Combination of acupuncture with placenta derived mesenchymal stem cell transplantation for cerebral palsy in a rat model

doi:10.3969/j.issn.2095-4344.2016.19.008

Abstract

Abstract:

BACKGROUND: Both of acupuncture and mesenchymal stem cell transplantation have therapeutic effects on cerebral palsy.
OBJECTIVE: To explore the therapentic effect of acupuncture combined with placenta derived mesenchymal stem cells on rat cerebral palsy.
METHODS: Thirty-five Wistar rats were equivalently randomized into five groups, including control group, model group, acupuncture group, transplantation group and combination group. Rat models of cerebral palsy were prepared in all groups except the control group. At 7 days after modeling, in the transplantation group, 10 μL placenta derived mesenchymal stem cells were transplanted into the right cerebral cortex and striatum of rats; rats in theacupuncture group were given acupuncture at Shuigou (DU26), Dazhui (DU14), Baihui (DU20), Guanyuan (RN4) and Qihai (RN6) points; rats in the combination group given acupuncture and transplantation treatment as described above; the model and control groups received no treatment. Then, after 7-day intervention, the body mass, neurological function and pathological changes of rat brain tissues were measured and observed, respectively. .
RESULTS AND CONCLUSION: The order of the growth rate of body mass was as follows: control group > combination group > acupuncture and transplantation groups > model group, and all groups had significant differences except the two groups of acupuncture and transplantation (P < 0.05); findings from holding tract, foot error and water finding in maze tests showed the neurological function was best in the control group, followed by the combination group, as well as acupuncture and transplantation groups, and the worst in the model group. All groups had significant differences except acupuncture and transplantation groups (P < 0.05). Besides, in the model group, cells in the brain and hippocampus decreased, arranging disorderly with fuzzy structure, and inflammatory cells appeared; pathological changes in the other groups were in between control group and model group, Additionally, brain tissues in the combination group exlibited better pathological changes than those in the acupuncture and transplantation groups, which were similar with the normal brain tissues. To conclude, acupuncture, placenta derived mesenchymal stem cells transplantation, or their combination can improve neurological function in rats with cerebral palsy, especially the combination treatment.

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: Cerebral Palsy, Acupuncture, Placenta, Mesenchymal Stem Cell Transplantation, Tissue Engineering

CLC Number:

R394.2

Shi Hua, Zhang Pu, Guo Xin, Jie Xiao-su. Combination of acupuncture with placenta derived mesenchymal stem cell transplantation for cerebral palsy in a rat model[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(19): 2789-2795.

Figures/Tables 1

2.1 胎盘间充质干细胞的形态将胎盘组织接种到培养瓶中进行原代培养，48 h见少量细胞从胎盘组织中爬出，培养第5天时出现较多的贴壁细胞，并形成细胞集落，部分出现克隆样增生，12-14 d时贴壁细胞融合达90%以上，呈漩涡状分布。传代培养后第1-3代细胞呈多样化，多为成纤维细胞，也可见到圆形或者多角形细胞，传代至第5代时，成纤维细胞为主，细胞大小一致(图1)。 2.2 胎盘间充质干细胞的鉴定流式细胞仪检测结果显示：第4代细胞CD29、CD44、CD90和CD105细胞表型呈阳性表达，CD45细胞表型呈阴性表达。 2.3 实验动物数量分析 35只大鼠依从性良好，无脱落，均进入分析。 2.4 各组大鼠的体质量增长情况对照组、模型组、针刺组、移植组和针刺联合移植组大鼠体质量增长率分别为(99.61±8.36)%，(59.17±7.63)%，(76.54± 14.11)%，(78.37±16.15)%，(83.43±7.75)%。统计学分析显示，模型组大鼠的体质量比对照组大鼠明显减轻，两者比较差异有显著性意义(P < 0.05)，针刺组、移植组和针刺联合移植组大鼠的体质量均较对照组轻(P < 0.05)、较模型组重(P < 0.05)，针刺组和移植组之间比较差异无显著性意义(P > 0.05)，针刺联合移植组大鼠的体质量较针刺组和移植组增加(P < 0.05)。 2.5 各组大鼠握持牵引实验结果对照组、模型组、针刺组、移植组、针刺联合移植组大鼠的握持牵引时间分别为(46.51±12.04) s，(29.13±12.53) s，(35.53±13.51) s，(36.25±17.24) s，(43.13±9.27) s。对照组的握持时间最长，模型组的握持时间最短，针刺组、移植组和针刺联合移植组的握持时间介于两者之间，针刺组、移植组和针刺联合移植组的握持时间与模型组和对照组比较，差异均有显著性意义(P < 0.05)，针刺组和移植组的握持时间比较，差异无显著性意义(P > 0.05)，针刺联合移植组的握持时间大于针刺组和移植组，差异均有显著性意义(P < 0.05)。 2.6 各组大鼠足错误实验结果对照组、模型组、针刺组、移植组、针刺联合移植组大鼠的足错误次数分别为0.69±0.71，3.54±1.33，2.27±1.32，2.32±0.23，1.14±0.67。对照组的错误次数最少，模型组的错误次数最多，针刺组、移植组和针刺联合移植组的错误次数介于两者之间，针刺组、移植组和针刺联合移植组的错误次数与模型组和对照组比较，差异均有显著性意义(P < 0.05)，针刺组和移植组的错误次数比较，差异无显著性意义(P > 0.05)，针刺组联合移植组的错误次数少于针刺组和移植组的错误次数，差异均有显著性意义(P < 0.05)。 2.7 各组大鼠的迷宫觅水实验结果 2.7.1 觅水时间结果对照组、模型组、针刺组、移植组、针刺联合移植组大鼠的觅水时间分别为(30.11± 8.14) s，(200.14±22.73) s，(165.57±33.33) s，(166.45± 27.25) s，(123.31±11.47) s。对照组的觅水时间最短，模型组的觅水时间最长，针刺组、移植组和针刺联合移植组的觅水时间介于两者之间，针刺组、移植组和针刺联合移植组的觅水时间与模型组和对照组比较，差异均有显著性意义(P < 0.05)，针刺组和移植组的觅水时间比较，差异无显著性意义(P > 0.05)，针刺组联合移植组的觅水时间短于针刺组和移植组，差异均有显著性意义(P < 0.05)。 2.7.2 觅水错误次数结果对照组、模型组、针刺组、移植组、针刺联合移植组大鼠的觅水错误次数分别为7.66±1.42，13.43±1.35，11.23±2.31，11.35± 1.42)，9.52±1.73。对照组的觅水错误次数最少，模型组的觅水错误次数最多，针刺组、移植组和针刺联合移植组的觅水错误次数介于两者之间，针刺组、移植组和针刺联合移植组的觅水错误次数与模型组和对照组比较，差异均有显著性意义(P < 0.05)，针刺组和移植组的觅水错误次数比较，差异无显著性意义(P > 0.05)，针刺联合移植组的觅水错误次数少于针刺组和移植组，差异均有显著性意义(P < 0.05)。 2.7.3 觅水重复次数结果对照组、模型组、针刺组、移植组、针刺联合移植组大鼠的觅水重复次数分别为1.55±0.73，3.47±0.88，2.18±0.54，2.19±0.75，1.87±0.60，对照组的觅水重复次数最少，模型组的觅水重复次数最多，针刺组、移植组和针刺联合移植组的觅水重复次数介于两者之间，针刺组、移植组和针刺联合移植组的觅水重复次数与模型组和对照组比较，差异均有显著性意义(P < 0.05)，针刺组和移植组的觅水重复次数比较，差异无显著性意义(P > 0.05)，针刺联合移植组的觅水重复次数少于针刺组和移植组，差异均有显著性意义(P < 0.05)。 2.8 各组大鼠脑组织的病理变化各组大鼠脑组织苏木精-伊红染色结果显示：对照组大鼠大脑组织和海马区脑组织的细胞着色深，排列整齐，层次清楚，形态和结构正常，模型组大鼠的大脑组织和海马区脑组织的细胞数量减少，排列紊乱，见囊性变，有炎性细胞聚集，细胞结构模糊。针刺组、移植组和针刺联合移植组大脑和海马组织病理改变介入对照组和模型组之间，其中针刺联合移植组的病理改善较针刺组和移植组好，有向正常大脑组织接近的趋势，见图2。"

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