NT-3-HUMSCs联合基因沉默SOCS3治疗SD大鼠脊髓损伤后的运动功能分析

白刚; 张洪钿; 赖军; 罗林; 左频; 范耀东

NT-3-HUMSCs联合基因沉默SOCS3治疗SD大鼠脊髓损伤后的运动功能分析

1. 昆明医科大学第三附属医院
2. 中国人民解放军陆军总医院
3. 重庆市人民医院

基金项目:

基金：云南省教育厅科学研究基金重点科研基金资助项目 (2014Z054);

计量
- 文章访问数: 1917
- HTML全文浏览量: 682
- PDF下载量: 54
- 被引次数: 0
出版历程
- 收稿日期: 2017-09-21

An Analysis of Movement Function of SD Rats' Spinal Cord Injury After Joint Therapy Using NT-3-HUMSCs and SOCS3 Gene Silencing

Funds:

基金：云南省教育厅科学研究基金重点科研基金资助项目 (2014Z054);

摘要

摘要: 目的 NT-3-HUMSCs联合基因沉默SOCS3治疗SD大鼠脊髓损伤, 以期改善损伤神经运动功能.方法 (1) 用贴壁法体外培养人脐带间充质细胞 (HUMSC) , 同时进行分离, 提纯和鉴定; (2) 构建NT-3基因真核表达载体, 利用基因转染技术将其转入HUMSC, 构建NT-3-HUMSC细胞, 体外检测其存活情况及NT-3表达情况; (3) 筛选作用于SOCS3的特异性靶点, 进行序列同源性分析, 设立阴性对照, 设计并合成SOCS3-siRNA, 同时在体外检测功能; (4) 建立SD大鼠脊髓损伤模型分为为:假手术组10只;T12全脊髓横断损伤模型40只, 随机分为4组, 生理盐水治疗组10只;siRNA+NT-3-HUMSCs治疗组10只;NT-3-HUMSCs治疗组10只;SOCS3-siRNA治疗组10只.以上各组造模成功后, 分别存活1、2月和3月进行运动功能评价.结果 (1) siRNA+NT-3-HUMSCs联合治疗SD大鼠脊髓损伤组BBB评分明显高于NT-3-HUMSCs、siRNA和生理盐水组, 差异有统计学意义 (P<0.05) ; (2) siRNA+NT-3-HUMSCs联合治疗SD大鼠脊髓损伤组爬网格实验表明神经功能恢复明显高于NT-3-HUMSCs、siRNA和生理盐水组, 差异有统计学意义 (P<0.05) .结论 NT-3-HUMSCs联合基因沉默SOCS3治疗SD大鼠脊髓损伤, 可以改善SD大鼠脊髓损伤的运动功能.
- 脊髓损伤 /
- SOCS3 /
- siRNA /
- NT-3基因 /
- 基因转染
Abstract: Objective To achieve the purpose of promoting movement function of the injury nerve by using the joint therapy of NT-3-HUMSCs and SOCS3 gene silencing on SD rats' spinal cord injury. Methods (1) We used adherence method in vitro human umbilical cord-derived mesenchymal cells ( HUMSC) during separation, purification and identification. (2) Then constructed NT-3 gene eukaryotic expression vector, which was transfected into its HUMSC, and constructed NT-3-HUMSC cell survival in vitro assay conditions and NT-3 expression. (3) We selected specific targets for SOCS3 screening and for sequence homology analysis.A negative control group was established. siRNA was designed and synthesized in vitro detection. (4) SD rats with spinal cord injury model were divided into two categories: (1) sham group with 10 rats; (2) T12 whole spinal cord injury model with 40 rats. The 40 rats were randomly divided into four groups with 10 rats in each group (saline treatment group, siRNA + NT-3-HUMSCs treatment group, NT-3-HUMSCs treatment group and siRNA treated group) . Motor function of the rats were evaluated respectively in 1, 2 and 3 months after the modeling was established successfully.Results (1) siRNA + NT-3-HUMSCs treatment group's BBB scores was significantly higher than NT-3-HUMSCs, SOCS3-siRNA and physiological saline groups (P <0.05) . (2) The grid climbing experiments showed that the neural functional recovery performed better in siRNA + the NT-3-HUMSCs treatment group compared to the NT-3-HUMSCs, SOCS3-siRNA and physiological saline groups (P <0.05) . Conclusion The NT-3-HUMSCs joint SOCS3 gene silencing in the treatment of SD rat spinal cord injury can improve the motor function of SD rat spinal cord injury.
- Spinal cord injury /
- SOCS3 /
- siRNA /
- NT-3 gene /
- Gene transfection

HTML全文

参考文献(18)

[1]	[1]WALTHERS C M, SEIDLITS S K.Gene delivery strategies to promote spinal cord repair[J].Biomarker Insights, 2015, 10 (Suppl 1) :11.
[2]	[2]HAGEN E M, REKAND T, GILHUS N E, et al.Traumatic spinal cord injuries-incidence, mechanisms and course[J].Tidsskrift for Den Norske Laegeforening:Tidsskrift for Praktisk Medicin, Ny Raekke, 2012, 132 (7) :831-837.
[3]	[3]RAMER L M, RAMER M S, BRADBURY E J.Restoring function after spinal cord injury:towards clinical translation of experimental strategies[J].The Lancet Neurology, 2014, 13 (12) :1241-1256.
[4]	[4]RAMONY, CAJAL, S.Degenerationandregenerationofthe nervous system[M].New York:Oxford University Press, 1928:557.
[5]	[5]SCHIRA J, GASIS M, ESTRADA V, et al.Significant clinical, neuropathological and behavioural recovery from acute spinal cord trauma by transplantation of a well-defined somatic stem cell from human umbilical cord blood[J].Brain, 2012, 135 (2) :431-446.
[6]	[6]CAO Q, WHITTEMORE S R.Cell transplantation:stem cells and precursor cells[J].Handbook of Clinical Neurology, 2011, 109:551-561.
[7]	[7]DAVIESSJA, SHIHCH, NOBLEM, etal.Transplantationof specifichumanastrocytespromotesfunctionalrecovery after spinal cord injury[J].Plo S One, 2011, 6 (3) :e17328.
[8]	[8]GUESTJ, SANTAMARIAAJ, BENAVIDESFD.Clinical translation ofautologous Schwann cell transplantation for the treatment of spinal cord injury[J].Current Opinion in Organ Transplantation, 2013, 18 (6) :682.
[9]	[9]NAKAMURAM, OKANOH.Celltransplantationtherapies for spinal cord injury focusing on induced pluripotent stem cells[J].Cell Research, 2012, 23 (1) :70-80.
[10]	[10]KRABBEC, ZIMMERJ, MEYERM.Neuraltransdifferentiationofmesenchymalstemcellsa critical review[J].Apmis, 2005, 113 (11-12) :831-844.
[11]	[11]PHINNEY D G, ISAKOVA I.Plasticity and therapeutic potential of mesenchymal stem cells in the nervous system[J].Current Pharmaceutical Design, 2005, 11 (10) :1255-1265.
[12]	[12]CHEN Y, TENG F Y H, TANG B L.Coaxing bone marrow stromalmesenchymalstemcellstowardsneuronal differentiation:progress and uncertainties[J].Cellular and Molecular Life Sciences CMLS, 2006, 63 (14) :1649-1657.
[13]	[13]BARZILAY R, MELAMED E, OFFEN D.Introducing transcription factors to multipotent mesenchymal stem cells:making transdifferentiation possible[J].Stem Cells, 2009, 27 (10) :2509-2515.
[14]	[14]KOSHIZUKA S, OKADA S, OKAWA A, et al.Transplanted hematopoietic stem cells from bone marrow differentiate into neural lineage cells and promote functional recovery after spinal cord injury in mice[J].Journal of Neuropathology&Experimental Neurology, 2004, 63 (1) :64-72.
[15]	[15]YANGCC, SHIHYH, KOMH, etal.Transplantationofhuman umbilical mesenchymal stem cells from Wharton's jelly aftercompletetransectionoftheratspinalcord[J].PLo SOne, 2008, 3 (10) :e3336.
[16]	[16]ZHANG L, ZHANG H T, HONG S Q, et al.Cografted Wharton’s jelly cells-derived neurospheres and BDNF promote functional recovery after rat spinal cord transection[J].Neurochemical Research, 2009, 34 (11) :2030-2039.
[17]	[17]HU S L, LUO H S, LI J T, et al.Functional recovery in acute traumatic spinal cord injury after transplantation of human umbilical cord mesenchymal stem cells[J].Critical Care Medicine, 2010, 38 (11) :2181-2189.
[18]	[18]SHANG A J, HONG S Q, XU Q, et al.NT-3-secreting human umbilical cord mesenchymal stromal cell transplantation for the treatment of acute spinal cord injury in rats[J].Brain Research, 2011, 1391:102-113.

相关文章(20)

[1]	周磊, 吕雯雯, 段永忠, 钱雯, 程继帅. 靶向抗HSV-1的siRNA研究进展, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20240901
[2]	吴疆, 马和, 江金鋆, 刘娜, 赵瑞欢, 何越峰. 砒霜厂工人砷暴露与EGFR、PTEN、Kras、PIK3CA四个基因DNA损伤的关系, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20230717
[3]	谢环, 田洁, 黄娅芬. shRNA Twist基因对人绒毛膜癌细胞JEG-3增殖、侵袭和凋亡的影响, 昆明医科大学学报.
[4]	王福科, 张红, 杨桂然, 肖渝, 何川, 宋恩, 李彦林. 携载增强型绿色荧光蛋白基因慢病毒转染骨髓间充质干细胞示踪方法, 昆明医科大学学报.
[5]	李勤学, 李传静, 王锋. 慢病毒介导siRNA沉默MMP-3对大鼠创伤性骨性关节炎模型软骨退变的影响, 昆明医科大学学报.
[6]	陈珊珊, 易敏春, 周国忠, 普跃昌, 胡毅, 韩米华, 金华. CNTF干扰对BMSCs动脉移植缺血再灌注损伤脊髓下游信号通路STAT3/Caspase-9的影响, 昆明医科大学学报.
[7]	白刚, 张洪钿, 赖军, 罗林, 左频, 范耀东. NT-3-HUMSCs联合基因沉默SOCS3治疗SD大鼠脊髓损伤后的神经再生修复, 昆明医科大学学报.
[8]	王以婷, 杨卫红, 赵云龙, 曾昭书, 张莉蓉. CYP3A4内含子10对CYP3A4基因表达的增强子作用, 昆明医科大学学报.
[9]	周琦. 催眠治疗对脊髓损伤患者康复的影响, 昆明医科大学学报.
[10]	单可记. 氨茶碱对急性颈髓损伤并心动过缓患者近期疗效探讨, 昆明医科大学学报.
[11]	邵文萍. 云南宣威肺腺癌细胞XWLC-05转染HLA-A*0201的抗肿瘤研究, 昆明医科大学学报.
[12]	陈绍春. 用慢病毒转染技术实现Gnaq基因在SH-SY5Y细胞中持续稳定高表, 昆明医科大学学报.
[13]	刘为军. 慢病毒载体介导CD1d和GFP融合基因转染PANC-1细胞的实验研究, 昆明医科大学学报.
[14]	袁聪. 甲泼尼龙对受损脊髓组织中抗菌肽相关基因表达的调节, 昆明医科大学学报.
[15]	. 大鼠变应性鼻炎模型中T-bet/GATA-3基因表达的研究, 昆明医科大学学报.
[16]	. 肺腺癌A549细胞株GDNF、BDNF、NT3和NT4的表达, 昆明医科大学学报.
[17]	李昆仑. RNAi抑制Survivin基因的表达对乳腺癌SKBr-3细胞的影响, 昆明医科大学学报.
[18]	李玉. 体外培养神经干细胞NGF、BDNF和NT-3及其受体trkA、trkB和trkC 的表达, 昆明医科大学学报.
[19]	段艳萍. GAP-43治疗大鼠完全性脊髓损伤后Nogo-A在神经元的表达, 昆明医科大学学报.
[20]	朱兴宝. ORMA40载体介导生存素基因转染嗅神经鞘细胞, 昆明医科大学学报.