Bone marrow mesenchymal stem cells protect against renal ischemia-reperfusion injury through immune regulatory mechanism

doi:10.3969/j.issn.2095-4344.2014.37.013

Abstract

Abstract:

BACKGROUND: Stem cell therapy for renal ischemia-reperfusion injury has been the hot topics for many scholars. Its mechanism is very complex, which could not be explained by simple mechanism of stem cells differentiation. It is the result involving a variety of mechanisms.
OBJECTIVE: To investigate the influence on immune cells during the bone marrow mesenchymal stem cell therapy for renal ischemia-reperfusion injury, then to preliminary summarize the immune regulation mechanism of bone marrow mesenchymal stem cell therapy for renal ischemia-reperfusion injury.
METHODS: First, we established a model of renal ischemia-reperfusion injury in rats and, cultured and purified rat bone marrow mesenchymal stem cells in vitro. Then, the bone marrow mesenchymal stem cells were transplanted into the rat models. Using flow cytometry detection technology, we analyzed the proportion of CD4⁺CD25⁺regulatory T cells of rat spleen cells, discussed the effects on immune cells during the bone marrow mesenchymal stem cell therapy for renal ischemia-reperfusion injury, and then transferred the rat’s spleen cells to the nude mice which were subjected to renal renal ischemia-reperfusion injury. Renal function and renal histological changes of nude mice were assessed.
RESULTS AND CONCLUSION: The bone marrow mesenchymal stem cell transplantation could significantly inhibit the decrease of CD4⁺CD25⁺ regulatory T cell of spleen cells in rats with renal ischemia-reperfusion injury. The transplantation of spleen cells from the above-mentioned rats to nude mice could obviously protect nude mice from renal ischemia-reperfusion injury, characterized by lower serum creatinine, blood urea nitrogen and renal tubule pathologic damage score. Therefore, bone marrow mesenchymal stem cells have protective effects on renal ischemia-reperfusion injury by regulating the immune system.

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

全文链接：

Key words: bone marrow, mesenchymal stem cells, kidney, reperfusion injury, T-lymphocytes

CLC Number:

R394.2

Hu Hong-lin, Zou Cong, Xi Xiao-qing, Ye Zhen-feng, Jiang Wei . Bone marrow mesenchymal stem cells protect against renal ischemia-reperfusion injury through immune regulatory mechanism[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(37): 5977-5982.

Figures/Tables 1

2.1 实验动物数量分析 18只SD大鼠，骨髓间充质干细胞移植组有1只术后5 h死亡，15只裸鼠，脾细胞移植组和对照组各有1只术后8 h和6 h死亡，其余动物全部存活。 2.2 骨髓间充质干细胞生长特点骨髓单个核细胞接种于培养瓶后，细胞呈圆形，透亮，72 h换液去除未贴壁细胞后，可见散在的贴壁细胞呈长梭形，第3天后细胞增殖加快，贴壁细胞增多，呈成纤维样细胞形态，细胞扁平无立体感。10-14 d细胞铺满瓶底，达到融合状态。传代培养的骨髓间充质干细胞呈圆形，24 h内完全贴壁，形态与原代培养的细胞类似，生长迅速，1周左右达到完全融合。传至8-10代以后，细胞分裂增殖速度明显减慢，细胞内颗粒增多，细胞逐渐老化。体外传代培养4-6代的骨髓间充质干细胞经锥虫蓝染色显示细胞活性大于95%。 2.3 骨髓间充质干细胞移植对肾缺血再灌注损伤大鼠脾细胞CD4+CD25+调节性T细胞亚型的影响流式细胞结果显示，正常组、对照组、骨髓间充质干细胞移植组大鼠脾脏中CD4+CD25+调节性T细胞的比例分别为(12.81± 2.76)%，(9.21±1.79)%，(16.12±2.34)%。与正常组比较，对照组大鼠脾脏中CD4+CD25+调节性T细胞的比例明显降低(P < 0.05)；与对照组比较，骨髓间充质干细胞移植组大鼠脾脏中CD4+CD25+调节性T细胞的比例明显升高(P < 0.05)，见图1。 2.4 移植大鼠脾细胞对裸鼠肾缺血再灌注损伤后肾功能的影响裸鼠经历45 min双侧肾脏缺血，再灌注后48 h各组血清肌酐和尿素氮水平见图2。与正常组相比，对照组血清肌酐和尿素氮均明显升高，差异有显著性意义(P < 0.05)；与对照组相比，脾细胞移植组血清肌酐和尿素氮均明显降低，差异有显著性意义(P < 0.05)。 2.5 移植大鼠脾细胞对裸鼠肾缺血再灌注损伤后肾脏组织学的影响正常组肾脏形态、大小、色泽正常；而缺血组肾脏稍肿大，髓质可见轻重不一的淤血暗红。苏木精-伊红染色光镜检查显示：正常组肾脏组织结构基本正常(图3A)；对照组肾组织见大量小管上皮细胞肿胀、空泡变性，部分小管上皮细胞刷状缘扁平、皱缩、核深染、裸露。严重者可见上皮细胞坏死、崩解脱落以及细胞核浓缩、碎裂、核溶解。部分肾小管管腔扩张，坏死、崩解的上皮细胞可脱落入肾小管管腔内，呈模糊的颗粒样结构；肾间质见炎性细胞浸润(图3B)。脾细胞移植组肾小管上皮细胞坏死明显减轻(图3C)。肾小管病理损伤评分结果所示，对照组病理评分均显著高于正常组，差异有显著性意义(P < 0.01)，而脾细胞移植组病理评分显著低于对照组，差异有显著性意义 (P < 0.05)，见图4。"

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