with the model group,the model-ALRP-ZR group showed obvious decreases in BNP,CK,LDH and cTn-T

关 键 词：附子 干姜 配伍 代谢组学 网络药理学 毒效变化规律 心力衰竭模型 

分 类 号：R22[医药卫生—中医基础理论] R969.1[医药卫生—中医学] R28[理学—分析化学] O657[理学—化学]