摘要
目的研究TLR4/NF-κB信号通路在深低温停循环(DHCA)脑损伤发病机制中的作用。方法建立DHCA体外BV2细胞氧糖剥夺/复氧模型(OGD/R),随机分为正常组(C组)及OGD/R组(O组)。分别使用CCK-8和Western blot检测细胞活性以及TLR4、MyD88、NF-κB的蛋白水平,RT-PCR检测TLR4 mRNA表达水平,ELISA检测细胞培养上清TNF-α、IL-6表达量。结果 O组细胞活性比C组明显降低(P<0.05),O组TLR4、My D88及p-p65蛋白表达水平均较C组明显升高(P<0.05),O组TLR4 mRNA表达水平比C组显著增高(P<0.01),O组TNF-α及IL-6的表达量比C组增加,差异有统计学意义(P<0.01)。结论TLR4/NF-κB信号通路参与了OGD/R处理后BV2细胞的激活,并可能通过此途径参与DHCA脑损伤。
Objective To investigate the role of TLR4/NF-κB signal pathway in pathogenesis of brain inju-ry during deep hypothermia circulatory arrest（DHCA）. Methods BV2 microglia cells were subjected to oxygen-glucose deprivation/reoxygenation（OGD/R）,in vitro model for DHCA. The BV2 were randomly divided into thecontrol group（C group）and the experimental group（O group）. BV2 viability was determined by CCK-8 assay.TLR4 and its downstream signaling molecules,MyD88 and phosphorylated NF-κB（p-p65）expressions weredetected by Western blotting. TLR4 mRNA expression in BV2 microglial cells were determined by RT-PCR. Levelof interleukin-6（IL-6）and tumor necrosis factor alpha（TNF-α）in culture medium was detected by ELASA.Results Compared with the group C,BV2 microglia cell viability in experiment group was obviously weaker（P 0.05）. Expressions of TLR4,MyD88 and phosphorylated NF-κB（p-p65）from the experiment group increasedremarkedly than those from the group C（P 0.05）. TLR4 mRNA level was higher significantly in the group Othan in the group C（P 0.01）. Production of IL-6 and TNF-α in the group O were up-regulated apparentlycompared to the group C（P 0.01）. Conclusion TLR4/NF-κB signaling pathway contributed to activation of BV2 microglia cells treated by OGD/Reoxygenation,which was probably the exactly way that involved in pathogenesis ofbrain injury during deep hypothermia circulatory arrest.
出处
《实用医学杂志》
CAS
北大核心
2017年第20期3344-3347,共4页
The Journal of Practical Medicine
基金
国家自然科学基金资助项目(编号:81460290
81360287)
江西省自然科学基金资助项目(编号:20142BAB205041)
江西省教育厅科技资助项目(编号:GJJ12547
GJJ14695)
江西省卫生计生委科技计划资助项目(编号:20155427)
