Nurr1基因修饰胚胎中脑神经干细胞移植治疗帕金森病

乔奕胜; 陈孝祥; 徐蛟天; 王威; 张超; 宋晓斌; 杨智勇; 邓兴力

doi:10.12259/j.issn.2095-610X.S20210801

Nurr1基因修饰胚胎中脑神经干细胞移植治疗帕金森病

doi: 10.12259/j.issn.2095-610X.S20210801

乔奕胜^{1, 2},
陈孝祥¹,
徐蛟天¹,
王威¹,
张超¹,
宋晓斌¹,
杨智勇¹,
邓兴力^1, ,

1.
昆明医科大学第一附属医院神经外一科，云南昆明　650032
2.
云南省滇东北区域中心医院神经医学中心，云南昭通　547000

基金项目: 国家自然科学基金资助项目（81360197）；云南省科技厅-昆明医科大学应用基础研究联合专项基金资助项目（2017FB130）

详细信息

作者简介:
乔奕胜(1993～)，男，云南安宁人，医学硕士，主要从事神经外科临床工作

通讯作者:
邓兴力，E-mail：dxlkmmu@163.com

中图分类号: R742.5
计量
- 文章访问数: 4299
- HTML全文浏览量: 2262
- PDF下载量: 41
- 被引次数: 0
出版历程
- 收稿日期: 2021-06-13
- 网络出版日期: 2021-08-04
- 刊出日期: 2021-08-04

Transplantation of Nurr1 Gene-modified Embryo Midbrain Neural Stem Cells for Treatment in Parkinson’ s Disease

1.
Dept. of Neurosurgery，The 1st Affiliated Hospital, Kunming Medical University， Kunming Yunnan 650032
2.
Neurology Center of Northeast Yunnan Regional Central Hospital，Zhaotong Yunnan 547000，China

摘要

摘要: 目的探讨移植Nurr1基因修饰胚胎中脑神经干细胞对帕金森大鼠的治疗效果。方法（1）建立SD大鼠帕金森病模型；（2）原代培养SD大鼠胚胎中脑神经干细胞，并于神经干细胞过表达Nurr1；（3）移植治疗分组：假手术组（A组），移植神经干细胞组（B组），移植Nurr1修饰神经干细胞组（C组）；（4）在移植后3周、6周、9周和12周分别对各组大鼠进行行为学测试，观察各组症状改善情况；（5）移植12周后，分别检测各组中Nurr1、TH蛋白表达情况和Nurr1荧光情况。结果（1）原代培养大鼠胚胎中脑神经干细胞，并于神经干细胞成功过表达Nurr1基因。（2）免疫荧光、Western Blot证实过表达Nurr1可以促进神经干细胞向多巴胺能神经元转化（P < 0.05）。（3）移植Nurr1修饰神经干细胞可以有效的改善帕金森大鼠的症状（P < 0.05），Western Blot证实小胶质细胞可显著促进移植后的神经干细胞在分化为多巴胺能神经元（P < 0.05）。（4）移植后Nurr1能促进神经干细胞向多巴胺能神经元转化，提高移植神经干细胞的存活，达到更好治疗效果。结论 Nurr1基因可以促进神经干细胞向多巴胺能神经元转化；移植修饰Nurr1神经干细胞可以有效治疗帕金森大鼠。
- Nurr1 /
- 胚胎中脑神经干细胞 /
- 帕金森病
Abstract: Objective To investigate the therapeutic effect of transplanted Nurr1 gene-modified embryo midbrain neural stem cells in Parkinson’ s rats. Methods 1. Establishment of Parkinson’ s disease animal model in SD rats. 2. Primary culture of embryo midbrain neural stem cells （NCSs） and Nurr1 gene-modified neural stem cells. 3. Grouping: sham operation group （sham group）, Transplanted neural stem cell group （NCS group）, transplanted Nurr1 gene-modified neural stem cells group （NNCS group）. 4. At 3 weeks, 6 weeks, 9 weeks, and 12 weeks after the transplantation, the behavioral tests of rats in each group were performed to observe the improvement of symptoms in each group. 5. After 12 weeks of transplantation, the expression of Nurr1, TH protein and Nurr1 fluorescence were detected in each group. Results 1. Primary cultured neural stem cells, and successfully overexpressed Nurr1 in neural stem cells. 2. Overexpression of Nurr1 could promote the transformation of neural stem cells into dopaminergic neurons （P < 0.05）, which was confirmed by immunofluorescence and Western Blot. 3. Transplantation of NNCS can effectively improve the symptoms of rats with Parkinson’ s （P < 0.05）. 4. After transplantation, Nurr1 can promote the transformation of neural stem cells into dopaminergic neurons, greatly improve the survival of transplanted neural stem cells and achieve better therapeutic effect. Conclusion Nurr1 can promote the transformation of neural stem cells into dopaminergic neurons. Transplantation of overexpression of Nurr1 neural stem cells can effectively treat Parkinson’ s rats.
- Nurr1 /
- Embryonic stem cells /
- Parkinson disease

HTML全文

图 1 神经干细胞的培养及鉴定

A：神经干细胞原代培养7 d；B、C：巢蛋白（Nestin）鉴定阳性。

Figure 1. Culture and identification of neural stem cells

下载: 全尺寸图片幻灯片

图 2 过表达Nurr1和对神经干细胞分化的影响

A：转染72 h后荧光显微镜检测Nurr1呈绿色；B、C：转染7 d后RT-PCR检测Nurr1的mRNA表达；D：分化培养7 d后检测TH荧光呈红色；E、F、G：分化7 d后Western检测Nurr1和TH表达。　　　　　　　　　　　　　　　　　　　　　与对照组比较，^*** P < 0.001。

Figure 2. Overexpression of Nurr1 and its effect on differentiation of neural stem cells

下载: 全尺寸图片幻灯片

图 3 细胞移植后分析阿扑吗啡诱导的旋转评分

各组分别在移植后3、6、9、12周，利用阿扑吗啡诱导检测转圈行为　　　　　　　　　　　　与C组比较，^*P < 0.05， ^**P < 0.001。

Figure 3. Apomorphine-induced rotation scores after cell transplantation.

下载: 全尺寸图片幻灯片

图 4 移植Nurr1后神经干细胞向多巴胺能神经元转化的情况

A：各组Nurr1和TH蛋白的表达水平；B、C：C组与对照组及B相比，Nurr1和TH表达上调；D、E：移植治疗12周后。　　　　　　　　　　　　　　　　　　　与NNCS组比较，^* P < 0.05，^**P < 0.01。

Figure 4. Transformation of Neural Stem Cells into dopaminergic neurons after Nurr1 transplantation

下载: 全尺寸图片幻灯片

参考文献(18)

[1]	Balestrino R,Schapira A H V. Parkinson disease[J]. Eur J Neurol,2020,27(1):27-42.
[2]	陈彪. 左旋多巴在帕金森病治疗中的地位及进展[J]. 中国临床神经科学,2017,25(5):546-550. doi: 10.3969/j.issn.1008-0678.2017.05.012
[3]	Zhu B,Caldwell M,Song B. Development of stem cell-based therapies for Parkinson’ s disease[J]. Int J Neurosci,2016,126(11):955-962.
[4]	Zhang Q,Chen W,Tan S,et al. Stem cells for modeling and therapy of parkinson’ s disease[J]. Hum Gene Ther,2017,28(1):85-98. doi: 10.1089/hum.2016.116
[5]	刘罡,吴毅,贾杰,等. 神经干细胞体外分化抗原表达的研究[J]. 中国运动医学杂志,2009,28(1):55-59.
[6]	Li K, Li J, Zheng J, Qin S. Reactive astrocytes in neurodegenerative diseases[J]. Aging Dis. 2019; 10(3): 664-675.
[7]	Barker R A,Drouin Ooellet J,Parmar M. Cell-based therapies for Parkinson disease-past insights and future potential[J]. Nat Rev Neurol,2015,11(9):492-503. doi: 10.1038/nrneurol.2015.123
[8]	徐蛟天,陈孝祥,王威,等. Nurr1对小胶质细胞联合神经干细胞共培养促进神经干细胞向多巴胺神经元分化作用研究[J]. 国际神经病学神经外科学杂志,2018,45(01):52-57.
[9]	Liu W,Gao Y,Chang N. Nurr1 overexpression exerts neuroprotective and anti-inflammatory roles via down-regulating CCL2 expression in both in vivo and in vitro Parkinson’ s disease models[J]. Biochem Biophys Res Commun,2017,482(4):1312-1319. doi: 10.1016/j.bbrc.2016.12.034
[10]	Kadkhodaei B,Ito T,Joodmardi E,et al. Nurr1 is required for maintenance of maturing and adult midbrain dopamine neurons[J]. J Neurosci,2009,29(50):15923-15932. doi: 10.1523/JNEUROSCI.3910-09.2009
[11]	Radad K,Moldzio R,AL-Shraim M,et al. Recent advances on the role of neurogenesis in the adult brain:Therapeutic potential in parkinson’ s and Alzheimer’ s diseases[J]. CNS Neurol Disord Drug Targets,2017,16(7):740-748.
[12]	O’ sullivan S S,Johnson M,Williams D R,et al. The effect of drug treatment on neurogenesis in parkinson's disease[J]. Mov Disord,2011,26(1):45-50. doi: 10.1002/mds.23340
[13]	Madhavan L,Daley B F,Paumier K L,et al. Transplantation of subventricular zone neural precursors induces an endogenous precursor cell response in a rat model of Parkinson’ s disease[J]. J Comp Neurol,2009,515(1):102-115. doi: 10.1002/cne.22033
[14]	Dong J,Liu X,Wang Y,et al. Nurr1(Cd11bcre) conditional knockout mice display inflammatory injury to nigrostriatal dopaminergic neurons[J]. Glia,2020,68(10):2057-2069. doi: 10.1002/glia.23826
[15]	Wang X,Zhuang W,Fu W,et al. The lentiviral-mediated Nurr1 genetic engineering mesenchymal stem cells protect dopaminergic neurons in a rat model of Parkinson’ s disease[J]. Am J Transl Res,2018,10(6):1583-1599.
[16]	Smith G A,Rocha E M,Rooney T,et al. A Nurr1 agonist causes neuroprotection in a Parkinson’ s disease lesion model primed with the toll-like receptor 3 dsRNA inflammatory stimulant poly (I:C)[J]. PLoS One,2015,10(3):e0121072. doi: 10.1371/journal.pone.0121072
[17]	Ahmed A,Isaksen T J,Yamashita T. Protocol for mouse adult neural stem cell isolation and culture[J]. STAR Protoc,2021,2(2):100522. doi: 10.1016/j.xpro.2021.100522
[18]	Raina A,Mahajani S,Bahr M,et al. Neuronal trans-differentiation by traNCSription factors ascl1 and Nurr1:Induction of a dopaminergic neurotransmitter phenotype in cortical GABAergic neurons[J]. Mol Neurobiol,2020,57(1):249-260. doi: 10.1007/s12035-019-01701-x

相关文章(20)

[1]	罗丽, 孙月, 王禹雪, 陆永萍. 实时剪切波弹性成像评估帕金森病患者股四头肌肌肉及肌腱肌张力的变化, 昆明医科大学学报.
[2]	蔺小英, 谭艳, 杨磊. 双重任务训练对帕金森病患者运动功能的影响, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20230110
[3]	杨梦姝, 孔晓云, 李欢, 邹延威, 王义豪, 董慈. 失眠的认知行为疗法对帕金森病伴失眠患者影响的临床研究, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20230806
[4]	李波, 角建林, 张传意, 邱娅琼, 国灏, 孙怀昌, 俞志成. MPTP诱导树鼩帕金森病模型的脑电图描记与分析, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20210506
[5]	乔奕胜, 陈孝祥, 徐蛟天, 王威, 张超, 宋晓斌, 杨智勇, 邓兴力. Nurr1基因修饰胚胎中脑神经干细胞移植治疗帕金森病, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20210919
[6]	周舒, 陈武龙, 徐君铭, 王淑芬. 内源性神经干细胞治疗多发性硬化的研究进展, 昆明医科大学学报.
[7]	张凤兰, 赵瑜, 袁婧, 杨璐军, 鲁琳. 人参皂苷Rg3对体外培养小鼠神经干细胞增殖的影响, 昆明医科大学学报.
[8]	李余斌, 耿鑫, 戚仁莉, 李经辉, 蒋红高, 尹雷, 柳安雄, 李雨珊, 唐佳, 余化霖. 双侧STN-DBS治疗中晚期帕金森患者运动症状短期疗效, 昆明医科大学学报.
[9]	赵瑜, 袁婧, 刘建军, 鲁琳. 小鼠胚胎脑源与脊髓源神经干细胞增殖能力的比较, 昆明医科大学学报.
[10]	黄微, 戴萍, 殷燕, 张玲, 贾帆, 郭玲, 李劲涛. 帕金森病冻结步态与躯体形式障碍临床分析, 昆明医科大学学报.
[11]	施国林. 脑深部电刺激术治疗帕金森病40例报道, 昆明医科大学学报.
[12]	鲍娟. 帕金森病患者轻度认知损害与胰岛素抵抗的相关性, 昆明医科大学学报.
[13]	和茵. 脑电深部刺激法治疗帕金森病的护理体会, 昆明医科大学学报.
[14]	黄微. 原发性震颤误诊为帕金森病25例临床分析, 昆明医科大学学报.
[15]	黄云燕. 多氯联苯对胎鼠神经干细胞Nnat基因表达的影响, 昆明医科大学学报.
[16]	王波. 3.0T MR磁敏感加权成像对早期帕金森病黑质致密带的研究, 昆明医科大学学报.
[17]	耿鑫. 脑深部电刺激治疗帕金森病1例报道, 昆明医科大学学报.
[18]	. 体外培养神经干细胞CNTF、TGF-β1和IGF-1的表达, 昆明医科大学学报.
[19]	丁鹏. 新生大鼠海马神经干细胞表达趋化因子受体CCR2的体外研究, 昆明医科大学学报.
[20]	杨金伟. 神经干细胞与嗅鞘细胞移植对脊髓全横断大鼠后肢运动功能的影响, 昆明医科大学学报.