Sev对低氧缺糖诱导的脑神经元细胞损伤的影响

Sevoflurane regulates TLR4/NF-κB signaling pathway and inhibits the protective effect of hypoxia and hypoglycemia on neuronal damage

目的 研究七氟醚(Sev)调控Toll样受体4 (TLR4)/核转录因子κB (NF-κB)信号通路抑制低氧缺糖对脑神经元细胞损伤的保护作用。方法 将大鼠皮质神经元细胞分为对照组、模型组、1.0MAC和2.0 MAC七氟醚组;细胞计数试剂盒(CCK-8)法、流式细胞术检测细胞增殖和凋亡。试剂盒检测丙二醛(MDA)含量以及超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性。酶联免疫吸附测定法(ELISA)检测肿瘤坏死因子(TNF-α)、白细胞介素-6 (IL-6)、白细胞介素-8 (IL-8)水平;蛋白印迹法(Western blot)检测Bcl-2相关X蛋白(Bax)、B细胞淋巴瘤/白血病-2 (Bcl-2)、TLR4、磷酸化核因子κB抑制蛋白α (p-IκBα)和核因子κB抑制蛋白α (IκBα)蛋白表达。结果 与对照组比较,模型组细胞存活率、SOD、CAT活性降低(均P<0.05),凋亡率、MDA含量、TNF-α、IL-6、IL-8水平升高(P<0.05),TLR4、p-IκBα蛋白表达升高;与模型组比较,Sev显著升高细胞存活率、SOD、CAT活性(P<0.05),显著降低细胞凋亡率、MDA含量、TNF-α、IL-6、IL-8水平(P<0.05),TLR4、p-IκBα蛋白表达降低(P<0.05)。结论Sev可减轻低氧缺糖诱导的脑神经元细胞损伤,其机制可能与抑制TLR4/NF-κB信号通路有关。

Objective To study the protective effect of sevoflurane(Sev)in regulating the Toll-like receptor4(TLR4)/nuclear transcription factorκB(NF-κB)signaling pathway and inhibiting hypoxia and hypoglycemia against neuronal injury.Methods The cortical neurons of rats were divided into control group,model group,1.0 MAC and 2.0MAC sevoflurane groups.Cell counting kit(CCK-8)method,flow cytometry to detect cell proliferation and apoptosis.The content of malondialdehyde(MDA)and the activity of superoxide dismutase(SOD)and catalase(CAT)were detected by kit.The levels of tumor necrosis factor(TNF-α),interleukin-6(IL-6)and interleukin-8(IL-8)were detected by enzyme-linked immunosorbent assay(ELISA).Western blot was used to detect Bcl-2-related X protein(Bax),B-cell lymphoma/leukemia-2(Bcl-2),TLR4,phosphorylated nuclear factorκB inhibitory proteinα(p-ⅠκBα)and nuclear factorκB inhibitory protein alpha(ⅠκBα)protein expression.Results Compared with the control group,the cell survival rate,SOD and CAT activities in the model group decreased(~(all)P<0.05),while the apoptosis rate,MDA content,TNF-α,IL-6 and IL-8 levels increased(P<0.05),the expression of TLR4 and p-IκB a protein increased;Compared with the model group,sevoflurane significantly increased cell survival rate,SOD,and CAT activity(P<0.05),and significantly decreased cell apoptosis rate,MDA content,TNF-α,IL-6,and IL-8 levels(P<0.05),the expression of TLR4 and p-ⅠκBαprotein decreased(P<0.05).Conclusion Sev can alleviate the damage of brain nerve cells induced by hypoxia and glucose deprivation,and its mechanism may be related to the inhibition of TLR4/NF-κB signaling pathway.