大鼠骨髓基质细胞分泌胶质细胞源性神经营养因子的研究  

Expression of mRNAs and protein for glial cell line-derived neurotrophic factors in bone marrow stromal cells of SD rat

作　　者：叶民[1] 陈生弟[1] 戚晨[1] 陆国强[1] 梁梁[1] 徐洁懿[1] 

机构地区：[1]上海第二医科大学附属瑞金医院神经科,帕金森病诊疗与研究中心上海200025

出　　处：《神经科学通报》2005年第1期23-27,共5页Neuroscience Bulletin

基　　金：国家重点基础研究规划"脑功能和脑重大疾病的基础研究"(G1999054008);国家自然科学基金(39970263;30171025);上海市卫生系统百名跨世纪优秀学科带头人培养计划(97BR001)

摘　　要：目的研究骨髓基质细胞(BMSCs)能否分泌胶质细胞源性神经营养因子(GDNF)及其分泌规律,为进一步探讨BMSCs治疗帕金森病(PD)大鼠模型的作用机制提供实验依据。方法取较高纯度的BMSCs进行培养,通过RT-PCR和ELISA法,分别从mRNA和细胞蛋白水平测定体外培养BMSCs的上清液和细胞蛋白中GDNF的含量。结果通过RTPCR法检测结果显示在mRNA水平,BMSCs中有GDNF的表达。在蛋白水平,采用ELISA法在培养第3天以后的培养液和细胞蛋白中均可检测到GDNF,并随着时间的延长,培养液和细胞蛋白中GDNF浓度明显升高。结论BMSCs具有分泌GDNF的能力,细胞分泌可能受周围环境和自身生长状况的影响,其分泌是非持续性的。GDNF浓度的升高不仅与BMSCs细胞数量增加有关,还与细胞本身分泌能力的增强有关。Objective To investigate the capability of bone marrow stromal cells (BMSCs) of SD rats in the secretion of glial cell line-derived neurotrophic factors(GDNF), and to provide an experimental basis for the mechanism of the treatment of BMSCs in Parkinson’s disease. Methods BMSCs were harvested from SD rats and cultured in DMEM supplemented with 20% fetal bovine serum. At passage 2 the total RNA was isolated using TriZol reactive. The expression of GDNF mRNA and GDNF protein were detected by the assays of RT-PCR and ELISA. Results The results showed that the mRNA and protein of GDNF were detected in culture supernatant and cultured cells after 3 d on of culture. GDNF in culture supernatant and cells increased gradually in 3, 7 and 10 d of culture. Conclusion Rat BMSCs have potential in the secretion of GDNF without continuousness, which is determined by the inner environment and self-growth status. GDNF elevation during cell culture was attributed to not only expansion of the number of cells, but also enhanced ability of BMSCs in secreting GDNF.

关 键 词：骨髓基质细胞 胶质细胞源性神经营养因子 逆转录酶-聚合酶链反应 酶联免疫吸附试验 治疗 帕金森病 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]