高压氧治疗对癫痫大鼠血清白细胞介素1β、白细胞介素2、白细胞介素8、肿瘤坏死因子α水平及海马神经元Bax及Bcl-2表达的影响

Effects of hyperbaric oxygen therapy on serum IL-1β,IL-2,IL-8,TNF-αlevels and expression of Bax and Bcl-2 in hippocampal neurons in epileptic rats

目的:探究高压氧治疗对癫痫大鼠血清白细胞介素(IL)1β、IL-2、IL-8、肿瘤坏死因子α(TNF-α)的表达及海马神经元细胞凋亡基因表达的影响。方法:采用随机数字法将36只成年健康SD大鼠进行编号,1~12号实验动物为空白对照组,剩余动物接受匹罗卡品建立癫痫模型,根据Racine评分作为完成标准,随机选取12只接受高压氧治疗,利用ELISA法检测实验动物血清IL-1β、IL-2、IL-8、TNF-α表达水平,利用免疫印迹和免疫荧光染色检测实验动物海马神经元细胞凋亡蛋白Bax和Bcl-2表达。结果:与空白对照组相比,癫痫模型组大鼠血清IL-1β、IL-8、TNF-α表达水平均显著升高,IL-2表达水平明显降低;经过高压氧治疗后,与癫痫模型组相比,实验大鼠血清IL-1β、IL-8、TNF-α表达水平显著降低,IL-2表达水平明显升高。免疫印迹和免疫荧光染色结果显示,与空白对照组相比,癫痫模型组促凋亡蛋白Bax表达显著升高,而抗凋亡蛋白Bcl-2表达显著下降,经过高压氧治疗后,Bax蛋白水平显著下调,而Bcl-2蛋白水平显著上调。结论:高压氧治疗可减少癫痫大鼠血清IL-1β、IL-8、TNF-α的表达,增加IL-2的表达,Bax蛋白水平显著下调而Bcl-2蛋白水平显著上调,因此本研究可作为高压氧治疗癫痫病作用机制之一。

Objective:To explore the effect of hyperbaric oxygen therapy on the levels of serum interleukin(IL)-1β,IL-2,IL-8,tumor necrosis factor-α(TNF-α)and apoptosis gene of hippocampal neurons in epileptic rats.Methods:Thirty-six healthy adult SD rats were numbered by using random numbering.Rats numbered from 1 to 12 were classified into the blank control group.The pilocarpine method was used among the rest of the experimental animals to establish the epilepsy model.According to the Racine score as the completion standard,12 rats were randomly selected to receive hyperbaric oxygen treatment.The expression levels of IL-1β,IL-2,IL-8 and TNF-αin serum were detected by ELISA.The expression levels of Bax and Bcl-2 in hippocampal neurons were detected by Western blotting and immunofluorescence staining.Results:Compared with the blank control group,the serum IL-1β,IL-8,TNF-αexpression levels of rats in the model group were significantly elevated,and IL-2 expression decreased obviously.After hyperbaric oxygen therapy intervention,compared with epilepsy model group,expression of the serum IL-1β,IL-8,TNF-αin experimental rats decreased significantly,the IL-2 expression increased significantly.Western blotting and immunofluorescence staining of hippocampal neuron for detecting apoptosis factor Bax and Bcl-2 found that the expression of apoptosis promoting protein Bax increased significantly compared with the blank control group,but the expression of antiapoptotic protein(Bcl-2)decreased significantly in the epilepsy model group.After hyperbaric oxygen therapy intervention,Bax protein levels were significantly lower and the Bcl-2 protein levels rose significantly.Conclusion:Hyperbaric oxygen therapy can significantly reduce the expressi on of IL-1β,IL-8 and TNF-αand increase the expression of IL-2 in epileptic rats.Meanwhile,after hyperbaric oxygen therapy intervention,Bax protein level is significantly down-regulated while Bcl-2 protein level is significantly up-regulated.Therefore,this study can be used as one of the mechanisms of hyperbaric oxygen therapy for epilepsy.