人脐带间充质干细胞对糖尿病肾病大鼠肠道屏障功能的调节作用  

Regulatory effect of human umbilical cord mesenchymal stem cells on intestinal barrier function in diabetic nephropathy rats

作　　者：吴雅茹 米焱[2] 魏凯悦 高和平 张丁予 王彩丽[2] Wu Yaru;Mi Yan;Wei Kaiyue;Gao Heping;Zhang Dingyu;Wang Caili(Baotou Medical College of Inner Mongolia University of Science and Technology,Baotou 014000,Inner Mongolia Autonomous Region,China;Department of Nephrology,First Affiliated Hospital of Baotou Medical College,Inner Mongolia University of Science and Technology,Baotou 014000,Inner Mongolia Autonomous Region,China)

机构地区：[1]内蒙古科技大学包头医学院,内蒙古自治区包头市014000 [2]内蒙古科技大学包头医学院第一附属医院肾内科,内蒙古自治区包头市014000

出　　处：《中国组织工程研究》2024年第19期2967-2973,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(82160141),项目负责人:王彩丽。

摘　　要：背景:糖尿病肾病是引起终末期肾病的重要原因,其中肠道屏障损伤在糖尿病肾病的发生发展中起到了重要作用。目的:观察人脐带间充质干细胞对糖尿病肾病大鼠肠道屏障的保护作用。方法:30只8周龄雄性SD大鼠随机分配至健康对照组、模型组和人脐带间充质干细胞组,每组10只。人脐带间充质干细胞组大鼠在糖尿病肾病成模后经尾静脉注射1×106个人脐带间充质干细胞,每周1次,连续4周,健康对照组和模型组大鼠在相同时间注射等体积PBS。末次注射后1周,光镜下观察肾脏和结肠组织形态学改变情况,免疫组化法检测大鼠结肠组织中ZO-1和Occludin的表达;ELISA法检测大鼠血清D-乳酸和脂多糖水平。此外,还利用活体成像系统观察DiR染料标记的人脐带间充质干细胞在大鼠体内的示踪分布,以及应用免疫组化法检测结肠组织中人源性间充质干细胞表面标志抗原CD44和CD90的表达。结果与结论:①与模型组比较,人脐带间充质干细胞移植显著抑制了糖尿病肾病大鼠尿素氮、血肌酐、24 h尿蛋白水平和尿白蛋白/肌酐比值的增加(均P<0.05);②在糖尿病肾病大鼠结肠中发现了人源间充质干细胞表面标志物CD44和CD90的表达;③与健康对照组相比,模型组大鼠结肠组织中紧密连接蛋白Occludin与ZO-1蛋白表达水平显著减少,而人脐带间充质干细胞治疗后显著提高了Occludin和ZO-1的表达;④与模型组比较,人脐带间充质干细胞移植后显著降低了糖尿病肾病大鼠血清D-乳酸和脂多糖水平;⑤结果提示,人脐带间充质干细胞可能通过增强糖尿病肾病大鼠肠道紧密连接蛋白的表达来保护肠道屏障功能。BACKGROUND:Diabetic nephropathy is an important cause of end-stage renal disease,and intestinal barrier damage plays an important role in the occurrence and development of diabetic nephropathy.OBJECTIVE:To observe the protective effect of human umbilical cord mesenchymal stem cells on the intestinal barrier in rats with diabetic nephropathy.METHODS:Thirty 8-week-old male SD rats were randomly assigned to healthy control group,model group and human umbilical cord mesenchymal stem cell group,with 10 rats in each group.Rats in the human umbilical cord mesenchymal stem cell group were injected with 1×106 human umbilical cord mesenchymal stem cells through the tail vein once a week for 4 weeks after the model establishment of diabetic nephropathy.Rats in the healthy control group and the model group were injected with an equal volume of PBS at the same time.1 week after the last injection,the histomorphological changes in the kidney and colon were observed under a light microscope.The expressions of ZO-1 and Occludin in the colon tissue of rats were detected by immunohistochemistry.Serum D-lactic acid and lipopolysaccharide levels were detected by ELISA.In addition,the distribution of human umbilical cord mesenchymal stem cells labeled with DiR dye in rats was observed by in vivo imaging system.The expression of human mesenchymal stem cell surface marker antigens CD44 and CD90 in colon tissue was detected by immunohistochemistry.RESULTS AND CONCLUSION:(1)Compared with the model group,human umbilical cord mesenchymal stem cell transplantation significantly inhibited the increase of urea nitrogen,serum creatinine,24-hour urine protein level and urinary albumin/creatinine ratio in diabetic nephropathy rats(all P<0.05).(2)The expression of human mesenchymal stem cell surface markers CD44 and CD90 was found in the colon of diabetic nephropathy rats.(3)Compared with the healthy control group,the expression levels of tight junction proteins Occludin and ZO-1 in the colon tissue of the model group were significantly reduced,

关 键 词：人脐带间充质干细胞 糖尿病肾病 肠道屏障 ZO-1 OCCLUDIN 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R692.6