基于EphrinBs/EphBs通路探讨桃红四物汤治疗带状疱疹后遗神经痛大鼠模型的作用机制  

Study on the mechanism of Taohong Siwu Decoction in the treatment of postherpetic neuralgia in rats based on EphrinBs/EphBs pathway

作　　者：徐俊涛[1] 王丽[1] 王莹[1] 马飞[2] 方玉甫[1] XU Jun-tao;WANG Li;WANG Ying;MA Fei;FANG Yu-fu(Department of Dermatology,Henan Traditional Chinese Medicine Hospital,Zhengzhou 450002,China;Department of Paediatrics,the Sixth Affiliated Hospital of Sun Yat-sen University)

机构地区：[1]河南省中医院皮肤科,450002 [2]中山大学附属第六医院儿科

出　　处：《天津医药》2021年第2期147-152,共6页Tianjin Medical Journal

基　　金：河南省中医药科学研究专项课题(2019JDZX052)。

摘　　要：目的观察桃红四物汤对带状疱疹后遗神经痛(PHN)大鼠模型的影响,探讨EphrinBs/EphBs通路在其中的作用机制。方法60只SD大鼠采用随机数字表法分为空白组、模型组(采用水痘-带状疱疹病毒慢性感染法构建PHN模型)、阳性组[建模后以普瑞巴林27 mg/(kg·d)灌胃]、治疗组[建模后以桃红四物汤5 g/(kg·d)灌胃],治疗+EphB2-Fc组[建模第3天鞘内注射EphB2-Fc 5μg后以桃红四物汤5 g/(kg·d)灌胃]和治疗+EphrinB2-Fc组[建模第3天鞘内注射EphrinB2-Fc 5μg后以桃红四物汤5 g/(kg·d)灌胃]。采用行为学法分别在给药后1 d、7 d和14 d时检测大鼠的机械痛阈值,酶联免疫吸附测定(ELISA)法检测大鼠脊髓组织中白细胞介素(IL)-1β和肿瘤坏死因子(TNF)-α水平,原位末端转移酶标记技术(TUNEL)法检测脊髓神经细胞凋亡,荧光法检测脊髓组织的半胱氨酸天冬氨酸蛋白酶3(Caspase-3)活性,蛋白免疫印迹法(Western blot)检测EphrinBs/EphBs通路相关蛋白EphrinB2和EphB2的表达。结果与空白组比较,建模后大鼠机械痛阈值明显降低,而大鼠脊髓中IL-1β、TNF-α水平、脊髓神经细胞凋亡指数、Caspase-3活性以及EphrinB2、EphB2蛋白表达水平均明显升高(P<0.05);与模型组比较,给予桃红四物汤治疗的大鼠在给药后7 d和14 d时机械痛阈值均明显升高(P<0.05),而大鼠脊髓中IL-1β、TNF-α水平、脊髓神经细胞凋亡指数、Caspase-3活性以及EphrinB2、EphB2蛋白表达水平均明显降低(P<0.05)。给予EphBs阻滞剂EphB2-Fc后大鼠脊髓组织中EphB2蛋白表达水平明显低于治疗组(P<0.05)。以EphBs激动剂EphrinB2-Fc作用后大鼠脊髓组织中EphrinB2蛋白表达水平明显低于治疗组,而EphB2蛋白表达水平明显高于治疗组(P<0.05)。结论桃红四物汤可能通过抑制EphrinBs/EphBs通路减少炎性因子的释放和脊髓神经细胞凋亡,对PHN大鼠发挥镇痛作用。Objective To observe the effect of Taohong Siwu Decoction on postherpetic neuralgia(PHN)rats and to explore the role of EphrinBs/EphBs pathway in it.Methods A total of 60 SD rats were divided into blank group,model group(PHN model was established by chronic infection of varicella-zoster virus),positive group(27 mg·kg-1·d-1 of pregabalin was given by intragastric administration after modeling),treatment group(5 g·kg-1·d-1 of Taohong Siwu Decoction was given by intragastric administration after modeling),treatment+EphB2-Fc group(after intrathecal injection of 5μg EphB2-Fc on the 3rd day of modeling,5 g·kg-1·d-1 of Taohong Siwu Decoction was given by intragastric administration)and treatment+EphrinB2-Fc group(after intrathecal injection of 5μg EphrinB2-Fc on the 3rd day of modeling,5 g·kg-1·d-1 of Taohong Siwu Decoction was given by intragastric administration)using random number table.The mechanical pain threshold of rats was measured by behavioral method at the 1st,7th and 14th days after administration.The levels of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)in the spinal cord of rats were measured by enzyme-linked immunosorbent assay(ELISA).Terminal dexynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)was used to detect apoptosis of spinal cord neurons.Fluorescence was used to detect the cysteinyl aspartate specific proteinase 3(Caspase-3)activity in spinal cord.Western blot assay was used to detect the expressions of EphrinB2 and EphB2 in EphrinBs/EphBs pathway.Results Compared with the blank group,the mechanical pain threshold was significantly lower in model group,while the levels of IL-1β,TNF-α,apoptosis index of spinal nerve cells,Caspase-3 activity and the expression levels of EphrinB2 and EphB2 proteins were significantly higher(P<0.05).Compared with the model group,the mechanical pain threshold of the rats treated with Taohong Siwu Decoction was significantly higher on the 7th day and 14th day after administration(P<0.05).However,the levels of IL-1β,TNF-αin the

关 键 词：神经痛 带状疱疹后 炎症 细胞凋亡 受体 Eph家族 膜附着型配体类 桃红四物汤 EphrinBs/EphBs通路 

分 类 号：R275[医药卫生—中医皮科]