人脐带间充质干细胞源小细胞外囊泡通过抑制THBS1减轻糖尿病肾病大鼠肾脏损伤  

Human umbilical cord mesenchymal stem cell-derived small extracellular vesicles attenuate kidney damage in rats with diabetic nephropathy by inhibiting THBS1

作　　者：单云洁 于洋 尹思琪 钱晖[1] SHAN Yunjie;YU Yang;YIN Siqi;QIAN Hui(School of Medicine,Jiangsu University,Zhenjiang Jiangsu 212013,China)

机构地区：[1]江苏大学医学院,江苏镇江212013

出　　处：《江苏大学学报（医学版）》2024年第6期469-475,484,共8页Journal of Jiangsu University：Medicine Edition

基　　金：国家自然科学基金面上项目(82172102);江苏省重点研发计划(社会发展)重点项目(BE2021689)。

摘　　要：目的:探究人脐带间充质干细胞来源小细胞外囊泡(hucMSCs derived small extracellular vesicles,hucMSC-sEVs)对糖尿病肾病(diabetic kidney disease,DKD)大鼠肾脏损伤的修复作用及可能机制。方法:分离培养hucMSCs并从培养上清液中提取hucMSC-sEVs。高脂联合链脲佐菌素构建DKD大鼠模型,将大鼠随机分为对照组、DKD组和hucMSC-sEVs组,每组6只,hucMSC-sEVs组在造模8周后进行尾静脉注射hucMSC-sEVs,24周后收集肾组织。肾脏组织切片进行HE染色、过碘酸雪夫(PAS)染色和天狼星红染色,观察大鼠肾组织病理学改变和纤维化样改变;蛋白质印迹检测肾组织中凋亡相关蛋白Bcl-2和Bax表达;免疫组织化学染色、免疫荧光染色和蛋白质印迹检测大鼠肾组织血小板反应蛋白1(THBS1)及其受体CD36和CD47的表达水平。结果:与对照组相比,DKD组大鼠肾脏发生明显的病理改变,如肾小球基底膜增厚和系膜增生,肾小管扩张伴有空泡变性,间质发生明显纤维化改变伴炎症细胞浸润,肾组织中Bax表达明显增加(P<0.05),Bcl-2表达显著下降(P<0.01),THBS1及其受体CD36和CD47的表达水平明显增加(P<0.001);与DKD组相比,hucMSC-sEVs组大鼠肾组织病理损伤和纤维化水平明显缓解,Bax表达显著下降(P<0.01),Bcl-2表达显著增加(P<0.01),肾脏中THBS1及其受体CD36和CD47的表达水平显著降低(P<0.001)。结论:HucMSC-sEVs可明显减轻DKD肾组织损伤发挥保护作用,其机制可能与靶向抑制THBS1的表达有关。Objective:To investigate the effect of human umbilical cord mesenchymal stem cell derived small extracellular vesicles(hucMSC-sEVs)in attenuating the damage of diabetic kidney disease(DKD)in rats.Methods:HucMSCs were isolated and cultured,and hucMSC-sEVs were extracted from the culture media supernatant.The DKD rat model was established by high fat diet combined with injection of streptozocin,rats were randomly divided into control group,DKD group,and hucMSC-sEVs group,with 6 rats in each group.The hucMSC-sEVs group received tail vein injection of hucMSC-sEVs 8 weeks after modeling,and kidney tissues were collected at week 24.Renal tissue pathology and fibrotic changes in rats were assessed using HE staining,PAS staining and Sirius Red staining.The expression of apoptosis-related proteins Bcl-2 and Bax in kidney tissues was evaluated using Western bloting.Additionally,the expression levels of THBS1 and its receptors CD36 and CD47 in rat renal tissues were determined using histochemical staining,immunofluorescence staining,and Western bloting.Results:Compared with the control group,the DKD group exhibited significant pathological changes in the kidneys,such as thickening of the glomerular basement membrane and mesangial proliferation,dilation of renal tubules accompanied by vacuolar degeneration,and marked interstitial fibrosis with infiltration of inflammatory cells.The expression of the apoptosis marker Bax in kidney tissues showed a significant increase(P<0.05),while the expression of the anti-apoptotic marker Bcl-2 exhibited a significant decrease(P<0.01).The expression levels of THBS1 and its receptors CD36 and CD47 were also significantly elevated(P<0.001).In contrast,compared with the DKD group,the hucMSC-sEVs group demonstrated marked alleviation of renal tissue pathological damage and fibrosis.The expression of Bax significantly decreased(P<0.01),while the expression of Bcl-2 significantly increased(P<0.01).Additionally,the expression levels of THBS1 and its receptors CD36 and CD47 in the kidneys were sig

关 键 词：人脐带间充质干细胞 小细胞外囊泡 糖尿病肾病 血小板反应蛋白1 

分 类 号：R587.2[医药卫生—内分泌]