山东医药
山東醫藥
산동의약
SHANDONG MEDICAL JOURNAL
2014年
13期
1-3
,共3页
痴呆,血管性%端粒酶%骨髓基质干细胞%突触可塑性%大鼠
癡呆,血管性%耑粒酶%骨髓基質榦細胞%突觸可塑性%大鼠
치태,혈관성%단립매%골수기질간세포%돌촉가소성%대서
dementia,vascular%telomerase%bone marrow mesenchymal stem cells%synaptic plasticity%rats
目的:研究端粒酶逆转录酶(TERT)基因转染的骨髓基质干细胞(BMSCs)对血管性痴呆(VD)大鼠学习记忆功能的恢复及其对海马CA1区突触可塑性影响的分子机制。方法经大鼠股骨及胫骨分离并鉴定BMSCs,构建反转录病毒pLXSN-TERT重组子并转染BMSCs,软琼脂克隆形成实验检测体外培养的TERT-BMSCs的成瘤性。将36只雄性Wistar大鼠随机分为正常对照组、痴呆模型组、BMSCs组和TERT-BMSCs组。采用双侧颈总动脉永久性结扎方法制作VD大鼠模型,Morris水迷宫测试法检测各组学习记忆能力,透射电镜观察海马CA1区超微结构变化;免疫组织化学方法观察海马CA1区BDNF、NMDAR1、SYN蛋白表达。结果 Morris水迷宫测试及透射电镜观察均显示,BMSCs组、TERT-BMSCs组较痴呆模型组学习记忆能力、海马CA1区超微结构明显改善(P均<0.05),且TERT-BMSCs组比BMSCs组改善更为明显(P均<0.05)。 BMSCs组、TERT-BMSCs组BDNF、NMDAR1、SYN蛋白阳性细胞着色的平均灰度值比痴呆模型组增高(P均<0.05),且TERT-BMSCs组比BMSCs组增高更为显著(P均<0.05)。结论 TERT基因转染的BMSCs比普通BM-SCs对VD的康复治疗作用更为显著,其分子机制可能与增加大鼠海马CA1区BDNF、NMDAR1和SYN的表达,影响了突触可塑性有关。
目的:研究耑粒酶逆轉錄酶(TERT)基因轉染的骨髓基質榦細胞(BMSCs)對血管性癡呆(VD)大鼠學習記憶功能的恢複及其對海馬CA1區突觸可塑性影響的分子機製。方法經大鼠股骨及脛骨分離併鑒定BMSCs,構建反轉錄病毒pLXSN-TERT重組子併轉染BMSCs,軟瓊脂剋隆形成實驗檢測體外培養的TERT-BMSCs的成瘤性。將36隻雄性Wistar大鼠隨機分為正常對照組、癡呆模型組、BMSCs組和TERT-BMSCs組。採用雙側頸總動脈永久性結扎方法製作VD大鼠模型,Morris水迷宮測試法檢測各組學習記憶能力,透射電鏡觀察海馬CA1區超微結構變化;免疫組織化學方法觀察海馬CA1區BDNF、NMDAR1、SYN蛋白錶達。結果 Morris水迷宮測試及透射電鏡觀察均顯示,BMSCs組、TERT-BMSCs組較癡呆模型組學習記憶能力、海馬CA1區超微結構明顯改善(P均<0.05),且TERT-BMSCs組比BMSCs組改善更為明顯(P均<0.05)。 BMSCs組、TERT-BMSCs組BDNF、NMDAR1、SYN蛋白暘性細胞著色的平均灰度值比癡呆模型組增高(P均<0.05),且TERT-BMSCs組比BMSCs組增高更為顯著(P均<0.05)。結論 TERT基因轉染的BMSCs比普通BM-SCs對VD的康複治療作用更為顯著,其分子機製可能與增加大鼠海馬CA1區BDNF、NMDAR1和SYN的錶達,影響瞭突觸可塑性有關。
목적:연구단립매역전록매(TERT)기인전염적골수기질간세포(BMSCs)대혈관성치태(VD)대서학습기억공능적회복급기대해마CA1구돌촉가소성영향적분자궤제。방법경대서고골급경골분리병감정BMSCs,구건반전록병독pLXSN-TERT중조자병전염BMSCs,연경지극륭형성실험검측체외배양적TERT-BMSCs적성류성。장36지웅성Wistar대서수궤분위정상대조조、치태모형조、BMSCs조화TERT-BMSCs조。채용쌍측경총동맥영구성결찰방법제작VD대서모형,Morris수미궁측시법검측각조학습기억능력,투사전경관찰해마CA1구초미결구변화;면역조직화학방법관찰해마CA1구BDNF、NMDAR1、SYN단백표체。결과 Morris수미궁측시급투사전경관찰균현시,BMSCs조、TERT-BMSCs조교치태모형조학습기억능력、해마CA1구초미결구명현개선(P균<0.05),차TERT-BMSCs조비BMSCs조개선경위명현(P균<0.05)。 BMSCs조、TERT-BMSCs조BDNF、NMDAR1、SYN단백양성세포착색적평균회도치비치태모형조증고(P균<0.05),차TERT-BMSCs조비BMSCs조증고경위현저(P균<0.05)。결론 TERT기인전염적BMSCs비보통BM-SCs대VD적강복치료작용경위현저,기분자궤제가능여증가대서해마CA1구BDNF、NMDAR1화SYN적표체,영향료돌촉가소성유관。
Objective To study the molecular mechanisms underlying the effects of bone marrow mesenchymal stem cells (BMSCs) transfected with telomerase reverse transcriptase (TERT) on the restoration of learning and memory ability and the impact on synaptic plasticity in the hippocampal CA 1 area of rats 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 TERT-BMSCs in vitro .Thirty-six male Wistar rats were randomly divided into the con-trol group, VD group, BMSCs group and TERT-BMSCs group.Two-vessel occlusion (2VO) was employed to make VD models .Morris water maze test was done for the detection of spatial learning and memory ability .Then the ultrastructures of synapses in the four groups were detected by transmission electron microscope .The expression of BDNF , NMDAR1 and SYN in hippocampal CA 1 region was determined by immunohistochemistry .Results Both Morris water maze test and transmission electron microscope showed that the behavior and morphology were improved in the BMSCs and TERT -BMSCs groups as compared with that of the VD group (all P<0.05), and the TERT-BMSCs group was better (all P<0.05). The average expression levels of BDNF , NMDAR1 and SYN protein in the BMSCs and TERT-BMSCs groups were higher than those in the VD group (all P<0.05), and TERT-BMSCs group had more significant effects than BMSCs group (all P<0.05).Conclusions TERT-BMSCs can more significantly improve the ability of spatial learning and memory in VD rats than regular BMSCs .This mechanism might be associated with enhancement of the expression of BDNF , NMDAR1 and SYN protein in hippocampal CA 1 region and affecting the synaptic plasticity .