摘要
目的研究仲景炮附子Aconiti Lateralis Radix Preparata对慢性心力衰竭(CHF)的治疗作用,并采用超高效液相色谱四级杆静电场轨道阱高分辨质谱(UPLC-QE-Orbitrap-MS)测定仲景炮附子水煎液的成分,结合网络药理学探究其治疗CHF的作用机制。方法采用ip盐酸阿霉素构建大鼠CHF模型,ig给予仲景炮附子水煎液(3.3 g·kg^(-1))16d后,试剂盒法检测血清脑钠素(BNP)含量、心肌组织Ca^(2+)-ATP酶和Na^(+),K^(+)-ATP酶活性,苏木素-伊红(HE)染色后镜下观察左心室组织病理学形态变化。采用UPLC-QE-Orbitrap-MS测定仲景炮附子水煎液的成分,借助Swiss Target Prediction、DrugBank、Gene Cards、OMIM、TTD等平台预测仲景炮附子治疗CHF的作用靶点,利用Cytoscape软件构建相关作用网络,并对核心基因进行基因本体论(GO)功能富集分析和京都基因与基因组百科全书(KEGG)通路富集分析。结果与对照组比较,模型组大鼠出现稀便、腹水、体质量减轻等现象;血清BNP水平显著升高(P<0.01);Ca^(2+)-ATP酶、Na^(+),K^(+)-ATP酶活力均有降低趋势;左心室心肌组织切片出现明显的心肌纤维断裂,心肌间隙增宽,出现部分空泡变性等病理变化。与模型组比较,仲景炮附子水煎液组大鼠稀便情况得到一定程度改善,腹水和体型消瘦情况改善不明显;BNP水平显著降低(P<0.05);Ca^(2+)-ATP酶、Na^(+),K^(+)-ATP酶活力均有升高趋势;左心室组织病理学形态有明显改善。采用UPLC-QE-Orbitrap-MS共鉴定出仲景炮附子水煎液主要成分52个。通过网络药理学预测得到仲景炮附子治疗CHF活性成分43个,活性成分靶点472个,CHF疾病靶点1063个,活性成分与疾病交集靶点68个;网络分析结果表明,仲景炮附子可能通过作用于信号转导与转录激活子3(STAT3)、磷脂酰肌醇-4,5-二磷酸3-激酶催化亚基α(PIK3CA)、MAPK1等关键靶点发挥抗CHF作用,涉及到RAS相关蛋白1(Rap1)、磷脂酰肌醇3'-激酶(PI3K)-蛋白激酶B(Akt)、丝裂原活化蛋白激酶(MAPK)等信号通路。结论仲景炮附子对阿霉素诱导的慢CHF有一定的治疗作用,可通过多成分、多靶点、多途径发挥作用。
Objective To study the therapeutic effect of Zhongjing's Paofuzi on chronic heart failure(CHF),and use UPLC-QEOrbitrap-MS to determine the components of Zhongjing's Paofuzi decoction,combined with network pharmacology to explore its mechanism of action in the treatment of CHF.Methods The rat model of CHF was established by the ip of adriamycin hydrochloride.After 16 days of Zhongjing's Paofuzi decoction(3.3 g·kg^(−1)),the serum brain natriuretic peptide(BNP)level,the activities of Ca^(2+)-ATP and Na^(+),K^(+)-ATP enzymes in cardiac myocytes,and the cardiac histopathology were measured.UPLC-QEOrbitrap-MS was used to determine the components of Zhongjing's Paofuzi decoction,and the Swiss Target Prediction,DrugBank,Gene Cards,OMIM,and TTD platforms were used to predict the target of Zhongjing's Paofuzi in the treatment of CHF,and Cytoscape software was used to construct the correlation of effect network,and GO analysis and KEGG pathway enrichment for core genes were performed.Results Compared with the control group,the model group showed loose stool,ascites and weight loss,the serum BNP level was significantly increased(P<0.01),and the activities of Ca^(2+)-ATPase and Na^(+)-K^(+)-ATPase decreased.Besides,the myocardial tissue sections of the left ventricle showed obvious myocardial fiber rupture,widened myocardial space,and some pathological changes,such as vacuolar degeneration.Compared with model group,the loose stool of rats in Zhongjing's Paofuzi decoction group was improved to a certain extent,but the ascites and body weight loss were not improved significantly,the serum BNP level was decreased(P<0.05),and the activities of Ca^(2+)-ATP and Na^(+),K^(+)-ATP enzymes tended to increase.The histopathological morphology of left ventricle was significantly improved.A total of 52 main components of Zhongjing's Paofuzi decoction were identified by UPLC-QE-Orbitrap-MS.According to the network pharmacology prediction,43 active components,472 active component targets,1063 CHF disease targets and 68 intersection targets of active components and disease were obtained.The results of network analysis showed that Zhongjing's Paofuzi may exert its anti-heart failure effect by acting on key targets such as signal transduction and transcription activator 3(STAT3),Phosphatidylinositol-4,5-diphosphate 3-kinase catalytic subunit alpha(PIK3CA),mitogen-activated protein kinase 1(MAPK1),involving signaling pathways such as Ras-associated protein 1(Rap1),phosphatidylinositol 3'-kinase(PI3K)-Protein kinase B(Akt),and MAPK.Conclusion Zhongjing's Paofuzi has a certain therapeutic effect on CHF induced by adriamycin,and can treat CHF through the multi-component,multi-target and multi-way.
作者
焦鸣杰
李欣欣
刘淑明
陈倩
陈冬玲
尹贻慧
张凯
魏龙吟
张佳
李飞
JIAO Mingjie;LI Xinxin;LIU Shuming;CHEN Qian;CHEN Dongling;YIN Yihui;ZHANG Kai;WEI Longyin;ZHANG Jia;LI Fei(School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 102488,China)
出处
《药物评价研究》
CAS
2022年第10期1980-1991,共12页
Drug Evaluation Research
基金
国家自然科学基金面上项目(81973479)。
