基于肝脏时钟基因bmal1、clock和per2探讨清达颗粒降压的作用机制  

Antihypertensive Mechanism of Qingda Granule Based on Liver Clock Genes bmal1,clock and per2

作　　者：褚剑锋[1,2] 王天一 鲁竹青 王乙连 卢妍 彭美中 彭军 陈可冀[3] CHU Jianfeng;WANG Tianyi;LU Zhuqing;WANG Yilian;LU Yan;PENG Meizhong;PENG Jun;CHEN Keji(Fujian University of Traditional Chinese Medicine,Fuzhou 350100,Fujian,China,Fujian Key Laboratory of Integrative Medicine on Geriatrics,Fuzhou 350100,Fujian,China)

机构地区：[1]福建中医药大学,福州350100 [2]福建省中西医结合老年性疾病重点实验室,福州350100 [3]中国中医科学院西苑医院

出　　处：《中西医结合心脑血管病杂志》2021年第24期4266-4275,共10页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：国家自然科学基金项目(No.81774135);福建省自然科学基金项目(No.2018J01884);福建省卫计委医学创新人才项目(No.2018-CX-42)。

摘　　要：目的观察清达颗粒对自发性高血压大鼠(SHR)血压的影响,并分析其对肝脏bmal1、clock和per2蛋白及mRNA表达的影响。方法将16只4周龄SHR随机分为模型组(SHR组)和清达颗粒组(SHR+QDG组),同时将8只同周龄Wistar Kyoto大鼠作为空白组(WKY组)。SHR+QDG组给予清达颗粒0.9 g/(kg·d),每次1 mL,连续给药10周,SHR组及WKY组每日给予等量生理盐水灌胃。每周测量1次血压,用药结束时采用M型超声检测大鼠心脏和腹主动脉结构和功能,计算左室射血分数(LVEF)、左室短轴缩短率(LVFS)和腹主动脉脉冲波传导速度;苏木精-伊红和Masson染色法观察3组大鼠腹主动脉及肝脏组织病理学改变;转录组测序及基因本体(GO)、京都基因与基因组百科全书(KEGG)富集分析清达颗粒对SHR肝脏干预的信号通路。实时荧光定量聚合酶链式反应(RT-qPCR)检测3组大鼠肝脏组织bmal1、clock和per2 mRNA表达;免疫组织化学法检测肝脏bmal1、clock和per2蛋白表达。结果清达颗粒能明显抑制SHR收缩压、舒张压升高趋势,降低LVEF和LVFS(P<0.05),抑制大鼠腹主动脉脉冲波传导速度增高(P<0.05),减轻腹主动脉内膜结构、厚度、纤维化程度,抑制肝脏昼夜节律基因per2表达。结论清达颗粒可明显降低SHR血压,改善心脏收缩功能,减轻动脉血管重构,抑制肝脏昼夜节律基因per2表达。Objective To observe the effect of Qingda Granule on blood pressure of spontaneously hypertensive rats(SHR)and its effects on the expressions of bmal1,clock,and per2 protein and mRNA in liver.Methods Sixteen 4-week-old SHR were randomly divided into model group(SHR group),Qingda Granule group(SHR+QDG group),and 8 Wistar Kyoto rats with the same age were selected as blank group(WKY group).SHR+QDG group was gavaged with Qingda Granule 0.9g/(kg·d)for 10 weeks,SHR group and WKY group were gavaged with the same volume of saline daily.Blood pressure was measured once a week.At the end of medication,the structure and function of heart,and abdominal aorta were detected by M-mode ultrasound,left ventricular ejection fraction(LVEF),left ventricular fraction shortening(LVFS),and pulse wave conduction velocity of abdominal aorta were tested.The histopathological changes of abdominal aorta and liver were observed by hematoxylin-eosin and Masson staining.The signaling pathways of Qingda Granules for SHR liver were analyzed by transcriptome sequencing,Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment.The mRNA expressions of bmal1,clock and per2 were detected by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR),and the protein expressions of bmal1,clock and per2 were detected by immunohistochemistry.Results Qingda Granule could significantly inhibit the increasing trend of SHR systolic and diastolic blood pressure(P<0.05),and decrease of LVEF and LVFS(P<0.05).It could inhibit the increase of pulse wave conduction velocity of abdominal aorta in rats(P<0.05).Qingda Granule could relieve the intima,thickness and fibrosis degree of abdominal aorta,and inhibit the expression of liver circadian gene per2.Conclusion Qingda Granule could significantly reduce the blood pressure of SHR,improve cardiac systolic function,relieve arterial remodeling,and inhibit the expression of liver circadian gene per2.

关 键 词：自发性高血压 清达颗粒 血压 心脏功能 血管病理形态 昼夜节律 大鼠 实验研究 

分 类 号：R54[医药卫生—心血管疾病]