双氢青蒿素联合七氟醚调控PERK/CHOP通路对乙型脑炎病毒NS1蛋白诱导的小鼠神经病理性疼痛的影响  

Effects of Dihydroartemisinin Combined with Sevoflurane on Neuropathic Pain Induced by the NS1 Protein of Japanese Encephalitis Virus via Regulation of the PERK/CHOP Pathway

作　　者：罗琼[1] 彭思思[1] 罗晶[1] LUO Qiong;PENG Sisi;LUO Jing(Zhongnan Hospital of Wuhan University,Wuhan 430071,China)

机构地区：[1]武汉大学中南医院,武汉430071

出　　处：《病毒学报》2024年第6期1274-1280,共7页Chinese Journal of Virology

摘　　要：乙型脑炎病毒(Japanese encephalitis virus,JEV)是病毒性脑炎的主要病因,其感染引起神经病理性疼痛的致病机制目前尚不明确。因此,迫切需要阐明其作用机制。治疗神经性疼痛的药物由于其不良反应多、疗效不佳等原因,寻找新的治疗药物显得尤为必要。本研究首先使用JEV感染小鼠,观察脑组织病理学变化、分析病原分布;检测机械缩足反射阈值(Mechanical withdrawal threshold,MWT)和热缩足反射潜伏期(Thermal withdrawal latency,TWL);检测大脑中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和丙二醛(MDA)含量;Western blot实验探究了NS1、PERK、CHOP相关通路蛋白表达。随后我们选用了双氢青蒿素联合七氟醚的治疗方案,并对其疗效和作用机制进行了探究。实验结果显示,JEV感染后,荧光标记病毒周围大量小胶质细胞活化、神经元细胞坏死,小鼠MWT和TWL明显下降,脑组织出现明显病理损伤,SOD和CAT水平降低,MDA水平升高,NS1、PERK、CHOP表达明显上调。双氢青蒿素联合七氟醚治疗后,小鼠脑组织病理损伤显著缓解,且氧化损伤被抑制,小鼠MWT和TWL明显上升,NS1、PERK、CHOP表达明显下调。因此我们得出结论,双氢青蒿素联合七氟醚可通过PERK/CHOP通路抑制神经元细胞氧化应激损伤,进而发挥其神经病理性疼痛保护作用。本研究阐述了JEV感染机制,同时预估了JEV中的NS1蛋白具有作为神经病理性疼痛治疗靶点的潜在价值。Japanese encephalitis virus(JEV)is a major cause of viral encephalitis,yet the pathogenic mechanisms underlying JEV-induced neuropathic pain remain unclear.Therefore,it is critical to elucidate these mechanisms.Existing treatments for neuropathic pain often result in side effects and limited efficacy,highlighting the need for novel therapeutic approaches.In this study,mice were infected with JEV to observe the pathological changes in brain tissues and to analyze the distribution of viral pathogens.Mechanical withdrawal threshold(MWT)and thermal withdrawal latency(TWL)were assessed,and the levels of super oxide dismutase(SOD),catalase(CAT),and malondialdehyde(MDA)in brain tissue were measured.Westernblot analysis was performed to examine the expression of NS1,PERK,and CHOP proteins in relevant pathways.Subsequently,a treatment regimen combining dihydroartemisinin and sevoflurane was administered,and its therapeutic efficacy and underlying mechanisms were explored.Experimental results showed that JEV infection caused significant microglia activation,neuronal necrosis,reduced MWT and TWL,severe pathological damage to brain tissue,decreased SOD and CAT levels,increased MDA levels,and elevated expression of NS1,PERK,and CHOP.Treatment with dihydroartemisinin and sevoflurane significantly alleviated brain tissue pathology,reduced oxidative stress,improved MWT and TWL,and downregulated the expression of NS1,PERK,and CHOP.These findings suggest that the dihydroartemisinin combined with sevoflurane mitigates oxidative stress in neurons through PERK/CHOP pathway,thereby exerting a protective effect against neuropathic pain.This study not only elucidates the mechanisms of JEV infection but also identifies the NS1 protein as a potential therapeutic target for JEV-induced neuropathic pain.

关 键 词：双氢青蒿素 七氟醚 PERK/CHOP 乙型脑炎病毒 NS1蛋白 神经病理性疼痛 

分 类 号：R373.3[医药卫生—病原生物学]