鹿茸多肽联合GDNF基因修饰的雪旺细胞对Aβ25-35诱导脊髓神经元凋亡的保护作用

doi:10.13481/j.1671-587x.20180614

吉林大学学报(医学版) ›› 2018, Vol. 44 ›› Issue (06): 1190-1193.doi: 10.13481/j.1671-587x.20180614

鹿茸多肽联合GDNF基因修饰的雪旺细胞对Aβ_25-35诱导脊髓神经元凋亡的保护作用

张鹏程¹, 冷向阳¹, 路博丞¹, 张远超², 王英², 王旭凯²

1. 长春中医药大学临床医学院骨伤科, 吉林长春 130021;
2. 长春中医药大学附属医院骨伤科, 吉林长春 130021

收稿日期:2018-01-29 出版日期:2018-11-28 发布日期:2018-11-28
通讯作者: 王旭凯,副教授,硕士研究生导师(Tel:0431-85656164,E-mail:wangxukai_2003@163.com) E-mail:wangxukai_2003@163.com
作者简介:张鹏程(1990-),男,吉林省长春市人,在读医学博士,主要从事脊柱疾病基础和临床方面的研究。
基金资助:
吉林省科技厅科技发展计划项目资助课题（20150520049JH）

Protective effect of Schwann cells modified by VAP combined with GDNF gene on apoptosis of spinal cord neurons induced by Aβ_25-35

ZHANG Pengcheng¹, LENG Xiangyang¹, LU Bocheng¹, ZHANG Yuanchao², WANG Ying², WANG Xukai²

1. Department of Orthopedics, School of Clinical Medicine, Changchun University of Traditional Chinese Medicine, Changchun 130021, China;
2. Department of Orthopedics, Affiliated Hospital, Changchun University of Traditional Chinese Medicine, Changchun 130021, China

Received:2018-01-29 Online:2018-11-28 Published:2018-11-28

摘要/Abstract

摘要： 目的：探讨鹿茸多肽（VAP）联合胶质细胞源性神经营养因子（GDNF）基因修饰的雪旺细胞（SCs）对β淀粉样蛋白25-35（Aβ_25-35）诱导的脊髓神经元凋亡的保护作用，阐明其作用机制。方法：制备胎鼠脊髓细胞，取对数生长期的脊髓神经元，采用Aβ_25-35诱导脊髓神经元凋亡，将脊髓神经元分为正常细胞组（正常脊髓神经元）、诱导凋亡组（Aβ_25-35诱导凋亡后的脊髓神经元）、SCs组（Aβ_25-35诱导凋亡后的脊髓神经元+SCs）、GDNF组（Aβ_25-35诱导凋亡后的脊髓神经元+GDNF）、SCs+GDNF组（Aβ_25-35诱导凋亡后的脊髓神经元+GDNF转染的SCs）和VAP联合组（Aβ_25-35诱导凋亡后的脊髓神经元+VAP联合GDNF转染的SCs）。流式细胞术检测各组脊髓神经元凋亡率，免疫组织化学染色检测各组脊髓神经元中caspase-3阳性细胞数。结果：胎鼠脊髓神经元悬液接种后初始脊髓神经元大多为圆形。流式细胞术检测，与诱导凋亡组比较，SCs组、GDNF组、SCs+GDNF组和VAP联合组脊髓神经元凋亡率降低（P<0.05）；与SCs+GDNF组比较，VAP联合组脊髓神经元凋亡率明显降低（P<0.05）。免疫组织化学检测，各组脊髓神经元中均有caspase-3表达，SCs组、GDNF组和SCs+GDNF组细胞着色差异不明显，脊髓神经元中caspase-3阳性细胞数差异不大，但与诱导凋亡组比较明显减少（P<0.05）。结论：VAP联合GDNF转染的SCs对脊髓神经元凋亡有保护作用，该作用通过下调脊髓神经元中caspase-3表达来实现。

关键词: 鹿茸多肽, 胶质细胞源性神经营养因子, 雪旺细胞, 脊髓神经元

Abstract: Objective: To discuss the protective effect of velvet antler polypeptides(VAP)combined with Schwann cells(SCs) modified by glial cell line derived neurotrophic factor(GDNF)gene on the apoptosis of spinal cord neurons induced by beta-amyloid 25-35(Aβ_25-35).Methods: The spinal cord cells of fetal mice were prepared and the spinal cord neurons in logarithmic growth phase were taken; the apoptosis of spinal cord neurons was induced by Aβ_25-35.The spinal cord neurons were divided into normal cell group (the normal spinal cord neurons), induced apoptosis group (the apoptotic spinal cord neurons induced by Aβ_25-35, SCs group (the apoptotic spinal cord neurons induced by Aβ_25-35+SCs), GDNF group (the apoptotic spinal cord neurons induced by Aβ_25-35+GDNF), SCs + GDNF group (the apoptotic spinal cord neurons induced by Aβ_25-35+GDNF-transfected SCs) and VAP combination group (the apoptotic spinal cord neurons induced by Aβ_25-35+VAP combined with GDNF-transfected SCs).Flow cytometry was used to detect the apoptotic rates of spinal cord neurons in various groups; immunohistochemical staining was used to detect the number of caspase-3 positive cells in the spinal cord neurons in various groups.Results: After suspension inoculation of fetal spinal cord neurons, most of them were round at the initial stage.The flow cytometry results showed that compared with induced apoptosis group, the apoptotic rates of spinal cord neurons in SCs,GDNF,SCs+GDNF,and VAP combination groups were decreased (P<0.05). Compared with SCs+GDNF group,the apoptotic rate of spinal cord neurons in VAP combination group was decreased (P<0.05).The immunohistochemistry results showed that the expression of caspase-3 in spinal cord neurons in various groups could be found. There were no significant differences in the number of caspase-3 positive cells and cell staining between SCs group, GDNF group and SCs+GDNF group; but the number of caspase-3 positive cells in SCs group, GDNF group and SCs+GDNF group were significantly higher than that in induced apoptosis group (P<0.05).Conclusion: VAP combined with GDNF-transfected SCs has the protective effect on the apoptosis of spinal cord cells by reducing the expression of caspase-3 in spinal cord neurons.

Key words: velvet antler polypeptides, glial cell line derived neurotrophic factor, Schwann cells, spinal cord neurons

中图分类号:

Q813.1

张鹏程, 冷向阳, 路博丞, 张远超, 王英, 王旭凯. 鹿茸多肽联合GDNF基因修饰的雪旺细胞对Aβ_25-35诱导脊髓神经元凋亡的保护作用[J]. 吉林大学学报(医学版), 2018, 44(06): 1190-1193.

ZHANG Pengcheng, LENG Xiangyang, LU Bocheng, ZHANG Yuanchao, WANG Ying, WANG Xukai. Protective effect of Schwann cells modified by VAP combined with GDNF gene on apoptosis of spinal cord neurons induced by Aβ_25-35[J]. Journal of Jilin University(Medicine Edition), 2018, 44(06): 1190-1193.

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鹿茸多肽联合GDNF基因修饰的雪旺细胞对Aβ_25-35诱导脊髓神经元凋亡的保护作用

Protective effect of Schwann cells modified by VAP combined with GDNF gene on apoptosis of spinal cord neurons induced by Aβ_25-35

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鹿茸多肽联合GDNF基因修饰的雪旺细胞对Aβ25-35诱导脊髓神经元凋亡的保护作用

Protective effect of Schwann cells modified by VAP combined with GDNF gene on apoptosis of spinal cord neurons induced by Aβ25-35

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鹿茸多肽联合GDNF基因修饰的雪旺细胞对Aβ_25-35诱导脊髓神经元凋亡的保护作用

Protective effect of Schwann cells modified by VAP combined with GDNF gene on apoptosis of spinal cord neurons induced by Aβ_25-35