炎症因子对原代培养海马神经元CLC-3表达的影响

Effects of inflammatory cytokines on the change of CLC-3 expression in primary cultured hippocampal neurons

目的:研究不同浓度TNF-α、IL-1β、TGF-β刺激对原代培养海马神经元CLC-3(voltage-gated chloride channel 3)mRNA及蛋白质水平的影响。方法:取原代培养8 d的大鼠海马神经元,随机分为对照组、TNF-α组、IL-1β组和TGF-β组。TNF-α、IL-1和TGF-β组分别加入加入含有浓度为10 ng/ml的相应炎症因子培养液,每组分别在孵育6、12、24 h后分别收集细胞提取总RNA和蛋白采用实时定量PCR、Western Blot技术检测CLC-3 mRNA及蛋白水平的表达。结果:TNF-α、IL-1β处理12 h后培养海马神经元CLC-3在mRNA及蛋白水平开始上调,高峰持续从12 h至24 h(P<0.05)。TNF-α刺激作用强于IL-1β。TGF-β处理海马神经元CLC-3 mRNA在6 h后即开始升高(P<0.05),但在孵育6 h到24 h时下降至正常对平(P>0.05)。结论:TNF-α、IL-1β、TGF-β可能通过刺激海马神经元CLC-3表达增加增强神经元存活能力。

Objective： To explore the effects TNF-α,IL-1β,TGF-β on the expression of voltage-gated chloride channel3（ CLC-3） mRNA and protein in primary cultured rat hippocampal neurons. Methods： Hippocampal neurons were cultured for 8 days and then divided into control,TNF-α,IL-1β and TGF-β groups. Cells in TNF-α,IL-1β and TGF-βgroups were treated with 10 ng / ml inflammatory cytokines respectively. The expression of CLC-3 at mRNA and protein levels were measured at time points of 6,12 and 24 h after coculture by real time PCR and Western Blot. Results： CLC-3 mRNA and protein were significantly increased at 12 h and kept maximal level from 12 h to 24 h in TNF-α and IL-1βgroups（ P 〈 0. 05）. TNF-α has stronger stimulation effect than IL-1β. CLC-3 expression was strengthened at 6 h in TGF-β group（ P 〈 0. 05） but decreased to normal level at time points of 12 h and 24 h（ P 〉 0. 05）. Conclusion： TNF-α,IL-1β and TGF-β may enhance the viability of cells by raising the CLC-3 expression.