«上一篇/Previous Article|本期目录/Table of Contents|下一篇/Next Article»

《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:

20卷

期数:

2014年03期

页码:

172-176

栏目:

基础医学

出版日期:

2014-05-20

文章信息/Info

Title:

BMSCs co-transfected with BDNF and TERT improve the ability of cognition and ultra-structure in the hippocampal CA1 region of the rats with vascular dementia

文章编号:

1006-8147(2014)03-0172-05

作者:

王菲¹ ; 常光明² ; 耿鑫³

(1.天津医科大学总医院神经内科,天津300052;2. 天津医科大学总医院检验科,天津300052;3.天津医科大学基础医学院生物化学与分子生物学系;教育部免疫微环境与疾病重点实验室,天津300070)

Author(s):

WANG Fei ¹ ;  CHANG Guang-ming ² ;  GENG Xin ³

（1.Department of Neurology, General Hospital, Tianjin Medical University, Tianjin 300052, China; 2.Department of Clinical Laboratory, General Hospital, Tianjin Medical University, Tianjin 300052, China; 3.Department of Biochemistry and Molecular Biology, Key laboratory of Immune Microenvironment and Diseases of Ministry of Education, Tianjin Medical University, Tianjin 300070, China）

关键词:

血管性痴呆; 骨髓基质干细胞; 脑源性神经营养因子; 端粒酶; 突触素; 大鼠

Keywords:

vascular dementia;  bone marrow mesenchymal stem cells;  brain derived neurotrophic factor;  telomerase;  synaptophysin; rat

分类号:

-

DOI:

-

文献标志码:

A

摘要:

目的：研究脑源性神经营养因子（BDNF）和端粒酶逆转录酶（TERT）联合转染的骨髓间充质干细胞(BMSCs)对血管性痴呆（VD）大鼠学习记忆功能恢复的作用，进一步探讨血管性痴呆的有效治疗途径。方法：经大鼠股骨分离并鉴定BMSCs，构建pLXSN-TERT重组子后转化至BMSCs，软琼脂克隆形成实验检测体外培养的TERT-BMSCs的成瘤性。应用Ad5-BDNF转染TERT-BMSCs，构建BDNF-TERT联合转染的BMSCs。将40只雄性Wistar大鼠随机分为4组：对照组、VD组、BMSCs组、BDNF-TERT-BMSCs组。采用两血管阻断法制备VD模型。采用Morris水迷宫测试方法测试各组大鼠空间学习记忆力。应用RT-PCR和Western blot方法分别检测各组大鼠海马CA1区BDNF、TrkB、SYN基因mRNA和蛋白表达情况，应用透射电镜观察各组大鼠海马CA1区超微结构的变化。结果：行为学实验Morris水迷宫测试结果及超微病理透射电镜观察均显示BMSCs和BDNF-TERT-BMSCs对血管性痴呆大鼠均有改善作用，且BDNF-TERT-BMSCs较BMSCs的作用更为显著。荧光实时定量RT-PCR和Western blot结果显示，BMSCs组和BDNF-TERT-BMSCs组中BDNF、TrkB、SYN mRNA及蛋白表达水平均较痴呆模型组增高（P <0.05），且BDNF-TERT-BMSCs组较BMSCs组增加更为显著，差异有统计学意义（P<0.05）。结论：BDNF-TERT联合转染BMSCs较普通的BMSCs对VD有更好的治疗效果，可明显改善血管性痴呆大鼠的学习记忆功能的恢复。

Abstract:

Objective: To study the effects of BMSCs co-transfected with TERT and BDNF on the expression of BDNF, TrkB and SYN in hippocampal CA1 region of rats model with vascular dementia (VD). Methods: Rat BMSCs were isolated and identified, then transfected by pLXSN-TERT recombinant. Colony formation assay in soft agar was used to detect the tumorigenicity of BMSCs in vitro. TERT-BMSCs were transfected with Ad5-BDNF. Forty male Wistar rats were randomly divided into four groups (10 rats in normal control group, 10 in dementia model group, 10 in BMSCs group, and 10 in BDNF-TERT-BMSCs group). Two-vessel occlusion (2VO) was employed to make VD models. Morris water maze test was done to detect the ability of spatial learning and memory. Then the expression of BDNF, TrkB and SYN in hippocampal CA1 region were determined by real-time RT-PCR and western blot. The ultra-micro structures of synapses in the four groups were detected by transmission electron microscope. Results: The behavior and morphology of the VD rats treated by BMSCs have been enhanced. The average expression of BDNF, TrkB, SYN mRNA and protein in VD group was significantlylower than that in control group (P<0.05). The average expression of BDNF, TrkB, SYN protein in BDNF-TERT-BMSCs group were higher than those in BMSCs group and VD group (P<0.05). The ultra-structure of synapses in BDNF-TERT-BMSCs group was improved significantly compared to BMSCs group and VD group. Conclusion: BMSCs co-transfected with TERT and BDNF could improve the ability of spatial learning and memory in VD rats. The mechanism might be associated with enhancement of the expression of BDNF, TrkB and SYN mRNA and protein and it might have an effect on the synaptic plasticity in hippocampal CA1 area.

参考文献/References:

[1] Ikejima C, Yasuno F, Mizukami K, et al. Prevalence and causes of early-onset dementia in Japan: a population-based study [J]. Stroke, 2009, 40(8):2709 [2] Iadecola C. The pathobiology of vascular dementia [J]. Neuron, 2013, 80(4): 844 [3] Akinyemi R O, Mukaetova-Ladinska E B, Attems J, et al. Vascular risk factors and neurodegeneration in ageing related dementias: Alzheimer’s disease and vascular dementia [J]. Curr Alzheimer Res, 2013, 10 (6): 642 [4] Hu Y, Tan H B, Wang X M, et al.Bone marrow mesenchymal stem cells protect against retinal ganglion cell loss in aged rats with glaucoma [J]. Clin Interv Aging, 2013, 8:1467 [5] Li M D, Atkins H, Bubela T. The global landscape of stem cell clinical trials [J]. Regen Med, 2014, 9(1):27 [6] Van G S, Leerink M, Kakinohana O, et al. Amelioration of motor/sensory dysfunction and spasticity in a rat model of acute lumbar spinal cord injury by human neural stem cell transplantation [J]. Stem Cell Res Ther, 2013, 28, 4(3):57 [7] Banfi A, Bianchi G, Notaro R, et al. Replicative aging and gene expression in long-term cultures of human bone marrow stromal cells [J]. Tissue Eng, 2002, 8 (6):901 [8] Baxter M A, Wynn R F, Jowitt S N, et al. Study of telomere length reveals rapid aging of human marrow stromal cells following in vitro expansion [J]. Stem Cells, 2004, 22 (5): 675 [9] Bianchi G, Banfi A, Mastrogiacomo M, et al. Ex vivo enrichment of mesenchymal cell progenitors by fibroblast growth factor 2 [J]. Exp Cell Res, 2003, 287 (1): 98 [10] Chen Z Z, Xu R X, Jiang X D, et al. Lack of telomerase activity in rabbit bone marrow stromal cells during differentiation along neural pathway [J]. Chin J Traumatol, 2006, 9 (4): 201 [11] Bokara K K, Kwon K H, Nho Y, et al. Retroviral expression of arginine decarboxylase attenuates oxidative burden in mouse cortical neural stem cells [J]. Stem Cells Dev, 2011, 20 (3):527 [12] Ni J, Ohta H, Matsumoto K,et al. Progressive cognitive impairment following chronic cerebral hypoperfusion induced by permanent occlusion of bilateral carotid arteries in rats [J]. Brain Res, 1994, 653(1-2):231 [13] Wang F, Geng X, Tao H Y, et al. The restoration after repetitive transcranial magnetic stimulation treatment on cognitive ability of vascular dementia rats and its impacts on synaptic plasticity in hippocampal CA1 area [J]. J Mol Neurosci, 2010, 41(1):145 [14] Boccardi V, Herbig U. Telomerase gene therapy: a novel approach to combat aging [J]. EMBO Mol Med, 2012, 4(8):685 [15] Razavi S, Razavi M R, Kheirollahi-Kouhestani M, et al. Co-culture with neurotrophic factor secreting cells induced from adipose-derived stem cells: Promotes neurogenic differentiation [J]. Biochem Biophys Res Commun, 2013, 440(3):381 [16] Tang H, Xiang Y, Jiang X, et al. Dual expression of hTERT and VEGF prolongs life span and enhances angiogenic ability of aged BMSCs [J]. Biochem Biophys Res Commun, 2013, 440(4):502 [17] Cheeran B, Talelli P, Mori F, et al. A common polymorphism in the brain derived neurotrophic factor gene (BDNF) modulates human cortical plasticity and the response to rTMS [J]. J Physiol, 2008, 586 (23):5717 [18] Sun S, Chen G, Xu M, et al. Differentiation and Migration of Bone Marrow Mesenchymal Stem Cells Transplanted through the Spleen in Rats with Portal Hypertension [J]. PLoS One, 2013, 8(12):e83523

相似文献/References:

备注/Memo

备注/Memo:

基金项目 国家自然科学基金资助项目（81100805）；天津市高校科技发展计划基金资助项目（20080107）；基因工程教育部重点实验室开放研究基金资助项目（201302）
作者简介 王菲（1979-），女，主治医师，博士，研究方向：分子神经病学；通信作者：耿鑫，E-mail: gengx@tmu.edu.cn。

常用功能

更新日期/Last Update: 2014-05-12