基于UPLC-Q-TOF/MS整合网络药理学探讨平肝益肾方治疗自发性高血压大鼠靶器官损害的效应机制  

Exploring the Mechanism of Pinggan Yishen Decoction Against Target Organ Damage in Spontaneously Hypertensive Rats Based on UPLC-Q-TOF/MS and Network Pharmacology

作　　者：章伟婷 王琼[1,4] 樊亚东 王慧惠[1,2,3] 陈山珊 张思奇[1,2,3] 陈溢滢 吴磊[1] 戴国梁[1] 宋冰冰 方祝元[1,2,3] ZHANG Weiting;WANG Qiong;FAN Yadong;WANG Huihui;CHEN Shanshan;ZHANG Siqi;CHEN Yiying;WU Lei;DAI Guoliang;SONG Bingbing;FANG Zhuyuan(Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029,China;Institute of Hypertension,Jiangsu Provincial Hospital of Traditional Chinese Medicine,Nanjing 210001,China;Clinical Medical Innovation Center for Hypertension,Jiangsu Provincial Hospital of Traditional Chinese Medicine,Nanjing 210019,China;Basic Pharmacology Laboratory,Jiangsu Provincial Hospital of Traditional Chinese Medicine,Nanjing 210029,China)

机构地区：[1]南京中医药大学附属医院,江苏南京210029 [2]江苏省中医院高血压研究所,江苏南京210001 [3]江苏省中医院高血压病临床医学创新中心,江苏南京210019 [4]江苏省中医院基础药理实验室,江苏南京210029

出　　处：《南京中医药大学学报》2024年第9期949-961,共13页Journal of Nanjing University of Traditional Chinese Medicine

基　　金：国家中医临床研究基地(江苏省中医院)开放课题(JD2022SZ05);江苏省中医院高血压病临床医学创新中心(k2021j17,k2021j17-1);江苏高校优势学科建设工程资助项目(PAPD);江苏省研究生科研与实践创新计划(SJCX23_0754)。

摘　　要：目的 探究平肝益肾方治疗自发性高血压大鼠靶器官损害的效应机制。方法 使用超高效液相色谱-四极杆飞行时间质谱(UPLC-Q-TOF/MS)测定平肝益肾方的化学成分,结合网络药理学方法对平肝益肾方的化学成分进行靶标分析、功能富集,预测平肝益肾方治疗高血压及其靶器官损害的潜在作用机制。将自发性高血压大鼠随机分为模型组、平肝益肾方低剂量组(2 g·kg^(-1))、平肝益肾方高剂量组(5 g·kg^(-1))和缬沙坦组(7.2 mg·kg^-1)),每组6只。Wistar-Kyoto大鼠作为对照组,对照组和模型组大鼠灌胃生理盐水,连续灌胃8周。采用HE和Masson染色,观察大鼠心脏和胸主动脉的病理学损伤和纤维化程度。采用免疫组织化学染色和Western blot检测大鼠心脏、肝脏和肾脏中EGFR的表达水平。采用免疫荧光染色检测大鼠心脏、肝脏和肾脏中EGFR和EEA1共定位情况。结果 通过UPLC-Q-TOF/MS检测出平肝益肾方中的26种化学成分。网络药理学揭示EGFR、PIK3R1和EP300等可能是平肝益肾方治疗高血压及其靶器官损害的关键治疗靶点,平肝益肾方治疗高血压及其靶器官损害可能与EGFR酪氨酸激酶抑制剂耐药性、血脂与动脉粥样硬化和HIF-1信号通路等密切相关。平肝益肾方高剂量组可显著降低大鼠的收缩压和平均动脉压(P<0.05,P<0.01),减轻心脏和胸主动脉的病理学损伤和纤维化程度(P<0.01,P<0.001),显著降低大鼠肝脏和肾脏中EGFR的表达水平(P<0.01),改善肝脏与肾脏的纤维化,显著降低肾脏中EGFR和EEA1的共定位水平(P<0.001),调节EGFR在早期内体的堆积,改善肾脏的纤维化。结论 整合UPLC-Q-TOF/MS、网络药理学和自发性高血压大鼠模型初步探索平肝益肾方治疗高血压及其靶器官损害的效应机制,为平肝益肾方治疗高血压进一步的机制研究和临床应用提供科学依据。OBJECTIVE To investigate the mechanisms by which Pinggan Yishen Decoction(PGYSD)contributes to alleviating target organ damage in spontaneously hypertensive rats.METHODS The chemical components of PGYSD were determined by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF/MS)and were analyzed by target analysis and functional enrichment combined with network pharmacology methods to predict the potential mechanism of PGYSD in treating hypertension and its target organ damage.Spontaneously hypertensive rats were randomly divided into the model group,low-dose PGYSD group(2 g·kg^(-1)),high-dose PGYSD group(5 g·kg^(-1)),and valsartan group(7.2 mg·kg^(-1)),with 6 rats in each group.Wistar-Kyoto rats were used as the control group,and the control group and the model group were gavaged with normal saline for 8 consecutive weeks.HE and Masson staining were used to observe the pathological damage and fibrosis degree of rat heart and thoracic aorta.Immunohistochemical staining and Western blot were used to detect the expression level of EGFR in the heart,liver and kidney of rats.Immunofluorescence staining was used to detect the co-localization of EGFR and EEA1 in the heart,liver and kidney of rats.RESULTS Twenty-six components of PGYSD were detected by UPLC-Q-TOF/MS.Network pharmacology revealed that EGFR,PIK3R1 and EP300 may be key therapeutic targets of action of PGYSD for the treatment of hypertension and its target organ damage,and that the treatment of hypertension and its target organ damage by PGYSD may be closely related to EGFR tyrosine kinase inhibitor resistance,lipids and atherosclerosis and HIF-1 signaling pathway.The high-dose group of PGYSD significantly reduced systolic blood pressure and mean blood pressure in rats(P<0.05,P<0.01),attenuated pathological damage and fibrosis in the heart and thoracic aorta(P<0.01,P<0.001),significantly reduced the expression level of EGFR in the liver and kidney of rats(P<0.01),and treated fibrosis in liver and kidney,reduced th

关 键 词：平肝益肾方 高血压 靶器官损害 UPLC-Q-TOF/MS 网络药理学 

分 类 号：R285.5[医药卫生—中药学]