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黄芪甲苷孵育的脂肪干细胞治疗糖尿病肾病 预览

Astragaloside-incubated adipose-derived stem cells for the treatment of diabetic nephropathy
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摘要 背景:研究表明,蛋白尿是糖尿病肾病进展的独立性危险因素,而蛋白尿的发生发展与足细胞的损伤密切相关。目的:观察移植黄芪甲苷孵育的人脂肪源性干细胞(astragaloside-incubated adipose stem cells,Ast-hADSCs)对糖尿病肾病模型大鼠肾损伤的修复作用。方法:人脂肪源性干细胞购自美国ATCC公司,雄性SD大鼠购自上海中医药大学动物实验中心,实验方案经上海中医药大学动物实验伦理委员会批准。将SD大鼠随机分为正常组、模型组、人脂肪源性干细胞组及Ast-hADSCs组,后3组应用左肾摘除联合链脲佐菌素注射法建立糖尿病肾病大鼠模型,2个干细胞干预组经尾静脉输注人脂肪源性干细胞或Ast-hADSCs悬液,2周1次,共6次,模型组予等量生理盐水。实验结束(23周)取材,检测肾功能、血脂、尿微量白蛋白及尿蛋白定量,苏木精-伊红及MASSON染色观察肾脏病理变化及肾小球面积,WT-1染色检测足细胞密度,激光共聚焦观察干细胞在肾脏的分布情况。结果与结论:①与模型组比较,2个干细胞干预组总胆固醇、尿微量白蛋白及尿蛋白定量均下降,但Ast-hADSCs组下降更显著(P<0.05);②与模型组比较,2个干细胞干预组模型大鼠的三酰甘油、低密度脂蛋白、尿素氮、高密度脂蛋白水平无统计学意义(P>0.05);③肾脏病理提示:2个干细胞干预组的肾脏损伤减轻,肾脏指数及肾小球截面积显著缩小,足细胞密度显著增加(P<0.01);Ast-hADSCs组模型大鼠肾脏病理改善明显(P<0.05);④激光共聚焦显示:干细胞干预两组模型大鼠肾组织内可见少量红色荧光表达。结果说明:经尾静脉移植人脂肪源性干细胞可提高糖尿病肾病模型大鼠足细胞的密度,减轻肾脏损伤,从而改善蛋白尿;经黄芪甲苷孵育48h后,可在一定程度上提高人脂肪源性干细胞对糖尿病肾病模型大鼠受损肾组织的修复作用。 BACKGROUND:Proteinuria is an independent risk factor for the progression of diabetic nephropathy,and the occurrence and development of proteinuria is closely related to podocyte injury.OBJECTIVE:To observe the effect of astragaloside-incubated human adipose-derived stem cells(Ast-hADSCs)on renal injury in a rat model of diabetic nephropathy.METHODS:Human adipose-derived stem cells were purchased from ATCC,USA.Male Sprague-Dawley rats were purchased from the Animal Experimental Center of Shanghai University of Traditional Chinese Medicine.The study protocol was approved by the Animal Ethics Committee of Shanghai University of Traditional Chinese Medicine.Rats were randomly divided into normal group,model group,hADSCs group and Ast-hADSC group.Animal models of diabetic nephropathy were established by left kidney extraction combined with streptozotocin injection in the latter three groups.hADSCs or Ast-hADSCs suspension was infused via the tail vein in the two intervention groups,every 2 weeks,for six sessions.Rats in the model group were given the same volume of normal saline.Levels of renal function indicators,blood lipids,urine microalbumin and urine protein were detected at 23 weeks.The pathological changes and glomerular area of the kidney were observed by hematoxylin-eosin staining and Masson staining.The podocyte density was detected by WT-1 staining.The distribution of stem cells in the kidney was observed by laser confocal microscopy.RESULTS AND CONCLUSION:Compared with the model group,there was a reduction in total cholesterol,urine microalbumin and urine protein levels decreased in the two intervention groups.Moreover,a greater reduction was found in the Ast-hADSC group than the hADSC group(P<0.05).Triacylglycerol,low-density lipoprotein,urea nitrogen,high-density lipoprotein levels were similar in the model,hADSC and Ast-hADSC groups(P>0.05).Renal pathological findings revealed a relief in renal damage and reduced renal index and glomerular cross-sectional area but significantly increased podocyte densit
作者 高俊丽 朱赟洁 刘国香 王巍巍 张金元 Gao Junli;Zhu Yunjie;Liu Guoxiang;Wang Weiwei;Zhang Jinyuan(Department of Nephrology,the 455th Hospital of PLA,Shanghai 201203,China;Department of Nephrology,Jinshan Branch,Shanghai Sixth People’s Hospital,Shanghai 201500,China)
出处 《中国组织工程研究》 CAS 北大核心 2019年第29期4629-4636,共8页 Journal of Clinical Rehabilitative Tissue Engineering Research
基金 上海市高级中西医结合人才培养计划(ZY3-RCPY-4-2039),项目负责人:王巍巍 南京军区医学科技创新项目重大课题(14ZX05).
关键词 糖尿病肾病 人脂肪源性干细胞 黄芪甲苷 PKH26 细胞移植 足细胞 蛋白尿 肾功能 diabetic nephropathy human adipose-derived stem cells astragaloside IV PKH26 cell transplantation podocytes proteinuria renal function
作者简介 高俊丽,女,1990年生,汉族,湖北省人,硕士,初级,主要从事干细胞治疗糖尿病肾病研究;通讯作者:王巍巍,博士,副主任医师,中国人民解放军第四五五医院肾脏科,上海市201203
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