Chinese Journal of Tissue Engineering Research ›› 2014, Vol. 18 ›› Issue (19): 2999-3004.doi: 10.3969/j.issn.2095-4344.2014.19.008
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Li Yan-ju1, Liu Yang2, Wang Ji-shi1, Meng Wei3, Huang Yi1
Revised:
2014-04-17
Online:
2014-05-07
Published:
2014-05-07
Contact:
Li Yan-ju, Department of Hematology, Affiliated Hospital of Guiyang Medical College, Guiyang 550004, Guizhou Province, China
About author:
Li Yan-ju, M.D., Associate chief physician, Department of Hematology, Affiliated Hospital of Guiyang Medical College, Guiyang 550004, Guizhou Province, China
Supported by:
the Science and Technology Plan Fund of Guizhou Province, No. SY[2010]3075; the Grant from the Health Bureau of Guizhou Province, No. gzwkj2011-1-023
CLC Number:
Li Yan-ju, Liu Yang, Wang Ji-shi, Meng Wei, Huang Yi. Effect and mechanism of mesenchymal stem cells on the aging kidney[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(19): 2999-3004.
2.1 实验动物数量分析 大鼠30只实验中无脱落现象,均进入统计结果分析。 2.2 骨髓间充质干细胞培养及流式细胞仪鉴定的结果 实验采用贴壁法培养的骨髓间充质干细胞光学显微镜下见细胞呈星形、梭形,见图1A,B。 经流式细胞仪检测,结果显示骨髓间充质干细胞的特异性表面标志物:CD45阴性,阳性率为0.83%,CD44阳性,阳性率为92.16%,CD29阳性,阳性率为91.23%。提示培养的细胞主要是骨髓间充质干细胞,见图1D-G。 2.3 骨髓间充质干细胞在肾脏组织的归巢 CFSE为一种绿色荧光染料,可以穿透细胞膜与细胞核中的双链DNA结合而发挥标记的作用,并对细胞的生理活动没有影响,是一种良好的细胞标记物。实验将终浓度为20 μmol/L CFSE染色骨髓间充质干细胞输注模型组大鼠体内24 h后,取出肾脏组织制作冰冻切片,荧光显微镜下观察有荧光细胞存在,见图1C。"
2.4 骨髓间充质干细胞移植后大鼠肾脏组织超氧化物歧酶和丙二醛表达 分别于骨髓间充质干细胞细胞移植后第4周处死大鼠,检测正常组肾脏组织中超氧化物歧酶的活性和丙二醛含量分别为:(31.26±2.24) U/mg,(2.756± 0.128) μmol/g;模型组肾脏组织中超氧化物歧酶的活性为(11.47±2.32) U/mg,低于正常组(P < 0.05),模型组丙二醛含量为(5.367±0.682) μmol/g,高于正常组(P < 0.05),说明肾脏衰老模型制作成功。 2.5 骨髓间充质干细胞移植后大鼠肾脏组织尿素氮及肌酐的浓度变化 分别于骨髓间充质干细胞细胞移植后第4周处死大鼠,模型组较正常组的尿素氮及肌酐相比呈升高趋势。治疗组尿素氮及肌酐低于模型组(P < 0.05),见表1。"
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