黄葵胶囊预处理人脐带间充质干细胞外泌体改善肾脏缺血-再灌注损伤的作用及机制  

The effect and mechanism of Huangkui capsule pretreatment of human umbilical cord mesenchymal stem cell-derived exosomes on improving renal ischemia-reperfusion injury

作　　者：姚雅韡 贺佳辉 王浩[2] 王誉潼 王睿妍 万幸雨 刘雨佳 吕兴华[2] Yao Yawei;He Jiahui;Wang Hao;Wang Yutong;Wang Ruiyan;Wan Xingyu;Liu Yujia;LüXinghua(The First Clinical Medical College of Lanzhou University,Lanzhou 730000,China;不详)

机构地区：[1]兰州大学第一临床医学院,兰州730000 [2]兰州大学第一医院日间手术中心

出　　处：《器官移植》2025年第2期237-245,共9页Organ Transplantation

基　　金：甘肃省自然科学基金(23JRRA0932)。

摘　　要：目的 探讨黄葵胶囊预处理的人脐带间充质干细胞(HUC-MSC)来源外泌体(Exo)对肾脏缺血-再灌注损伤(IRI)的影响及机制。方法 将HUC-MSC分别置于含有不同浓度的黄葵胶囊培养基中培养24 h,测定细胞活力,选取适宜浓度进行后续实验。选取50μg/mL的黄葵胶囊预处理HUC-MSC 24 h,使用Exo提取试剂盒提取Exo,透射电镜观察其形态、纳米粒径分析检测粒径大小、蛋白质印迹法检测Exo膜表面标记蛋白表达。将人肾小管上皮细胞(HK-2细胞)随机分为缺氧/复氧组(M组)、缺氧/复氧+Exo组(E组)和缺氧/复氧+黄葵胶囊预处理Exo组(H组)。蛋白质印迹法测定内质网应激(ERS)相关蛋白表达,实时荧光定量聚合酶链反应测定ERS相关基因信使RNA(mRNA)表达。将小鼠随机分为假手术组(Sham组)、缺血-再灌注组(I/R组)、缺血-再灌注+Exo组(E组)和缺血-再灌注+黄葵胶囊预处理Exo组(H组)。24 h后行肾脏组织学评估,血清肌酐(Scr)、血尿素氮(BUN)测定及炎症因子检测。结果 Exo和黄葵胶囊预处理Exo均具有双层膜结构,呈杯状形态;二者平均粒径大小分别为116.8 nm和81.3 nm;两者均表达CD9、CD63、TSG101。与M组比较,E组转录激活因子6(ATF6)、蛋白激酶R样内质网激酶(PERK)蛋白相对表达量下降,m RNA相对表达量升高,C/EBP同源蛋白(CHOP)蛋白相对表达量升高,mRNA相对表达量下降;与E组相比,H组ATF6、PERK、CHOP蛋白相对表达量均降低,ATF6、PERK mRNA相对表达量下降(均为P<0.05)。动物实验结果显示,与Sham组相比,I/R组肾小管损伤评分升高,Scr、BUN、白细胞介素(IL)-1β、IL-10、IL-18、肿瘤坏死因子(TNF)-α水平升高;与I/R组比较,E组和H组肾小管损伤评分降低,Scr、BUN、IL-1β、IL-10、IL-18、TNF-α水平下降;与E组比较,H组肾小管损伤评分下降,Scr、BUN、IL-1β、IL-10、IL-18、TNF-α水平下降(均为P<0.05)。结论 黄葵胶囊预处理HUC-MSC来源的Exo可通过抑制ERS,改善肾脏IRI。Objective To explore the effects and mechanisms of human umbilical cord mesenchymal stem cell(HUC-MSC)-derived exosomes(Exo)pretreated with Huangkui capsules on renal ischemia-reperfusion injury(IRI).Methods HUC-MSCs were cultured in media containing different concentrations of Huangkui capsules for 24 hours to determine cell viability and select an appropriate concentration for subsequent experiments.HUC-MSCs were pretreated with 50μg/mL Huangkui capsules for 24 hours,and Exo were extracted using an exosome extraction kit.The morphology was observed under a transmission electron microscope,particle size was measured by nanoparticle tracking analysis,and the expression of exosomal membrane surface marker proteins was detected by Western blot.Human renal tubular epithelial cells(HK-2 cells)were randomly divided into hypoxia/reoxygenation group(M group),hypoxia/reoxygenation+Exo group(E group),and hypoxia/reoxygenation+Huangkui capsules pretreated Exo group(H group).Western blotting was used to measure the expression of endoplasmic reticulum stress(ERS)-related proteins,and real-time fluorescent quantitative reverse transcription polymerase chain reaction was used to measure the expression of ERS-related gene messenger RNA(mRNA).Mice were randomly divided into sham operation group(Sham group),ischemia-reperfusion group(I/R group),ischemia-reperfusion+Exo group(E group),and ischemia-reperfusion+Huangkui capsules pretreated Exo group(H group).Renal histological assessment,serum creatinine(Scr),blood urea nitrogen(BUN)measurement and inflammatory factor detection were performed 24 hours later.Results Both Exo and Huangkui capsules prereated Exo had a bilayer membrane structure and a cup-shaped morphology;their average particle sizes were 116.8 nm and 81.3 nm,respectively.Both expressed CD9,CD63,TSG101.Compared with the M group,the E group had decreased relative expression of transcription factor 6(ATF6)and protein kinase R-like endoplasmic reticulum kinase(PERK)proteins,increased mRNA relative expression,increased re

关 键 词：肾移植 缺血-再灌注损伤 黄葵胶囊 人脐带间充质干细胞 外泌体 内质网应激 急性肾损伤 炎症因子 

分 类 号：R617[医药卫生—外科学] R28[医药卫生—临床医学]