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目的探讨亚低温对脂多糖(LPS)诱导的BV-2小胶质细胞Toll样受体4(TLR4)、核因子κB(NF-κB)及下游炎症因子表达的影响。方法体外培养的BV-2细胞,分为33℃和37℃条件下的生理盐水组和LPS组。利用实时定量PCR检测BV-2细胞中TLR4、NF-κB mRNA表达,用Western blot法检测BV-2细胞中TLR4、NF-κB蛋白的表达,利用ELISA检测培养液中肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)的水平。结果 LPS刺激BV-2细胞可导致炎性信号通路中的TLR4通路蛋白呈先升高后降低趋势,而通路下游的NF-κB蛋白表达呈持续上升趋势,炎性因子TNF-α、IL-1β的释放明显增加,亚低温明显抑制TLR4、NF-κB的mRNA和蛋白表达及炎症因子的释放。结论亚低温条件能够抑制BV-2细胞中LPS/TLR4通路,并抑制细胞因子TNF-α、IL-1β的表达。
Objective To investigate the effects of mild hypothermia on the expression of TLR4, NF-κB and downstream inflammatory cytokines in BV-2 microglia induced by lipopolysaccharide (LPS). Methods BV-2 cells cultured in vitro were divided into normal saline group and LPS group at 33 ℃ and 37 ℃. The expression of TLR4 and NF-κB mRNA in BV-2 cells was detected by real-time quantitative PCR. The expression of TLR4 and NF-κB in BV-2 cells was detected by Western blot. The levels of tumor necrosis factor- α), interleukin 1β (IL-1β). Results LPS stimulation of BV-2 cells resulted in the first increase and then decrease of TLR4 signaling pathway, while the downstream of NF-κB protein expression in the pathway continued to increase. The expressions of inflammatory cytokines TNF-α and IL-1β The release of TLR4 and NF-κB mRNA and protein and the release of inflammatory cytokines were significantly decreased after hypothermia. Conclusion Mild hypothermia can inhibit the LPS / TLR4 pathway in BV-2 cells and inhibit the expression of cytokines TNF-α and IL-1β.