中国组织工程研究

中国组织工程研究 ›› 2019, Vol. 23 ›› Issue (29): 4629-4636.doi: 10.3969/j.issn.2095-4344.1799

• 脂肪干细胞 adipose-derived stem cells • 上一篇    下一篇

黄芪甲苷孵育的脂肪干细胞治疗糖尿病肾病

高俊丽1，2，朱赟洁1，刘国香1，王巍巍1，张金元1   

  1. 1中国人民解放军第四五五医院肾脏科，上海市  201203；2上海市第六人民医院金山分院肾内科，上海市  201500
  • 修回日期:2019-03-20 出版日期:2019-10-18 发布日期:2019-10-18
  • 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
  • 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。
  • 基金资助:

    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)，项目负责人：王巍巍

Astragaloside-incubated adipose-derived stem cells for the treatment of diabetic nephropathy

Gao Junli1, 2, Zhu Yunjie1, Liu Guoxiang1, Wang Weiwei1, Zhang Jinyuan1   

  1. 1Department of Nephrology, the 455th Hospital of PLA, Shanghai 201203, China; 2Department of Nephrology, Jinshan Branch, Shanghai Sixth People’s Hospital, Shanghai 201500, China
  • Revised:2019-03-20 Online:2019-10-18 Published:2019-10-18
  • Contact: Wang Weiwei, MD, Associate chief physician, Department of Nephrology, the 455th Hospital of PLA, Shanghai 201203, China
  • About author:Gao Junli, Master, Department of Nephrology, the 455th Hospital of PLA, Shanghai 201203, China; Department of Nephrology, Jinshan Branch, Shanghai Sixth People’s Hospital, Shanghai 201500, China
  • Supported by:

    Shanghai Advanced Chinese and Western Medicine Talents Training Program, No. ZY3-RCPY-4-2039 (to WWW); Medical Science and Technology Innovation Project of the Nanjing Military Region, No. 14ZX05 (to WWW)

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摘要：

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文题释义：
黄芪甲苷：是中药黄芪的主要提取物，也是黄芪的主要成分之一。黄芪甲苷可影响干细胞增殖、抑制凋亡并诱导其分化。黄芪甲苷孵育的脂肪干细胞可增强其增殖能力，并增强其旁分泌效应，提高向受损组织迁徙能力，可提高其治疗效果。
黄芪甲苷孵育脂肪间充质干细胞的作用：作者前期的研究发现，黄芪甲苷可增强脂肪间充质干细胞的增殖能力，而不影响其分化能力，与顺铂诱导的肾小管上皮细胞共同培养，脂肪间充质干细胞向受损部位定向迁移的能力增强，能够减少肾小管上皮细胞的凋亡。经黄芪甲苷孵育后的脂肪间充质干细胞可增强其向受损组织迁徙的细胞量和存活率，提高其对受损组织的修复能力。

 

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48 h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。


中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程
ORCID: 0000-0002-4706-2490(高俊丽)

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

Abstract:

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 density in the hADSC and Ast-hADSC groups (P < 0.01). The renal pathological changes of the Ast-hADSC group were superior to those of the hADSC group (P < 0.05). Under the laser confocal microscope, a small amount of red fluorescence was expressed in the kidney. These results indicate that transplantation of human adipose-derived stem cells through the tail vein can increase the density of podocytes, reduce renal damage, and improve proteinuria in diabetic nephropathy rats. Incubation with astragaloside IV for 48 hours can promote the efficacy of human adipose-derived stem cells in the repair of damaged kidney tissue in diabetic nephropathy rats.

Key words: diabetic nephropathy, human adipose-derived stem cells, astragaloside IV, PKH26, cell transplantation, podocytes, proteinuria, renal function

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