基于ERK/P38MAPK信号通路研究苗药金乌健骨方防治奥沙利铂诱导的神经病理性疼痛的作用机制

Mechanism of Miao Medicine Jinwu Jiangu Recipe(金乌健骨方)Preventing and Treating Oxaliplatin-Induced Neuropathic Pain Based on ERK/P38MAPK Signaling Pathway

目的基于细胞外调节蛋白激酶/丝裂原活化蛋白激酶p38(extracellular regulated kinase/p38 mitogen-activated kinaseERK/p38MAPK)信号通路探究苗药金乌健骨方防治奥沙利铂(oxaliplatin,OXA)诱导的神经病理性疼痛(chemotherapy-induced neuropathic pain,CINP)的作用机制。方法SPF级雌性SD大鼠40只,随机分为对照组、模型组、西药组、金乌健骨方组,每组10只。除对照组外,其余大鼠腹腔注射OXA 2 mg·kg^(-1)·d^(-1),连续给药5 d,建模慢性CINP模型。建模的同时,金乌健骨方组灌胃给药金乌健骨方24 g·kg^(-1)·d^(-1),西药组灌胃给药度洛西汀6.45 mg·kg^(-1)·d^(-1),对照组与模型组给予等量的生理盐水,连续给药15 d。分别于给药0、3、6、9、12、15 d检测大鼠机械痛阈值、热痛阈值,治疗结束后检测血清肿瘤坏死因子α(tumor necrosis factor-α,TNF-α),白细胞介素^(-1)β(interleukin^(-1)β,IL^(-1)β),白细胞介素-6(interleukin-6,IL-6)水平,并对脊髓L4-L5膨大神经组织进行病理检测,比较大鼠脊髓L4-L5背根神经节组织p-ERK1/2、p38MAPK、c-Jun氨基末端激酶(c-Jun N-terminal ki-nase,JNK)、细胞癌基因fos(Cellular oncogene fos,c-Fos)、环磷酸腺苷反应原件结合蛋白(cAMP response element binding protein,CREB)、核蛋白因子-κB(nucleoprotein factor-κB,Nf-κB)蛋白表达情况。结果对照组0~15 d机械痛阈值变化差异无统计学意义(P>0.05),第3天后,模型组械痛阈值开始下降,第9天后,模型组与金乌健骨方组、西药组机械痛阈值差异有统计学意义,金乌健骨方组、西药组机械痛阈值高于模型组,西药组高于金乌健骨方组(P<0.05);对照组0~15 d热痛阈值变化差异无统计学意义(P>0.05),模型组热痛阈值下降,第0天后,模型组与金乌健骨方组、西药组热痛阈值差异有统计学意义,金乌健骨方组、西药组热痛阈值高于模型组,西药组高于金乌健骨方组(P<0.05)。与对照组比较,模型组血清IL^(-1)β,IL-6及TNF-α水平升高,p-ERK1/2、p38MAPK、JNK、c-Fos、CREB、Nf-κB蛋白表达上调(P<0.05),与模型组比较,金乌健骨方组、西药组血清IL^(-1)β,IL-6及TNF-α水平降低,p-ERK1/2、p38MAPK、JNK、c-Fos、CREB、Nf-κB蛋白表达下调(P<0.05),与金乌健骨方组比较,西药组血清IL^(-1)β,IL-6及TNF-α水平降低,p-ERK1/2、p38MAPK、JNK、c-Fos、CREB、Nf-κB蛋白表达下调(P<0.05)。结论苗药金乌健骨方能够改善OXA诱导的神经病理性疼痛,其机制可能与调节ERK/p38MAPK通路有关。

Objective Based on the extracellular regulated protein kinase/mitogen-activated protein kinase p38(ERK/p38MAPK)signaling pathway,to explore the mechanism of Miao medicine Jinwu Jiangu Recipe(金乌健骨方)in the prevention and treatment of oxaliplatin(OXA)-induced neuropathic pain(CINP).Method Forty female SD rats of SPF grade were random⁃ly divided into control group,model group,western medicine group and Jinwu Jiangu Recipe group,with 10 rats in each group.Except for the control group,the other rats were intraperitoneally injected with OXA(2 mg·kg-1·d-1)for 5 days to establish a chronic CINP model.At the same time of modeling,Jinwu Jiangu Recipe group was intragastrically administered with Jinwu Jian⁃gu Recipe(24 g·kg-1·d-1),and the western medicine group was intragastrically administered with duloxetine 6.45(mg·kg-1·d-1).The control group and the model group were given the same amount of normal saline for 15 days.The mechanical pain threshold and thermal pain threshold of rats were detected on 0,3,6,9,12 and 15 d of administration,respectively.After treatment,serum levels of tumor necrosis factorα(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6)were measured.The enlarged nerve tissue was examined by pathology,and the protein expressions of p-ERK1/2,p38MAPK,c-Jun amino-ter⁃minal kinase(JNK),cellular oncogene fos(c-Fos),cyclic adenosine monophosphate response element binding protein(CREB)and nuclear protein factor-κB(Nf-κB)in the L4-L5 dorsal root ganglion tissue of the rat spinal cord were compared.Results There was no significant difference in the mechanical pain threshold changes in the control group from 0 to 15 d(P>0.05).Af⁃ter the third day,the mechanical pain threshold in the model group began to decrease.After the ninth day,there were statistically significant differences in the mechanical pain threshold among the model group,the Jinwu Jiangu Recipe group and the western medicine group(P<0.05).There was no significant difference in the thermal pain threshold changes in the control group from 0 to 15 d(P>0.05),while the thermal pain threshold in the model group decreased.The thermal pain thresholds in the Jinwu Jiangu Recipe group and the western medicine group were higher than those in the model group,and that in the western medicine group was higher than that in the Jinwu Jiangu Recipe group(P<0.05).Compared with those of the control group,the serum levels of IL-1β,IL-6 and TNF-αin the model group were increased,and the protein expressions of p-ERK1/2,p38MAPK,JNK,c-Fos,CREB and Nf-κB were up-regulated(P<0.05).Compared with those of the model group,the levels of serum IL-1β,IL-6 and TNF-αin the Jinwu Jiangu Recipe group and the western medicine group were decreased,and the protein expressions of p-ERK1/2,p38MAPK,JNK,c-Fos,CREB and Nf-κB were down-regulated(P<0.05).Compared with the Jinwu Jiangu Recipe group,the western medicine group decreased the serum levels of IL-1β,IL-6 and TNF-α,and the pro⁃tein expressions of p-ERK1/2,p38MAPK,JNK,c-Fos,CREB and Nf-κB decreased(P<0.05).Conclusion Miao medicine Jinwu Jiangu Recipe can improve OXA-induced neuropathic pain and its mechanism may be related to ERK/p38MAPK path⁃way.