于志坚，艾启利，吕凯，张迪，朱亮

橙皮素对神经病理性疼痛大鼠坐骨神经的氧化应激以及炎性反应的影响

于志坚，艾启利，吕凯，张迪，朱亮

目的：探索橙皮素（HES）对坐骨神经压迫性损伤（PSNL）模型大鼠的镇痛作用及相关分子机制。方法制作大鼠PSNL模型。75只大鼠随机分为5组，15只/组，分别为假手术（sham）组、模型（model）组、HES低浓度（HES 10）组、HES中浓度（HES 20）组和HES高浓度（HES 40）组，分别给与HES 10 mg/kg、20 mg/kg、40 mg/kg灌胃给药，1次/日，连续21 d，sham组和model组给与等量生理盐水灌胃。分别于术前1 d和术后1、7、14、21 d测定大鼠后足热缩足反射潜伏期（TWL）及机械缩足反射阈值（MWT）；第21天取各组大鼠右侧坐骨神经，Elisa法检测氧化应激因子一氧化氮（NO）、炎症因子肿瘤坏死因子（TNF）-α、白介素（IL）-1β、IL-6和单核细胞趋化因子（MCP）-1含量和内皮型一氧化氮合酶（eNOS）活性；Western blot法检测eNOS蛋白表达水平，免疫组化法检测TNF-α、IL-1β的表达。结果造模后TWL和MWT显著下降（P＜0.05），HES治疗7、14、21 d后TWL及MWT明显升高，高于model组（P＜0.05）。Elisa结果显示，造模后，NO、TNF-α、IL-1β、IL-6、MCP-1含量及eNOS活性显著提高（P＜0.05）；HES治疗21 d后，NO、TNF-α、IL-1β、IL-6、MCP-1含量和eNOS活性明显降低，低于model组（P＜0.05）。Western结果显示，造模后，eNOS蛋白表达显著提高，HES可显著抑制eNOS蛋白表达，低于model组（P＜0.05）。免疫组化检测结果显示，HES治疗21 d后，TNF-α和IL-1β均显著低于model组（P＜0.05）。结论HES能缓解坐骨神经结扎引起的神经病理性疼痛，其分子机制可能与抑制坐骨神经氧化应激反应和炎症反应有关。

橙皮素；神经病理性疼痛；氧化应激；炎性反应；大鼠

AbstractObjective:To investigate the effects Hesperetin(HES)on relieving pain and the expression of oxidative stress factors and inflammatory factors in partial sciatic nerve ligation(PSNL)model rats.Methods:Seventy-five SD rats were randomly divided into five groups(n=15 respectively):sham group,model group,HES 10 group(10 mg/kg),HES 20 group(20 mg/kg)and HES 40 group(40 mg/kg).Thermal withdrawal latency(TWL)and mechanical withdrawal threshold(MWT)were tested 1 day before and 1,7,14,21 days after surgery.Twenty-one days after surgery,the rats were harvested.The content of nitric oxide(NO),tumor necrosis factor(TNF)-α,interleukin(IL)-1β,IL-6,monocyte chemotactic factor(MCP)-1 and the activity of endothelial nitric oxide synthase(eNOS)were detected by Elisa.The protein expression of eNOS was valued by western blot method and the protein expression of TNF-α and IL-1β were observed via immunohistochemical staining.Results:Compared with those in the sham group,TWL and MWT in model group were significantly down-regulated(P＜0.05).While 7,14,21 days after HES treatment,TWL and MWT in HES group were markedly up-regulated compared with those in the model group(P＜0.05).Data of Elisa showed that the content of NO,TNF-α,IL-1β,IL-6,MCP-1 and the activity of eNOS increased in model group(P＜0.05),which were significantly decreased after 21-day treatment of HES(P＜0.05).Data of western blot showed that the HES treatment significantly suppressed the protein expression of eNOS(P＜0.05).Data of immunohistochemical staining showed that HES treatment significantly suppressed the protein expression of TNF-α and IL-1β (P＜0.05).Conclusion:HES can relieve the neuropathic pain induced by sciatic nerve ligation,and the molecular mechanism may be related to the anti-oxidative stress and anti-inflammation.

神经病理性疼痛是指中枢和周围神经系统受到损伤引发的疼痛综合征[1]。在坐骨神经压迫性损伤（partial sciatic nerve ligation，PSNL）模型的病理生理过程中，氧化应激反应和炎症反应协同作用，使神经元兴奋性增加，引发神经病理性疼痛[2]。……