摘要
目的 探索脂肪干细胞(ADSCs)来源外泌体(Exo)负载mi RNA-21调控糖尿病肾病(DN)大鼠肾脏内皮-间充质转化(End MT)的机制。方法 选择3月龄SPF级SD雄性大鼠70只,其中10只用于提取ADSCs,60只用于实验研究。选择对数生长期的ADSCs,并与mi RNA-21过表达、低表达及空载体质粒进行脂质体转染后提取Exo,同时提取不转染任何质粒ADSCs的Exo。60只大鼠按照随机抓取法分为空白对照组、模型组、ADSCs-Exo组、ADSCs-Exo-NC组、ADSCs-Exo-过表达组、ADSCs-Exo-低表达组,每组10只。ADSCs-Exo组、ADSCs-Exo-NC组、ADSCs-Exo-过表达组及ADSCs-Exo-低表达组分别在复制模型前1周于尾静脉注射相应Exo,剂量为10 mg/kg,1次/d,连续1周。除空白对照组外,其余各组大鼠进行高糖高脂饮食和链脲佐菌素溶液腹腔注射处理建立DN模型。观察各组大鼠肾小球病理情况;比较各组大鼠内皮细胞凋亡率,尿微量白蛋白、血肌酐水平;比较各组大鼠E-钙黏附蛋白(E-cadherin)、α-平滑肌肌动蛋白(α-SMA)、铁死亡抑制蛋白1(FSP1)蛋白表达。结果 空白对照组肾小球结构完整;ADSCs-Exo组、ADSCs-Exo-NC组与模型组肾小球内皮细胞结构紊乱;ADSCs-Exo-过表达组可见少量内皮细胞;ADSCs-Exo-低表达组出现新生内皮细胞。与造模后比较,造模后4周模型组、ADSCs-Exo组、ADSCs-Exo-NC组、ADSCs-Exo-过表达组尿微量白蛋白、血肌酐水平升高,ADSCs-Exo-低表达组尿微量白蛋白、血肌酐水平降低(P<0.05)。与空白对照组比较,模型组造模后和造模后4周尿微量白蛋白、血肌酐水平升高(P<0.05)。与ADSCs-Exo-NC组比较,ADSCs-Exo-过表达组造模后4周尿微量白蛋白和血肌酐升高,ADSCs-Exo-低表达组尿微量白蛋白、血肌酐水平降低(P<0.05)。与空白对照组比较,模型组内皮细胞凋亡率升高(P<0.05);与ADSCs-Exo-NC组比较,ADSCs-Exo-过表达组内皮细胞凋亡率升高,ADSCs-Exo-低表达组内皮细胞凋亡率降低(P<0.05)。与空白对照组比较,模型组α-SMA、FSP1蛋白表达量升高,E-cadherin蛋白表达量降低(P<0.05)。与ADSCs-Exo-NC组比较,ADSCs-Exo-过表达组α-SMA、FSP1蛋白表达量升高,E-cadherin蛋白表达量降低(P<0.05);ADSCs-Exo-低表达组α-SMA、FSP1蛋白表达量降低,E-cadherin蛋白表达量升高(P<0.05)。结论 ADSCs来源Exo负载mi RNA-21可以促进DN大鼠肾小球End MT的发生。
Objective To investigate the mechanism of adipose stem cells(ADSCs)derived exosomes(Exo)-loaded miRNA-21 in regulating renal endothelial-to-mesenchymal transition(EndMT)of rats with diabetic nephropathy(DN).Methods Seventy 3-month-old SPF-grade male SD rats were selected,among which 10 were used for extracting ADSCs and 60 were used for experimental research.ADSCs in the logarithmic growth phase were selected,and Exo were extracted after liposome transfection with miRNA-21 overexpression,low expression,and empty plasmid;meanwhile,Exo of ADSCs without transfection of any plasmid was extracted.Sixty rats were divided into blank control group,model group,ADSCs-Exo group,ADSCs-Exo-NC group,ADSCs-Exo-overexpression group,and ADSCs-Exo-low expression group according to the random grasping method,with 10 rats in each group.The ADSCs-Exo group,ADSCs-Exo-NC group,meanuhile,ADSCs-Exo-overexpression group,and ADSCs-Exo-low expression group were respectively injected with the corresponding Exo through the tail vein one week before replicating the model at a dose of 10 mg/kg once a day for one consecutive week.Except for blank control group,the other groups of rats were treated with a high-sugar and high-fat diet and intraperitoneal injection of Streptozotocin Solution to establish the DN model.The pathological conditions of glomerulus of rats in each group were observed;the apoptosis rate of endothelial cells,the levels of urine microalbumin and serum creatinine of rats in each group were compared;and the protein expressions of E-cadherin,α-smooth muscle actin(α-SMA),and ferroptosis inhibitory protein 1(FSP1)of rats in each group were compared.Results The glomerular structure of blank control group was intact;while the structures of glomerular endothelial cells in ADSCs-Exo group,ADSCs-Exo-NC group,and model group were disordered;a small number of endothelial cells were observed in ADSCs-Exo-overexpression group;while new endothelial cells appeared in ADSCs-Exo-low expression group.Compared with after modeling,the levels of urinary microalbumin and serum creatinine increased in model group,ADSCs-Exo group,ADSCs-Exo-NC group,and ADSCS-Exo-overexpression group 4 weeks after modeling,while those in ADSCs-Exo-low expression group decreased(P<0.05).Compared with blank control group,the levels of urinary microalbumin and serum creatinine in model group increased after modeling and 4 weeks after modeling(P<0.05);compared with ADSCs-Exo-NC group,the levels of urinary microalbumin and serum creatinine increased in ADSCs-Exo-overexpression group,while those in ADSCS-Exo-low expression group decreased 4 weeks after modeling(P<0.05).Compared with blank control group,the apoptosis rate of endothelial cells in model group increased(P<0.05);compared with ADSCs-Exo-NC group,the apoptosis rate of endothelial cells in ADSCs-Exo-overexpression group increased,while that in ADSCs-Exo-low expression group decreased(P<0.05).Compared with blank control group,the expression levels ofα-SMA and FSP1 proteins in model group increased,while the expression level of E-cadherin protein decreased(P<0.05).Compared with ADSCs-Exo-NC group,the expression levels ofα-SMA and FSP1 proteins in ADSCs-Exo-overexpression group increased,while the expression level of E-cadherin protein decreased(P<0.05);in ADSCs-Exo-low expression group,the expression levels ofα-SMA and FSP1 proteins decreased,while the expression level of E-cadherin protein increased(P<0.05).Conclusion ADSCs derived Exo-loaded miRNA-21 can promote the occurrence of EndMT in glomeruli of DN rat.
作者
热依汗·阿不都力提甫
张琳
古丽鲜·吐尔洪
李光玉
Reyihan Abudulitifu;ZHANG Lin;Gulixian Tuerhong;LI Guangyu(Department of Nephrology,the Second Affiliated Hospital of Xinjiang Medical University,Xinjiang Uygur Autonomous Region,Urumqi 830028,China)
出处
《中国医药导报》
2025年第31期13-18,共6页
China Medical Herald
基金
新疆少数民族科技人才特殊培养计划科研项目(2021D03023)。
