基于c-Jun氨基末端激酶/p38丝裂原活化蛋白激酶信号通路探讨阿魏酸钠对偏头痛大鼠炎症反应的抑制作用

Inhibitory effect of sodium ferulate on inflammatory response in migraine rats based on c-Jun N-terminal kinase/p38 mitogen- activated protein kinase signaling pathway

目的 探讨阿魏酸钠(SF)通过调控JNK/p38 MAPK信号通路对偏头痛大鼠炎症反应的抑制作用。方法 腹腔注射硝酸甘油制备偏头痛大鼠模型,模型制作成功后随机分为模型组、SF低剂量(SF-L)组(50 mg/kg)、 SF高剂量(SF-H)组(100 mg/kg)、SF+JNK抑制剂(SF+SP600125)组(SF 100 mg/kg+SP600125 10 mg/kg)、 SF+JNK激活剂[SF+茴香霉素(AN)]组(SF 100 mg/kg+AN 5 mg/kg),每组12只,另取12只SD大鼠不做处理作为空白组。给药结束24 h后观察各组大鼠行为学变化,ELISA法检测血清中5-羟色胺(5-HT)、一氧化氮(NO)、肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)水平,TUNEL染色观察脑组织神经元凋亡情况,免疫组织化学法检测脑组织中TNF-α、 IL-6、降钙素基因相关肽(CGRP)表达,Western blotting法检测脑组织中JNK/p38 MAPK通路相关蛋白的表达。结果 与空白组比较,模型组大鼠挠头次数以及爬笼次数明显增加,神经元凋亡率显著升高;血清5-HT含量显著降低,NO、TNF-α和IL-6水平显著升高;脑组织中TNF-α、IL-6和CGRP表达以及磷酸化JNK(p-JNK)/JNK、磷酸化p38 MAPK(p-p38 MAPK)/p38 MAPK比值均显著升高(均P<0.05)。与模型组比较,SF-L组、SF-H组大鼠挠头次数及爬笼次数显著减少,神经元凋亡率显著降低;血清中5-HT含量显著升高,NO、TNF-α和IL-6水平显著降低;脑组织中TNF-α、IL-6和CGRP表达以及p-JNK/JNK、p-p38 MAPK/p38 MAPK比值均显著降低(均P<0.05)。与SF-H组比较,SF+SP600125组能够显著增强SF对偏头痛大鼠的保护作用;SF+AN组能够显著逆转SF对偏头痛大鼠的保护作用。结论 SF可能通过抑制JNK/p38 MAPK信号通路的表达,有效抑制偏头痛大鼠神经源性炎症反应,减少神经元凋亡,实现对偏头痛大鼠的保护作用。

Objective To explore the inhibitory effect of sodium ferulate(SF) on the inflammatory response in migraine rats by regulating the c-Jun N-terminal kinase(JNK)/p38 mitogen-activated protein kinase(p38 MAPK) signaling pathway. Methods The migraine rat model was prepared by intraperitoneal injection of nitroglycerin. After successful modeling, the rats were randomly grouped into model group, SF low dose(SF-L) group(50 mg/kg), SF high dose(SF-H) group(100 mg/kg), SF+JNK inhibitor(SF + SP600125) group(SF 100 mg/kg +SP600125 10 mg/kg), and SF+JNK activator [SF + anisomycin(AN)] group(SF 100 mg/kg +AN 5 mg/kg), 12 in each group, another 12 SD rats without treatment were taken as blank group. The behavioral changes of the rats in each group were observed 24 hours after the administration, the levels of 5-hydroxytryptamine(5-HT), nitric oxide(NO), tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) in serum were detected by ELISA, the neuronal apoptosis in brain tissue was observed by TUNEL staining, immunohistochemistry was used to evaluate the expressions of TNF-α, IL-6 and calcitonin gene-related peptide(CGRP) in brain tissue, Western blotting was used to detect the expressions of JNK/p38 MAPK pathway-related proteins in brain tissue. Results Compared with the blank group, the number of times of scratching the head and climbing the cage of the rats in the model group increased significantly, and the apoptosis rate of neurons increased significantly;the content of 5-HT in serum decreased significantly, and the levels of NO, TNF-α and IL-6 increased significantly;the expressions of TNF-α, IL-6 and CGRP, and the ratios of phosphorylated JNK(p-JNK)/JNK and phosphorylated p38 MAPK(p-p38 MAPK)/p38 MAPK in brain tissue obviously increased(all P<0.05). Compared with the model group, the number of times of scratching the head and the times of climbing the cage of the rats in the SF-L group and the SF-H group reduced significantly, and the neuron apoptosis rate reduced significantly;the content of 5-HT in serum increased significantly, and the levels of NO, TNF-α and IL-6 decreased significantly;the expressions of TNF-α, IL-6 and CGRP, and the ratios of p-JNK/JNK and p-p38 MAPK/p38 MAPK in brain tissue obviously decreased(all P<0.05). Compared with SF-H group, the protective effect of SF on migraine rats in SF+SP600125 group enhanced significantly;the protective effect of SF on migraine rats in the SF+AN group reversed significantly. Conclusion SF may inhibit the expression of JNK/p38 MAPK signaling pathway, effectively inhibit neurogenic inflammatory response in migraine rats, reduce neuronal apoptosis, and achieve a protective effect on migraine rats.