钩藤降压解郁方对高血压并发抑郁症大鼠海马突触超微结构及TLR4、NF-κB p65和NLRP3蛋白表达的影响  

Effect of Gouteng Jiangya Jieyu Formula(钩藤降压解郁方)on the Ultrastructure of Hippocampal Synapses and the Expression of TLR4,NF-κB p65 and NLRP3 Proteins of Hippocampus in Rats of Hypertension Complicated with Depression

作　　者：朱智恒 赵红霞 刘叶倩 陈蕾 黄铃格 李弘 马丹凤[4] 陈春茗 曾水清 任卫琼[1] ZHU Zhiheng;ZHAO Hongxia;LIU Yeqian;CHEN Lei;HUANG Lingge;LI Hong;MA Danfeng;CHEN Chunming;ZENG Shuiqing;REN Weiqiong(The First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha Hunan 410007,China;Chongqing Wanzhou Institute for Food and Drug Control,Chongqing 404000,China;The Second People's Hospital of Anhui Province,Hefei Anhui 230041,China;Hunan Children's Hospital,Changsha Hunan 410007,China)

机构地区：[1]湖南中医药大学第一附属医院,湖南长沙410007 [2]重庆市万州食品药品检验所,重庆万州404000 [3]安徽省第二人民医院,安徽合肥230041 [4]湖南省儿童医院,湖南长沙410007

出　　处：《中医药导报》2024年第8期40-46,共7页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：湖南省自然科学基金项目(2021JJ80065);湖南省中药粉体与创新药物研究省部共建国家重点实验室培育基地开放基金项目(21PTKF1010);湖南省中医药科研课题项目(A2023019);湖南省卫生健康委员会科研计划课题项目(C202313056551,D202313057728)。

摘　　要：目的:研究钩藤降压解郁方对高血压并发抑郁症(HD)大鼠学习记忆功能、海马突触超微结构、海马炎症反应及海马Toll样受体4(TLR4)、核因子κB p65(NF-κB p65)和NOD样受体蛋白3(NLRP3)蛋白表达的影响。方法:将35只自发性高血压大鼠随机分为模型组、阳性对照组[苯磺酸左旋氨氯地平(0.45 mg/kg)+盐酸氟西汀(1.80 mg/kg)]和钩藤降压解郁方低、中、高(6.35、12.69、25.38 g/kg)剂量组,另取7只SD大鼠作空白对照组。采用慢性不可预知应激联合孤养的方法干预自发性高血压大鼠复制HD模型。造模同时灌胃给药,1次/d,连续6周。用无创血压计测量尾动脉收缩压和舒张压;Morris水迷宫检测学习记忆能力;透射电镜观察海马突触超微结构;酶联免疫吸附法检测海马炎症因子白介素-1β(IL-1β)、IL-18的水平;免疫组化法检测海马CA1区中TLR4、NF-κB p65和NLRP3蛋白的表达。结果:给药干预结束后,与空白对照组比较,模型组大鼠尾动脉收缩压和舒张压较高(P<0.01);模型组大鼠逃避潜伏期和第一次跨越平台时间延长,目标象限总时间和目标象限总路程减少(P<0.01);模型组大鼠海马突触超微结构模糊,部分突触结构出现融合,突触内线粒体嵴肿胀断裂,突触小泡明显减少,突触后致密物质厚度较小(P<0.01);模型组大鼠海马组织IL-1β、IL-18含量及TLR4、NF-κB p65、NLRP3蛋白表达均升高(P<0.05或P<0.01)。与模型组比较,阳性对照组和钩藤降压解郁方低、中剂量组大鼠收缩压和舒张压较低(P<0.05或P<0.01);阳性对照组和钩藤降压解郁方低、中、高剂量组大鼠逃避潜伏期和第一次跨越平台时间较短,目标象限总时间和目标象限总路程增加;钩藤降压解郁方高剂量组大鼠突触结构清晰,突触前膜、后膜存在,突触前膜内突触小泡增多,突触后致密物质厚度增大(P<0.01);阳性对照组和钩藤降压解郁方中、高剂量组大鼠海马炎症因子IL-1β和IL-18Objective:To observe the effect of Gouteng Jiangya Jieyu Formula(GTJYJYF)on the learning-memory function,ultrastructure of hippocampal synapses,inflammatory response in the hippocampus as well as the expression of hippocampal TLR4,NF-κB p65 and NLRP3 proteins in spontaneously hypertensive rats complicated with depression.Methods:Totally 35 SHR rats were randomly divided into model group,positive control group[amlodipine besylate(0.45 mg/kg)+fluoxetine hydrochloride(1.80 mg/kg)],GTJYJYF low-dose group(6.35 g/kg),GTJYJYF medium-dose group(12.69 g/kg)and GTJYJYF high-dose group(25.38 g/kg),with an additional blank control group consisting of 7 Sprague-Dawley rats.The SHR rats were induced to develop hypertension complicated with depression using chronic unpredictable stress combined with isolation rearing.The modeling process involved oral administration once daily for 6 weeks.Tail arterial systolic and diastolic pressures were measured using a non-invasive blood pressure meter.Morris water maze test was conducted to evaluate learning and memory abilities.Transmission electron microscopy was used to observe the ultrastructure of hippocampal synapses.The levels of inflammatory factors(IL-1βand IL-18)in the hippocampus were measured by enzyme-linked immunosorbent assay.The expression of TLR4,NF-κB p65 and NLRP3 proteins in the CA1 region of the hippocampus was detected by immunohistochemistry.Results:After the drug intervention,compared with the blank control group,the model group exhibited higher systolic and diastolic pressures(P<0.01).The escape latency and the first time of acrossing the platform were prolonged in model group,while the total time and distance of target quadrant were decreased in model group(P<0.01).The ultrastructure of hippocampal synapses was blurred in model group,with some synaptic structures showing fusion,mitochondrial cristae swelling and rupture within synapses,a significant reduction in synaptic vesicles,and a smaller thickness of dense material after synapses(P<0.01).The levels of IL-

关 键 词：高血压并发抑郁症 钩藤降压解郁方 海马突触超微结构 Toll样受体4 核因子κB p65 NOD样受体蛋白3 学习记忆功能 大鼠 

分 类 号：R285.5[医药卫生—中药学]