LPS预激神经胶质细胞培养上清对神经元的影响  

Effects of neuroglia cultured supernatants with LPS-priming on neurons

作　　者：谢泽锋[1] 苏芸 蒋治武[2] 许燕璇[2] 辛岗[2] 李康生[2] 

机构地区：[1]汕头大学医学院第一附属医学院,广东汕头515041 [2]汕头大学医学院微生物学与免疫学教研室,广东汕头515041

出　　处：《中国热带医学》2012年第6期652-655,共4页China Tropical Medicine

基　　金：国家自然科学基金项目(No.81001340;30771988);广东省医学科研基金项目(No.B2010211);汕头市科技计划(No.2009-70)

摘　　要：目的探讨神经胶质细胞经过脂多糖(Lipopolyssacride,LPS)预激后,其培养上清对神经元功能的影响。方法体外培养神经元与神经胶质细胞,免疫荧光法进行纯度鉴定。神经胶质细胞分为4组:未刺激对照组、0.01μg/ml LPS单次刺激组、1μg/ml LPS单次刺激组、预刺激组(先采用0.01μg/ml LPS刺激18h后,再用1μg/mlLPS刺激24h)。经过相应处理后收获各组条件培养上清,分别加入到神经元中,与神经元共同孵育24h后收集细胞与培养上清。采用CCK-8试剂盒与ELISA法检测神经元的存活率和分泌TNF-α、IL-6水平。结果星形胶质细胞预刺激组条件培养上清作用于神经元后,细胞存活率下降,与0.01μg/ml、1μg/ml LPS单次刺激组比差异均具有统计学意义(P<0.05);小胶质细胞各组条件培养上清作用于神经元后,细胞存活率变化不明显(P均>0.05);在星形胶质细胞各刺激组上清作用下,神经元产生TNF-α的水平,各LPS处理组培养上清刺激下水平升高,其中1μg/ml LPS单次刺激组升高明显,与对照组和0.01μg/ml LPS单次刺激组比差异具有统计学意义(P均<0.05);产生IL-6的水平亦升高,与对照组比亦均具有统计学差异(P<0.05或P<0.01);并且,预刺激组较1μg/mlLPS单次刺激组上清作用后,产生IL-6水平较低(P<0.05)。各组小胶质细胞条件培养上清作用于神经元后,产生TNF-α的水平,预刺激组水平升高明显,与对照组比差异具有统计学意义(P<0.01);对于IL-6水平,变化趋势同TNF-α,但各组间差异无统计学意义(P>0.0.5)。结论 LPS预激参与重新调配了神经胶质细胞分泌活性介质的作用,培养上清中这些分子相互制约或是协同,对神经元产生有可能是保护或是损伤效应。Objective To study the effects of glia cultured supernatants with LPS-priming on neurons＇ viability and secretion activity.Methods Neurons and glia（mainly referring to astrocytes and microglia）were cultured in vitro and identified by immunofluorescence.Thereafter,these glial cells were divided into 4 groups：LPS-primed group,0.01μg/ml of LPS treated group,1μg/ml of LPS treated group and non-treated group.The LPS-primed group was preconditioned with 0.01μg/ml of LPS for 18 hours and re-retreated with a higher dose of LPS（1μg/ml LPS）for 24 hours.Then all cultured supernatants of 4 groups were collected and were used to treat the neurons for 24 hours in vitro.The cytotoxicity and secretion ability of proinflammatory cytokines were determined by CCK-8 kit and ELISA respectively.Results The cytotocixity of neurons was decreased by the treatment with the astrocyte supernatant of LPS-primed group,there was significant difference statistically if compared with 0.01μg/ml and 1ug/ml of LPS groups（P0.05;P0.05）.The cytotocixity showed no profound variation after the treatment with the microglia supernatants of 4 groups（P0.05）.As for TNF-α levels,it was increased by the treatment with the astrocyte supernatant of LPS-primed group,0.01μg/ml of LPS treated group,1μg/ml of LPS treated group,particularly in 1ug/ml of LPS treated group,statistical difference was found when compared with 0.01μg/ml of LPS treated group and non-treated group（P0.05;P0.05）;IL-6 levels were increased after the treatment with the astrocyte supernatants of LPS-primed group,0.01μg/ml of LPS treated group,1ug/ml of LPS treated group（P0.05）.Furthermore,after the treatment with the microglia supernatants of LPS-primed group,TNF-α level was increased profoundly when compared with non-treated group（P0.01）.Though IL-6 levels exhibited an increasing trend,no significant difference was found in statistically between groups（P0.05）.Conclusion There were diverse effects（ncluding protection or injury） of cultured super

关 键 词：神经元 脂多糖 神经胶质细胞 

分 类 号：R-33[医药卫生]