摘要
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 impact of mild hypothermia on the lipopolysaccharide （LPS）-induced expressions of Toll-like receptor 4 （ TLR4 ）, nuclear factor κB （ NF-κB ） and downstream inflammatory cytokines in BV-2 microglias. Methods BV-2 cells cultured in vitro were divided into 33℃-PBS, 33℃-LPS, 37℃-PBS and 37℃-LPS groups. Real-time quantitative PCR was performed to detect the mRNA expressions of TLR4 and NF-κB in BV-2 cells, Western blotting was performed to detect the protein expressions of TLR4 and NF-κB in BV-2 cells, and ELISA to detect the levels of tumor necrosis factor α （TNF-α） and interleukin-1β （IL-1β） in the culture medium. Results After LPS stimulated BV-2 cells, TLR4 pathway protein exhibited the trend of increasing firstly and decreasing afterwards, while the expression of NF-κB protein in the pathway downstream continued to increase, and the release of inflammatory cytokines TNF-α and IL-1β were significantly promoted. Mild hypothermia could significantly inhibit the transcription and expressions of TLR4 and NF-κB as well as the release of inflammatory cytokines. Conclusion The mild hypothermia could inhibit the LPS/TLP4 pathway in BV-2 cells, and depress the expressions of TNF-α and IL-1β.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2015年第11期1479-1482,1487,共5页
Chinese Journal of Cellular and Molecular Immunology
基金
安徽省科技攻关项目(1301042213)
