IL-1β促进Schwann细胞增殖及分泌NGF的研究  

Study on the IL-1β promote the proliferation of Schwann cells and secreting NGF

作　　者：于巧莲[1] 秦建强[1] 余磊[1] 邱小忠[1] 廖华[1] 王齐[1] 朴英杰[1] 

机构地区：[1]南方医科大学解剖学教研室广东省组织构建与检测重点实验室,广州510515

出　　处：《神经解剖学杂志》2008年第3期319-324,共6页Chinese Journal of Neuroanatomy

基　　金：广东省科技计划专项(2003A3020101);国家自然科学基金(No39970833)资助项目

摘　　要：为观察白介素-1β(IL-1β)对Schwann细胞增殖及分泌神经生长因子(NGF)的影响,本研究取3d龄的大鼠坐骨神经纯化培养Schwann细胞,然后分4组进行处理。处理方法为:A组加入含10%胎牛血清的DMEM/F12培养基,B组仅在纯化后第1d向培养基中加入2.0ng/mlIL-1β,C组每天向培养基中加入2.0ng/mlIL-1β,D组隔天向培养基中加入2.0ng/mlIL-1β,分别培养5d。倒置显微镜下观察Schwann细胞的形态,用抗S-100蛋白免疫组化鉴定Schwann细胞,经流式细胞仪(FCM)检测细胞的分裂增殖情况,用反转录-聚合酶链式反应(RT-PCR)方法检测NGFmRNA表达的变化,酶联免疫吸附分析法(ELISA)检测细胞培养基中NGF蛋白的表达量。结果显示:D组Schwann细胞增殖率、NGFmRNA合成量和NGF蛋白表达量均显著高于其他三组(P<0.05)。此结果说明IL-1β能够促进Schwann细胞的分裂增殖,并且促进胞内NGFmRNA的合成及胞外NGF的分泌,其作用持续时间大约为2d。To observe the effect of Schwann cells proliferation and secreting nerve growth factor （NGF） by interleukin-1β （ IL-1β）, we purified and cultured Schwann cells of sciatic nerves from 3 days old rats. The cells were classified four groups to be treated. Group A ： DMEM/F12 contained 10% calf blood serum was added into the culture medium; group B： IL-1β （2.0 ng/ml） was added only in the first day after the purification; group C： IL-1β （2.0 ng/ml） added every day; group D： IL-1β （2.0 ng/ml） was added every other day. Four groups were cultured for 5 days, then the morphology of Schwann cells was observed with inverted microscope. We evaluated Schwann cells by immunecytechemically staining with antibody against S-100 protein. The proliferation of Schwann cells was detected by flow cytometry （FCM）. The expression of NGF mRNA and its protein were analyzed by reverse transcription-polymerase chain reaction （RT-PCR） and enzyme-linked immunoadsorbent assay （ ELISA ）, respectively. The results showed that there were significant differences between group D and other three groups with respect to the level of Schwann cells proliferation, the synthesis of NGF mRNA and expression of its protein （ P 〈 0.05 ）. The level of these parameters of group D were obviously higher than that of groups A, B and C. The results suggest that IL-1β can promote the proliferation of Schwann cells, and the synthesis of NGF mRNA in the cell and the extraeellular secretion of NGF and these effects last about two days.

关 键 词：白介素-1Β SCHWANN细胞 神经生长因子 周围神经 大鼠 

分 类 号：R392[医药卫生—免疫学]