橙皮素对大鼠肾内动脉肌原性反应的影响及其机制  

Effects and mechanisms of hesperetin on myogenic response of rat intrarenal artery

作　　者：张蕾[1] 邢国芳 郭鹏美 刘宇[1] ZHANG Lei;XING Guo-fang;GUO Peng-mei;LIU Yu(Department of Pharmacology,Shanxi Medical University,Taiyuan 030001,Shanxi Province,China)

机构地区：[1]山西医科大学药理学教研室,山西太原030001

出　　处：《中国临床药理学杂志》2021年第5期540-543,共4页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(81603111);山西省“1331工程”重点学科建设计划基金资助项目[晋教科(2017)6号]。

摘　　要：目的探讨橙皮素(HSP)对离体大鼠肾内动脉(RIA)肌原性反应的影响,并分析其与蛋白激酶C(PKC)、内向整流钾通道(Kir)的相关性。方法将雄性SD大鼠断头处死后,取其肾脏放于生理盐溶液(PSS)中,显微镜下分离并剪取2 mm的RIA血管环用于微血管张力测定实验。血管环抑制收缩实验:将同一根RIA的不同段随机分为KCl收缩组、PE收缩组、VP收缩组、BaCl2收缩组(加入30μmol·L^(-1)HSP预孵30 min后分别加入60 mmol·L^(-1)KCl,10μmol·L^(-1)PE,3μmol·L^(-1)VP,0.3 mmol·L^(-1)BaCl24种收缩剂直至RIA达到收缩坪台);舒张机制实验:将同一根RIA的不同段随机分为KCl预收缩组、PE预收缩组、VP预收缩组、BaCl2预收缩组(预收缩组中加入上述相同浓度的4种收缩剂收缩RIA后,加入PKA、PKC抑制剂各1μmol·L-1预孵15 min,30μmol·L^(-1)HSP舒张RIA)。膜片钳实验:将RIA平滑肌细胞(SMCs)Kir电流密度分为BaCl2联合组、GO6983联合组;实时荧光定量聚合酶链反应(qRT-PCR):将RIA分为对照组和HSP组(预孵30μmol·L^(-1)HSP 8h)。用血管环实验记录RIA血管张力的变化,用膜片钳实验记录RIA SMCs Kir电流密度的变化,用qRT-PCR检测Kir2.1和PKC-α、PKC-β基因表达水平。结果KCl收缩组,PE收缩组,VP收缩组,BaCl2收缩组RIA收缩百分率分别为(24.25±6.11)%,(20.09±12.67)%,(48.14±1.68)%,(15.26±6.66)%;KCl预收缩组,PE预收缩组,VP预收缩组,BaCl2预收缩组的舒张百分率分别为(21.35±5.55)%,(27.84±8.64)%,(23.30±12.01)%,(19.22±2.22)%;BaCl2联合组、GO6983联合组RIA SMCs Kir电流分别为(-4.29±0.26),(-10.65±1.10)pA/pF;Kir 2.1通道、PKC-α、PKC-β的mRNA相对表达量分别为1.91±0.23,0.52±0.29,0.69±0.14。组间比较,差异均有统计学差异(均P<0.05)。结论HSP有抑制RIA收缩的作用,且HSP该作用与舒张预收缩RIA的作用均可能与Kir通道的开放和PKC介导的通路有关。Objective To investigate the effect of hesperetin(HSP)on myogenesis of the rat internal renal artery(RIA),and to analyze its relationship to protein kinase C(PKC)/Inward rectifier K+channels(Kir)signaling pathway.Methods All SD male rats were euthanized by beheaded executed,then the kidneys were removed and transferred in physiological salt solution(PSS).The RIAs were separated under the microscope and cut into 2 mm vascular rings for microvascular tension measurement.Inhibition of diastolic test:the different segments of a RIAwere randomly divided into KCl-contraction group,PE-contraction group,VP-contraction group,BaCl2-contraction group(60 mmol·L-1KCl,10μmol·L^(-1)PE,3μmol·L^(-1)VP,0.3 mmol·L-1BaCl2 were added into chamber respectively after preincubation of 30μmol·L^(-1)HSP for 30 min);relaxation mechanism text:a RIA was divided into KCl-precontracted group,PE-precontracted group,VP-precontracted group,BaCl2-precontracted group(60 mmol·L^(-1)KCl,10μmol·L^(-1)PE,3μmol·L^(-1)VP,0.3mmol·L^(-1)BaCl2 were added into chamber respectively,after reaching plateau,30μmol·L^(-1)HSP was added into chamber after 15 min incubation with 1μmol·L^(-1)PKC and PKA inhibitor);patch clamp experiment:Kir current density were divided into BaCl2 combined group,GO6983 combined group.Real-time quantitative polymerase chain reaction:total RIAs were divided into control group and HSP group(preincubation with 30μmol·L^(-1)HSP for 8 h).The reaction of vascular tension was detected by vascular ring experiment.The change of Kir current density was detected by Patch clamp experiment,and the expression levels of Kir2.1,PKC-α,PKC-βwere detected by qRT-PCR.Results The contraction percentage of KCl-contraction group,PE-contraction group,VP-contraction group,BaCl2-contraction group were(24.25±6.11)%,(20.09±12.67)%,(48.14±1.68)%,(15.26±6.66)%;the relaxation percentage of KCl-precontracted group,PE-precontracted group,VP-precontracted group,BaCl2-precontracted group were(21.35±5.55)%,(27.84±8.64)%,(23.30±12.01)%,(19.22�

关 键 词：橙皮素 肾内动脉 抑制收缩 Kir通道 蛋白激酶C 

分 类 号：R28[医药卫生—中药学]