肾周白色脂肪组织源外泌体远距调控大鼠血压  

Perirenal adipose tissue derived exosomes regulate blood pressure in rats

作　　者：张培森 梁沥文 施真[1] ZHANG Peisen;LIANG Liwen;SHI Zhen(Department of Physiology of Binzhou Medical University,Yantai 264003,China;Weihai Central Blood Station,Weihai 264200)

机构地区：[1]滨州医学院生理学教研室,山东烟台264003 [2]威海市中心血站,山东威海264200

出　　处：《中国比较医学杂志》2023年第3期51-58,共8页Chinese Journal of Comparative Medicine

基　　金：山东省自然科学基金(ZR2018MC011)。

摘　　要：目的研究肾周白色脂肪组织作为内分泌器官是否以外泌体囊泡作为运载体调控大鼠动脉血压。方法采用成年自发性高血压大鼠(spontaneously hypertensive rat,SHR)和正常血压的Wistar-Kyoto(WKY)大鼠。将WKY大鼠和SHR的肾周白色脂肪置于无血清(无外泌体)培养基中培养,使用外泌体分离提取试剂盒(exoEasy Maxi Kit)分离分泌囊泡。经纳米粒径测定、透射电镜和表面标志物鉴定符合外泌体囊泡(Exosomes,Exo)特征后用于后续实验。SHR或WKY大鼠各随机分为两组,分别尾静脉注射SHR大鼠外泌体(Exo-SHR)或WKY大鼠外泌体(Exo-WKY),每周1次,连续6周。在清醒状态下使用无创尾动脉加压系统测量动脉血压,每周1次。注射后第6周末进行急性实验。采用颈动脉插管法进行动脉血压直接测定,采用HE染色和Masson染色观察心血管病理重构。外周交感神经张力(peripheral sympathetic tone,PST)高低以静脉注射盐酸六烃季铵引起的降压反应与Exo-WKY处理的WKY大鼠的降压反应的比值大小表示。另外,PKH67绿色荧光染料标记外泌体后尾静脉注射进行体内示踪,与原代培养的神经元孵育进行体外示踪。结果肾周白色脂肪组织分泌的外泌体囊泡经透射电镜观察到典型外泌体半托杯状囊泡结构,纳米粒径检测平均直径集中在100 nm左右,且具有表面特异标志物肿瘤易感基因101(tumor susceptibility gene,TSG101)和白细胞分化9(cluster of differentiation 9,CD9)表达。SHR组收缩压(Systolic blood pressure,SBP)在注射Exo-WKY后的第2周开始下降,并在整个干预过程中保持低于注射前的水平(P<0.05)。应用Exo-WKY可减少SHR心肌细胞横截面积、主动脉中膜厚度及降低PST(P<0.05)。而WKY大鼠应用Exo-SHR后,SBP、心肌细胞横截面积、主动脉中膜厚度及PST均未见明显变化(P>0.05)。在体示踪发现PKH67标记外泌体囊泡经尾静脉注射24 h后,在下丘脑室旁核(paraventricular nucleus,PVH)、孤束�ObjectiveTo investigate whether perirenal adipose tissues as an endocrine organ regulate arterial blood pressure in rats via exosomes.MethodsSpontaneously hypertensive rats(SHR)and normotensive Wistar-Kyoto(WKY)rats were used in this study.Perirenal fat of normotensive WKY rats and SHRs was cultured in serum-free(exosome-free)medium,and secretory vesicles were isolated using an exoEasy Maxi Kit.The isolated vesicles were identified by nano-size measurement,transmission electron microscopy,and surface markers,which were consistent with the characteristics of exosomes(Exo)and used for subsequent experiments.SHR and WKY rats were randomly divided into two groups and injected with Exo-SHR or Exo-WKY via the tail vein once a week for 6 weeks.Arterial blood pressure was measured while rats were conscious using a non-invasive caudal cuff system.Acute experiments were conducted at the end of week 6 after the injection of exosome-like vesicles.Carotid artery cannulation was used to directly measure arterial blood pressure.HE and Masson staining was used to observe cardiovascular pathological remodeling.Peripheral sympathetic tone(PST)was expressed as the relative ratio of the depression response induced by intravenous injection of hexamethonium hydrochloride to that in WKY rats treated with Exo-WKY.Exosomes labeled with PKH67 green fluorescent dye were injected into the tail vein for in vivo tracing,and then incubated with primary cultured neurons for in vitro tracing.ResultsExosomes secreted from perirenal white adipose tissue were typical exosome semi-cup-like vesicles under transmission electron microscopy.The average diameter of nanoparticles was approximately 100 nm,and surface-specific markers TSG101 and CD9 were expressed.The systolic blood pressure(SBP)of SHRs began to decrease in the second week after injection of Exo-WKY and remained low throughout experiment(P<0.05).Exo-WKY also decreased the cross-sectional area of cardiomyocytes,aortic media thickening,and PST in SHRs(P<0.05).However,after administering Exo-S

关 键 词：自发性高血压 肾周白色脂肪 外泌体 交感神经活动 

分 类 号：R-33[医药卫生]