白介素-1β对原代培养海马神经元毒性的研究

目的 探讨IL-β对体外培养的海马神经元活性及合成和分泌脑源性神经营养因子(BDNF)的影响.方法 采用SD新生鼠进行海马神经元原代培养.培养7d的细胞以细胞免疫化学法枪测神经元特异性烯醇化酶(NSE)的表达,计算阳性细胞率.实验分IL-β为0ng/ml的正常组和IL-β为5 ng/ml、10ng/ml、20ng/ml 3个实验组,进行海马神经元无血清培养48 h.采用甲基噻唑基四唑(MTT)试验检测细胞活性,ELISA法检测培养上清BDNF的含量.结果 NSE细胞免疫化学鉴定显示阳性细胞达到90%以上;IL-1β降低体外培养的海马神经元活性,各剂量组按浓度在590nm波长处检测OD值为(0.5585±0.2407; 0.4295±0.1401; 0.4191±0.1050),与正常组(0.9462±0.2972)比较,差异具有显著性(分别为P<0.05,P<0.01,P<0.01 ANOVA);IL-1β降低体外培养的海马神经元合成和分泌BDNF水平,各剂量组显著BDNF含量分别是(8.65±0.71)pg/ml,(8.90±0.35)pg/ml,(7.90±0.35)pg/ml],与正常组(12.40±1.77)pg/ml比较差异具有显著性(均P<0.05 ANOVA).结论 IL-1β可导致体外培养海马原代神经元的损伤,其机制可能与海马神经元合成和分泌BDNF的减少有关.

Neurotoxicity of interleukine-1 beta on hippocampus neuron in vitro

Objective To examine cell viability and brain-derived neurotrophic factor( BDNF) concentration in the effect of interleukine-1 β(3 (IL-1β) on hippocampus neuron primary cell culture. Methods Hippocampus were dissected and cultured from new-born SD rats,the cultured cells were fixed on 7 days and immunocyto-chemically stained with neuronal specific enolase (NSE) antibody. Interleukine-1 beta IL-1β 0ng,5ng/ml, 10ng/ ml,20ng /ml was applied to no serum cultured neurons for 48 h. The cell proliferation and viability was determined by methyl thiagolyl tetragolium( MTT) assay, BDNF concentrations in the supernatant was determined by ELISA kits. Results Immunocytochemically stained with NSE antibody results indicated that above 90% of cell isolated from hippocampus of 7 d rats greed well and expressed NSE immunoreativity. The cell viability decreased follow with the dose of IL-1β, the optical density of each dose detected at 590 nm wave were(0.9462 ± 0.2972;0.5585 ± 0.2407; 0.4295 ±0.1401;0.4191 ±0.1050), compared with normal group ( P<0.05, P<0.01 , P<0.01). BDNF concentrations in the supernatant was decreased compared to control group,and the contents of BDNF in each group were(12.4±1.77)pg/ml; (8.65±0.71)pg/ml;(8.9±0.35)pg/ml;(7.9±0.35)pg/ml], compared with normal group ( P<0.05, P<0.05, P<0.05). Conclusion Neurotoxicity of IL-1β on hippocampus neuron in vitro might via decreased synthesis and secretion of BDNF.