激活SIRT1可通过抑制背根神经节神经元线粒体损伤缓解紫杉醇诱导的神经病理性疼痛  

SIRT1 activation alleviates paclitaxel induced neuropathic pain by inhibiting mitochondrial damage in the dorsal root ganglion neurons

作　　者：曾雁雁 林丽 姚孟宇 吴文[5] 黄怀 Zeng Yanyan;Lin Li;Yao Mengyu;Wu Wen;Huang Huai(Department of Hyperbaric Oxygen Medicine and Rehabilitation(Intensive Care Rehabilitation Center),General Hospital of Southern Theater Command,Guangzhou 510010,China;Department of Rehabilitation Medicine,Zhujiang Hospital,Southern Medical University;Graduate School,Guangzhou University of Chinese Medicine,Guangzhou 510006,China;Department of Bone Oncology,Guangdong Provincial People's Hospital(Guangdong Academy of Medical Sciences),Guangzhou 510080,China;Department of Rehabilitation Medicine,Zhujiang Hospital,Southern Medical University,Guangzhou 510282,China)

机构地区：[1]解放军南部战区总医院高压氧康复科(重症康复中心),广州510010 [2]南方医科大学珠江医院康复医学科博士后工作站 [3]广州中医药大学研究生院,广州510006 [4]广东省人民医院(广东省医学科学院)骨肿瘤科,广州510080 [5]南方医科大学珠江医院康复医学科,广州510282

出　　处：《中华神经医学杂志》2024年第10期983-991,共9页Chinese Journal of Neuromedicine

基　　金：国家自然科学基金青年科学基金(82102650);广东省基础与应用基础研究基金(2020A1515110704)。

摘　　要：目的探讨激活沉默信息调节因子1(SIRT1)能否通过抑制背根神经节神经元线粒体损伤缓解紫杉醇诱导的神经病理性疼痛。方法48只健康雄性SD大鼠按随机数字表法分为溶剂对照组、紫杉醇组、紫杉醇+SIRT1抑制剂组、紫杉醇+SIRT1激活剂组,每组12只。后3组大鼠应用腹腔注射紫杉醇方式制备成神经病理性疼痛模型,注射剂量为8 mg/kg,注射时间点分别为实验第1、4、7天;后2组大鼠分别于第1次注射紫杉醇前30 min时单次鞘内注射SIRT1抑制剂EX527或激活剂SRT1720。另再取12只大鼠用相同方法制备成神经病理性疼痛模型(模型组),分别于实验前1 d及实验第3、7、14天时采用Western blotting实验检测大鼠背根神经节组织中SIRT1的表达。实验前1 d及实验第3、7、14天时,利用Von-Frey纤维丝检测溶剂对照组、紫杉醇组、紫杉醇+SIRT1抑制剂组、紫杉醇+SIRT1激活剂组大鼠后足50%机械刺激缩足反射阈值,利用热辐射热痛检测仪评估热刺激缩足反射潜伏期。实验第7天疼痛行为学评估结束后过量麻醉处死各组6只大鼠,快速分离L_(4)~L_(6)背根神经节组织并培养原代神经元,分别采用Western blotting实验检测背根神经节组织中SIRT1的表达,利用JC-1线粒体膜电位检测试剂盒检测线粒体膜电位(以红绿荧光强度比率表示),利用过氧化氢(H_(2)O_(2))检测试剂盒检测H_(2)O_(2)浓度,利用线粒体超氧化物检测试剂盒和线粒体绿色荧光探针试剂盒检测线粒体超氧化物的表达。结果与实验前1 d相比,模型组大鼠实验第3、7、14天背根神经节组织中SIRT1表达均明显降低,差异均有统计学意义(P<0.05)。实验第3、7、14天时,与溶剂对照组相比,紫杉醇组大鼠的50%机械刺激缩足反射阈值[(6.37±2.27)g、(5.47±2.42)g、(5.34±1.74)g]、热刺激缩足反射潜伏期[(9.38±1.27)s、(9.70±1.97)s、(9.12±1.21)s]均明显降低,差异均有统计学意义(P<0.05);与紫杉醇组相�ObjectiveTo investigate whether silencing information regulator 1(SIRT1)activation can relieve paclitaxel-induced neuropathic pain by inhibiting mitochondrial damage in dorsal root ganglion neurons.MethodsForty-eight healthy male SD rats were randomly divided into solvent control group,paclitaxel group,paclitaxel+SIRT1 inhibitor group and paclitaxel+SIRT1 agonist group(n=12).Neuropathic pain model in the later 3 groups was prepared by intraperitoneal injection of paclitaxel at 8 mg/kg on the 1 st,4 th and 7 th d of experiment,respectively;rats in the paclitaxel+SIRT1 inhibitor group and paclitaxel+SIRT1 agonist group were respectively injected with SIRT1 inhibitor EX527 or agonist SRT172030 min before the first injection of paclitaxel.In addition,neuropathic pain model was established in 12 rats(model group)by the same method and SIRT1 expression in the dorsal root ganglion tissues was detected by Western blotting 1 d before experiment and on the 3 rd,7 th and 14 th d of experiment,respectively.Von-Frey filament was used to detect the 50%paw withdrawal mechanical threshold(PWMT),and thermal radiation thermal pain detector was used to evaluate the paw withdraw thermal latency(PWTL)1 d before experiment and on the 3 rd,7 th and 14 th d of experiment.On the 7 th d of experiment,6 rats in each group were sacrificed with excessive anesthesia after PWMT and PWTL detection;L_(4)-L_(6)dorsal root ganglion tissues were rapidly isolated and primary neurons were cultured;Western blotting was used to detect SIRT1 expression in the dorsal root ganglion tissues,JC-1 mitochondrial membrane potential detection kit was used to detect mitochondrial membrane potential(ratio of orange-red fluorescence to green fluorescence),hydrogen peroxide(H_(2)O_(2))detection kit was used to detect H_(2)O_(2)concentration,and mitochondrial superoxide detection kit and mitochondrial green fluorescence probe kit were used to detect mitochondrial superoxide expression.ResultsIn the model group,SIRT1 expression in the dorsal root ganglion tissues one

关 键 词：神经病理性疼痛 沉默信息调节因子1 紫杉醇 线粒体 氧化应激 

分 类 号：R741[医药卫生—神经病学与精神病学]