加巴喷丁有效缓解带状疱疹后神经痛大鼠痛敏并调控CACNA2D1的表达  

Gabapentin effectively alleviates pain sensitization and regulates CACNA2D1 expression in postherpetic neuralgia rats

作　　者：龙兴震 余胜丽[2] 马尊峰 霍文耀 孙蔺波 张洋 白雪[2] LONG Xing-zhen;YU Sheng-li;MA Zun-feng;HUO Wen-yao;SUN Lin-bo;ZHANG Yang;BAI Xue(The First Affiliated Hospital of Guizhou University of Traditional Chinese Medicine,Guiyang 550001,China;Guizhou Provincial People′s Hospital,Guiyang 550002,China)

机构地区：[1]贵州中医药大学第一附属医院,贵州贵阳550001 [2]贵州省人民医院,贵州贵阳550002

出　　处：《中国药理学通报》2023年第5期903-909,共7页Chinese Pharmacological Bulletin

基　　金：贵州省人民医院青年基金(No GZSYQN[2017]14);贵州省科技支撑项目(No QKHZC[2018]2801);国家自然科学基金资助项目(No 81960681)。

摘　　要：目的明确加巴喷丁治疗带状疱疹后神经痛(postherpetic neuralgia,PHN)的分子作用靶点。方法通过网络药理学和分子对接的方法分析加巴喷丁治疗PHN的分子靶点并通过共沉淀试验验证。制备PHN大鼠模型,随机数字表法分为正常对照组、疾病模型组、模型组+50 mg·kg^(-1)加巴喷丁组、模型组+100 mg·kg^(-1)加巴喷丁组、模型组+200 mg·kg^(-1)加巴喷丁组,每组9只,并在不同时间点测定大鼠痛觉行为学;免疫荧光、Western blot和qPCR测定大鼠脊髓组织CACNA2D1、Bax和Bcl-2 mRNA及其蛋白表达。结果通过网络药理学、分子对接和共沉淀试验证明,CACNA2D1是加巴喷丁的靶基因,二者能够稳定结合。各组大鼠在造模后机械痛阈和热疼痛阈明显降低,GABA细胞凋亡数量明显增加;分别腹腔注射50、100和200 mg·kg^(-1)加巴喷丁后,大鼠的机械痛阈和热疼痛阈明显增加(P<0.05),GABA细胞凋亡数量明显减少(P<0.05)。免疫荧光和Western blot结果显示,与模型组比较,随着加巴喷丁浓度的增加,Bax的阳性表达率明显降低,Bcl-2和CACNA2D1的阳性表达率明显增高。qPCR检测Bax、Bcl-2和CACNA2D1 mRNA表达水平与免疫荧光和Western blot结果一致。结论加巴喷丁能够上调靶蛋白CACNA2D1的蛋白表达,抑制促凋亡蛋白Bax,促进凋亡抑制因子Bcl-2的表达。Aim To identify the molecular target of gabapentin in the treatment of postherpetic neuralgia(PHN).Methods The molecular target of gabapentin for PHN was analyzed by network pharmacology and molecular docking and confirmed by coprecipitation test.Rats were randomly divided into control group,model group,model+50 mg·kg^(-1) gabapentin group,model+100 mg·kg^(-1) gabapentin group,and model+200 mg·kg^(-1) gabapentin group,with nine rats in each group.The pain-related behaviors of the rats were measured at different time points.The mRNA and protein expressions of CACNA2D1,Bax,and Bcl-2 in rat spinal cord were determined by immunofluorescence,Western blot,and qPCR.Results CACNA2D1 was the target gene of gabapentin that determined via network pharmacology,molecular docking,and co-precipitation tests.After modeling,mechanical pain threshold and thermal pain threshold significantly decreased,and the number of apoptotic GABA cells significantly increased.However,after intraperitoneal injection of 50,100,and 200 mg·kg^(-1) gabapentin,mechanical pain threshold and thermal pain threshold significantly increased(P<0.05),and the number of apoptotic GABA cells significantly decreased(P<0.01).Immunofluorescence and Western blot results showed that compared with the model group,with the increase of gabapentin concentration,the positive expression rate of Bax significantly decreased,and the positive expression rate of Bcl-2 and CACNA2D1 significantly increased.The mRNA expression levels of Bax,Bcl-2 and CACNA2D1 detected by qPCR were consistent with the results of immunofluorescence and Western blot.Conclusions Gabapentin up-regulates the expression of target protein CACNA2D1,inhibits the proapoptotic protein Bax,and promotes the expression of apoptotic inhibitor Bcl-2.

关 键 词：加巴喷丁 网络药理学 带状疱疹后神经痛 疼痛行为学 细胞凋亡 CACNA2D1 BAX BCL-2 

分 类 号：R-332[医药卫生] R319R329.25R441.1R752.1R745R971.6