Effect of corbrin capsule combined with human amniotic mesenchymal stem cell transplantation on rat renal function and hypercoagulability

doi:10.3969/j.issn.2095-4344.2017.01.024

Abstract

Abstract:

BACKGROUND: As the main component of cordycepic acid, corbrin capsule mainly functions to improve blood lipids and protect the kidneys. Human amniotic mesenchymal stem cells are a kind of stem cells that have stronger differentiation potential than bone marrow mesenchymal stem cells. However, there are less reports on renal injury repair and post-repair functional improvement.
OBJECTIVE: To observe the effect of human amniotic mesenchymal stem cell transplantation with corbrin capsule on blood coagulation and renal functions of rats with nephrotic syndrome.
METHODS: Human amniotic mesenchymal stem cells were cultured in vitro and made into cell suspension labeled with with CM-Dil. Fifty Sprague-Dawley were randomly divided into five groups (n=10 per group): normal control group with no intervention, model group, cell transplantation group, corbrin capsule group and combined treatment group. Except the normal control groups, rats in all the groups were used to make nephropathy models by rats by intravenous injection of adriamycin. After modeling, rats in the latter three groups were given intravenous injection of human amniotic mesenchymal stem cells for 3 days, intragastric administration of corbrin capsule for 12 weeks, and the combination of cell transplantation and corbrin capsule, respectively. Twelve weeks later, serum creatinine, blood urea nitrogen, blood coagulation index, and 24-hour urine protein levels were detected. Pathological changes of the renal tissues were observed by hematoxylin-eosin staining. Survival and distribution of CM-Dil labeled human amniotic mesenchymal stem cells were observed by fluorescence microscope. Apoptosis in renal tissues was detected using TUNEL method.
RESULTS AND CONCLUSION: Serum creatinine, blood urea nitrogen and 24-hour urine protein levels in the cell transplantation and corbrin capsule groups were significantly decreased compared with the model group, but still higher than that in the combined treatment group (P < 0.05). Coagulation indexes in the combined treatment group were significantly lower than those in the cell transplantation and corbrin capsule groups (P < 0.05). The hypercoagulable state of rats was relieved in the three treatment groups. Pathological changes of the renal tissues were deteriorated in the model group, and improved in the three treatment groups, especially in the combined treatment group. Under the florescence microscope, the number of CM-Dil-labeled human amniotic mesenchymal stem cells was higher in the combined treatment group than the cell transplantation and corbrin capsule groups. Compared with the model group, the cell apoptosis in the renal tissue was relieved in the three treatment groups, especially in the combined treatment (P < 0.05). To conclude, human amniotic mesenchymal stem cell transplantation with corbrin capsule can improve renal function and hypercoagulable state of nephrotic syndrome rats, by reducing the degree of proteinuria, improving the degree of pathological damage and reducing renal cell apoptosis.

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: Stem Cells, Amnion, Mesenchymal Stem Cells, Nephrosis, Lipoid, Tissue Engineering

CLC Number:

R394.2

Guo Zhi-bo, Zhang Chen-jie, Ma Li-na, Gao Da-wei, Zhou Fang, Li Na-na, Zhou Jun-xue, Zhang Na. Effect of corbrin capsule combined with human amniotic mesenchymal stem cell transplantation on rat renal function and hypercoagulability[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(1): 133-139.

Figures/Tables 2

2.1 实验动物数量分析纳入50只大鼠，分为5组，取10只为正常组，余40只大鼠随机分为4组，全部进入结果分析，无脱失。 2.2 人羊膜间充质干细胞的形态观察人羊膜间充质干细胞复苏后在体外培养五至六天可传代，传代后24 h内即可呈贴壁生长，形态多样，呈梭形、纺锤形、星形或多边形。 P2代的人羊膜间充质干细胞贴壁速度加快，呈长梭形集落生长，均一性较好，显微镜下可见胞浆含有中间丝及高尔基复合体等。P3的人羊膜间充质干细胞形态单一，纯度较好，细胞排列紧密，呈漩涡状分布。第3代的人羊膜间充质干细胞见图1。 2.3 各组大鼠血肌酐、尿素氮、凝血指标、24 h尿蛋白水平比较人羊膜间充质干细胞组、百令胶囊组血肌酐和尿素氮、24 h尿蛋白水平均高于联合组，但明显低于模型组，大鼠的血液高凝状态有所缓解，差异有显著性意义(P < 0.05)；联合组血生化指标低于人羊膜间充质干细胞组、百令胶囊组(P < 0.05)。各组大鼠血肌酐、尿素氮、24 h尿蛋白水平检测结果见表1及图2，各组大鼠凝血指标检测结果见表2及图3。 2.4 肾组织病理学变化苏木精-伊红染色结果显示，联合组的病理改善情况优于人羊膜间充质干细胞组和百令胶囊组，模型组的肾脏病理没有改善甚至恶化，人羊膜间充质干细胞组和百令胶囊组稍有改善，但不及联合组。各组大鼠的病理学变化比较见图4。 2.5 CM-Dil标记羊膜间充质干细胞存活及分布情况荧光显微镜观测CM-Dil标记人羊膜间充质干细胞的存活情况，与人羊膜间充质干细胞组比较，联合组CM-Dil标记的人羊膜间充质干细胞明显增多，见图5。 2.6 各组大鼠肾脏细胞凋亡情况模型组阳性细胞凋亡面积最大，人羊膜间充质干细胞组、百令胶囊组可见阳性细胞凋亡，联合组细胞凋亡面积较小，组间比较差异均有显著性意义(P < 0.05)，见表3及图6。"

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