Co-treatment of endothelial progenitor cells and pioglitazone improves kidney function in diabetic rats

doi:10.3969/j.issn.2095-4344.2017.17.013

Abstract

Abstract:

BACKGROUND: Pioglitazone is a common hypoglycemic drug capable of improving proliferation and activation, and inhibiting apoptosis of endothelial progenitor cells (EPCs). We speculated that the combined use of pioglitazone and EPCs transplantation could have significant improving effects on hyperglycemia and kidney function after diabetes mellitus.
OBJECTIVE: To investigate the improving effect of EPCs transplantation combined with pioglitazone treatment on the kidney function of diabetic rat models.
METHODS: The 15 of 75 Wistar rats were randomly selected and served as normal control group (no treatment). Animal models of type 1 diabetes mellitus were made in the rest 60 rats through the intraperitoneal injection of 40 mg/kg streptozotocin for continuous 5 days. The human EPCs (labeled by CM-Dil) were recovered, cultured and preserved until transplantation. After 4 weeks of modeling, the 60 rat models were randomly divided into model group (PBS injection), pioglitazone group, EPCs transplantation group and combined treatment group, followed by tail vein injection of EPCs suspension and/or intragastric administration of 20 mg/kg pioglitazone for continuous 4 days. After 8 weeks of treatment, the levels of glucose, insulin and creatinine in serum, urea nitrogen and urine protein during 24 hours were determined. The number and distribution of EPCs labeled by CM-Dil were detected by fluorescence microscope, the apoptosis in kidney cells was tested by TUNEL method, and the kidney weight/body weight ratio in rats was calculated.
RESULTS AND CONCLUSION: Compared with the model group, the blood glucose and serum creatinine levels and the urea nitrogen and urine protein concentrations during 24 hours were significantly decreased (P < 0.05), and the serum insulin level was significantly increased (P < 0.05) in the pioglitazone and EPCs transplantation groups. These biochemical indexes in the combined treatment group were more significantly altered compared with the model group (P < 0.01). The kidney weight to the body weight ratio was lowest in the combined treatment group and lower in the pioglitazone and EPCs transplantation groups followed by the model group (P < 0.05). The order of apoptotic kidney cells labeled by TUNEL was as follows: model group > pioglitazone group > EPCs transplantation group > combined treatment group (P < 0.05). To conclude, the EPCs transplantation combined with pioglitazone treatment can decrease the blood glucose and serum creatinine levels and urea nitrogen and urine protein concentrations, improve the serum insulin level, reduce cell apoptosis in the kidney, and remit the kidney dysfunction of diabetic rats to a certain extent.

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

Key words: Endothelial Cells, Thiazolidinediones, Cell Transplantation, Diabetic Nephropathies, Tissue Engineering

CLC Number:

R394.2

Wang Song, Zheng Xi-sheng, Wang Shan, Fang Fang. Co-treatment of endothelial progenitor cells and pioglitazone improves kidney function in diabetic rats[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(17): 2702-2707.

Figures/Tables 1

2.1 内皮祖细胞形态接种后4-6 d，部分细胞开始贴壁增殖，随着时间延长贴壁细胞明显增多并且体积变大，细胞呈现类圆形、纺锤形或多边形，见图1A；7-14 d时，贴壁细胞开始膨胀，呈现为大圆形，部分贴壁细胞生长呈“铺路石”样外观，见图1B。当细胞满铺80%时进行传代。细胞复苏后存活率大约85%。复苏传代后的细胞形态与未冻存的细胞无明显差异。 2.2 各组大鼠血糖、胰岛素、血肌酐、尿素氮?24 h尿蛋白水平如表1所示，与正常对照组相比，糖尿病组血糖、尿素氮、血肌酐、24 h尿蛋白含量均显著升高，而胰岛素含量则显著降低。与糖尿病模型组比较，内皮祖细胞组、吡格列酮组大鼠血糖、尿素氮、血肌酐、24 h尿蛋白水平明显降低，血清胰岛素含量明显升高，差异有显著性意义(P < 0.05)；联合治疗组大鼠血糖、尿素氮、血肌酐、24 h尿蛋白水平降低更明显(P < 0.01)，血清胰岛素浓度升高更明显(P < 0.01)；尽管联合治疗组大鼠的生化指标仍未得到完全恢复，并与正常对照组相比差异仍有显著性意义，但可见恢复趋势。 2.3 大鼠体质量?肾质量变化与其他各组相比，正常对照组肾质量/体质量比值最低(P < 0.05)。与糖尿病模型组大鼠相比，内皮祖细胞组、吡格列酮组、联合治疗组显著降低(P < 0.05)；联合治疗组肾质量/体质量比值明显低于内皮祖细胞组和吡格列酮组(P < 0.05)，见表2。 2.4 CM-Dil标记的内皮祖细胞存活及分布情况治疗8周后，正常对照组、糖尿病模型组和吡格列酮组未见有CM-Dil标记阳性细胞。内皮祖细胞移植组和联合治疗组大鼠肾组织中可见明显的CM-Dil标记的内皮祖细胞；联合治疗组大鼠CM-Dil标记阳性细胞明显多于内皮祖细胞移植组，见图2。 2.5 各组肾组织细胞凋亡情况如表3、图3所示，正常对照组细胞凋亡数最少，糖尿病模型组细胞凋亡数最多，与糖尿病模型组比较，内皮祖细胞移植组和吡格列酮组凋亡细胞数明显减少(P < 0.05)，联合治疗组进一步减少(P < 0.01)。"

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