摘要
目的研究骨髓间充质干细胞外泌体(BMSCs-exo)对高血压大鼠血管内皮的影响及机制。方法体外分离培养获取大鼠骨髓间充质干细胞(BMSCs),诱导形成BMSCs-exo。选取12周龄雄性自发性高血压大鼠(SHR)40只,随机分为BMSCs-exo组(10只)、对照组(10只)、miR-132-3p高表达组(10只)、miR-132-3p空载组(10只),另选取雄性WKY大鼠为健康组(10只)。BMSCs-exo组予以尾静脉注射BMSCs-exo,每次30μg,隔日1次,连续注射7次;miR-132-3p高表达组与miR-132-3p空载组分别以miR-132-3p高表达慢病毒颗粒和空载体慢病毒颗粒尾静脉注射,每次0.1 mL,3 d注射1次,至BMSCS-exo组干预结束;健康组、对照组以同法注射生理盐水。末次干预后采用实时荧光定量聚合酶链式反应(qRT-PCR)检测各组miR-132-5p表达变化。测量各组大鼠尾动脉收缩压,检测血清内皮素-1(ET-1)、一氧化氮(NO)、血栓素B_(2)(TXB_(2))、前列环素(PGI_(2))水平。结果倒置显微镜下观察,BMSCs形态从圆形逐渐向梭形或多角形生长,流式细胞术鉴定BMSCs表面抗原符合生物学特征。透射电镜及蛋白印迹法验证提取的囊泡状物质为外泌体。与对照组比较,BMSCs-exo组、miR-132-3p高表达组miR-132-3p相对表达量升高(P<0.05);与BMSCs-exo组比较,miR-132-3p高表达组miR-132-3p相对表达量升高(P<0.05),miR-132-3p空载组相对表达量降低(P<0.05);与对照组比较,BMSCs-exo组、miR-132-3p高表达组尾动脉收缩压、血浆ET-1及TXB_(2)水平降低,血浆NO、PGI_(2)水平升高(P<0.05);与BMSCs-exo组和miR-132-3p高表达组比较,miR-132-3p空载组尾动脉收缩压、血浆ET-1及TXB_(2)水平升高,血浆NO、PGI_(2)水平降低(P<0.05)。对照组大鼠胸主动脉血管壁增厚、管腔狭窄,动脉壁中层平滑肌细胞肥大,细胞结构排列紊乱;BMSCs-exo组、miR-132-3p高表达组病理变化较对照组轻,但BMSCs-exo组与miR-132-3p高表达组病理变化相当。结论BMSCs-exo能降低高血压大鼠血压,改善血管内皮功能,抑制血管病理变化,其机制可能与提高miR-132-3p的表达有关。
Objective To study the effect and mechanism of bone marrow mesenchymal stem cell exosomes(BMSCs-exo)on the vascular endothelium of hypertensive rats.Methods Rat bone marrow mesenchymal stem cells(BMSCs)were isolated and cultured in vitro to induce the formation of BMSCs-exo.Forty 12-week-old male SHR rats were selected and randomly divided into BMSCs-exo group(10 rats),control group(10 rats),miR-132-3p high expression group(10 rats),miR-132-3p empty group(10 rats),and male WKY rats as healthy group(10 rats).The rats in BMSCs-exo group were injected with BMSCs-exo via tail vein,30μg/time,once every other day,7 times continuously.The rats in miR-132-3p high expression group and miR-132-3p empty group were injected with the miR-132-3p high expression lentiviral particles and the empty vector chronic disease particles via tail vein,respectively,0.1 mL/time,once every 3 days.The rats in healthy group and control group were injected with saline in the same way.After the last intervention,the expression of miR-132-5p was detected by quantitative real-time PCR(qRT-PCR).Systolic blood pressure(SBP)of the tail artery of each group was measured,and the levels of plasma endothelin-1(ET-1),nitric oxide(NO),thromboxane B_(2)(TXB_(2)),and prostacyclin(PGI_(2))were detected.Results Under an inverted microscope,the BMSCs gradually grew from round to fusiform or polygonal.The surface antigen of BMSCs was identified by flow cytometry,which was consistent with the biological characteristics.Transmission electron microscopy and Western Blot confirmed that the vesicular substance was exo.Compared with the control group,the relative expression of miR-132-3p was higher in BMSCs-exo group and miR-132-3p high expression group(P<0.05).Compared with the BMSCs-exo group,the relative expression level of miR-132-3p in miR-132-3p high expression group was higher,and that in miR-132-3p empty group was lower(P<0.05).Compared with miR-132-3p high expression group,the relative expression of miR-132-3p in miR-132-3p empty group was lower(P<0.05).Compared with control group,the tail artery SBP,plasma ET-1,and TXB_(2) levels in BMSCs-exo group and miR-132-3p high expression group were lower,and the plasma NO and PGI_(2) levels were higher(P<0.05).Compared with BMSCs-exo group and miR-132-3p high expression group,the tail artery SBP,plasma ET-1,and TXB_(2) levels in miR-132-3p empty group were higher,and the plasma NO and PGI_(2) levels were lower(P<0.05).In control group,the thoracic aorta was thickened,the lumen was narrowed,the smooth muscle cells in the middle layer of the artery wall were hypertrophy,and the cell structure was disordered.The pathological changes of BMSCs-exo group and miR-132-3p high expression group were lighter than those of control group,but the pathological changes of the two groups were similar.Conclusion BMSCs-exo can reduce blood pressure,improve vascular endothelial function,and inhibit vascular pathological changes in hypertensive rats.The mechanism may be related to the increase of miR-132-3p expression.
作者
郑茜
张勇
杨东伟
ZHENG Xi;ZHANG Yong;YANG Dongwei(Zhengzhou Central Hospital Affiliated to Zhengzhou University,Zhengzhou 450007,Henan,China)
出处
《中西医结合心脑血管病杂志》
2021年第22期3891-3896,共6页
Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease
基金
河南省医学科技攻关计划(联合共建)项目(No.LHGJ20191058)。
关键词
高血压
骨髓间充质干细胞
外泌体
血管内皮功能
自发性高血压大鼠
实验研究
hypertension
bone marrow mesenchymal stem cells
exosomes
vascular endothelial function
spontaneously hypertensive rats
experiment research
作者简介
郑茜,E-mail:dsjt2345@sina.com。
