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| 小鼠Calbindin-D28k基因体外表达及其抗凋亡作用的研究 | |
| 引用本文： | 朱孝荣,朱园园,李富珍,袁红花,孙怀昌,高殿帅.小鼠Calbindin-D28k基因体外表达及其抗凋亡作用的研究[J].神经解剖学杂志,2010,26(1). |
| 作者姓名： | 朱孝荣 朱园园 李富珍 袁红花 孙怀昌 高殿帅 |
| 作者单位： | 1. 扬州大学兽医学院预防兽医教研室,扬州,215009;徐州医学院神经生物学教研室,徐州,221002 2. 徐州医学院神经生物学教研室,徐州,221002 3. 扬州大学兽医学院预防兽医教研室,扬州,215009 |
| 基金项目： | 国家自然科学基金，江苏省高校自然科学基础研究面上项目 |
| 摘 要： | 目的:研究钙结合蛋白D28k(calbindin-D28k,CaBP)体外表达及其对神经元细胞抗凋亡作用。方法:RT-PCR从小鼠脑组织中扩增出小鼠CaBP的cDNA片段;重组构建pTH-CB质(pcDNA3-TH-CB),脂质体包裹pTH-CB质粒转染MN-9D细胞(多巴胺能杂交瘤神经元细胞);RT-PCR、免疫沉淀法检测CaBP的cDNA在细胞内表达情况;Hoechst染色、Caspase-3活性检测、AnnexinV/PI法对CaBP过表达的抗凋亡作用进行检测。结果:小鼠脑组织RT-PCR扩增出785bp条带,测序结果与Genebank中报道完全一致。脂质体包裹重组构建pTH-CB质粒转染MN-9D细胞后,转染pTH-CB质粒的MN-9D细胞中,CaBP的mRNA表达量和CaBP水平显著高于转染pc-DAN3组和正常MN-9D细胞组(P0.05),3种细胞凋亡检测方法结果显示,CaBP过表达的细胞抗凋亡能力显著提高。结论:体外克隆CaBP的cDNA能在细胞内表达,过表达的CaBP通过抑制细胞的caspase-3活性,起抗凋亡作用。 |
| 关 键 词： | 钙结合蛋白D28k 基因表达 凋亡 小鼠 |
In vitro expression and anti-apoptotic effect of mouse calbindin-D28k |
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| Zhu Xiaorong,Zhu Yuanyuan,Li Fuzhen,Yuan Honghua,Sun Luaichang,Gao Dianshuai.In vitro expression and anti-apoptotic effect of mouse calbindin-D28k[J].Chinese Journal of Neuroanatomy,2010,26(1). | |
| Authors: | Zhu Xiaorong Zhu Yuanyuan Li Fuzhen Yuan Honghua Sun Luaichang Gao Dianshuai |
| Institution: | Zhu Xiaorong1,2,Zhu Yuanyuan2,Li Fuzheng2,Yuan Honghua2,Sun Luaichang1,Gao Dianshuai2(1.Department of Preventive Veterinary,Veterinary Medicine College of Yangzhou University,Yangzhou 225009,2.Department of Neurobiology,Xuzhou Medical College,Xuzhou 221002,China) |
| Abstract: | Objective:To explore expression of calbindin-D28k(CaBP)In Vitro and the protective effect to the neurons by anti-apoptosis.Methods:The cDNA of CaBP was amplified from mice brain by RT-PCR.The pTH-CB(pcDNA3-TH-CB)plasmid was reconstructed and transfected in to MN-9D cells(Dopaminergic neurons hybridoma cells).The expression of the CaBP cDNA was detected by RT-PCR and immunoprecipitation,while the anti-apoptotic effect elicited by CaBP over expression was determined by Hoechst staining,caspase-3 activity dete... |
| Keywords: | calbindin D28k gene expression apoptosis mouse |
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