海马ATF5在神经病理性痛致小鼠认知功能障碍中的作用:与mtUPR的关系  

Role of hippocampal activating transcription factor 5 in cognitive impairment induced by neuropathic pain in mice:relationship with mitochondrial unfolded protein response

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作  者:邢飞 石小山 许耀威 卫新[1] 渠明翠 程丹[1] 袁静静[1] 王中玉[1] 邢娜[1] 李艳娜[1] Xing Fei;Shi Xiaoshan;Xu Yaowei;Wei Xin;Qu Mingcui;Cheng Dan;Yuan Jingjing;Wang Zhongyu;Xing Na;Li Yanna(Department of Anesthesiology,Pain and Perioperative Medicine,The First Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China)

机构地区:[1]郑州大学第一附属医院麻醉与围手术期及疼痛医学部,郑州450000

出  处:《中华麻醉学杂志》2025年第3期329-334,共6页Chinese Journal of Anesthesiology

基  金:国家自然科学基金(8237251082001187);河南省医学科技攻关计划(SBGJ202403023,SBGJ202402050);河南省高等学校重点科研项目计划(24A320071)。

摘  要:目的:评价海马活化转录因子5(ATF5)在神经病理性痛致小鼠认知功能障碍中的作用及其与线粒体未折叠蛋白反应(mtUPR)的关系。方法:实验一:SPF级健康雄性C57BL/6小鼠24只,6~8周龄,体质量20~25 g,采用随机数字表法分为2组(n=12):假手术组(S1组)和神经病理性痛组(NP组)。采用慢性坐骨神经压迫损伤术建立小鼠神经病理性痛模型。于建模前、建模后7、14、21和28 d时测定机械缩足反应阈(MWT)和热缩足潜伏期(TWL),于建模后30~31 d时行新物体识别实验评估小鼠认知功能。新物体识别实验结束后处死小鼠取海马CA1区,采用蛋白免疫印迹法检测ATF5的表达,采用免疫荧光双染技术检测ATF5分别在神经元、小胶质细胞和星形胶质细胞的表达。实验二:SPF级健康雄性C57BL/6小鼠36只,6~8周龄,体质量20~25 g,采用随机数字表法分为3组(n=12):假手术组(S2组)、神经病理性痛+ATF5上调组(NA组)和神经病理性痛+空载病毒组(NE组)。造模后14 d时,NA组和NE组分别于小鼠海马CA1区注射神经元中特异性上调ATF5表达病毒和空载病毒。于造模前、造模后28和35 d时测定MWT和TWL,于造模后36 d时行新物体识别实验评估小鼠认知功能。行为学实验结束后处死小鼠取海马CA1区,采用蛋白免疫印迹法检测ATF5和mtUPR标记蛋白[LON蛋白水解酶1(LONP1)和伴侣蛋白60(HSP60)]的表达。结果:实验一:与S1组相比,NP组造模前MWT和TWL差异无统计学意义(P>0.05),造模后7、14、21和28 d时MWT和TWL降低,造模后31 d时新物体识别指数(DI)降低,海马ATF5表达下调,神经元ATF5表达下调(P<0.05),小胶质细胞和星形胶质细胞ATF5表达差异无统计学意义(P>0.05)。实验二:与S2组相比,NE组和NA组造模后28和35 d时MWT和TWL降低,NE组DI降低,海马ATF5、LONP1和HSP60表达下调(P<0.05),NA组差异无统计学意义(P>0.05);与NE组相比,NA组MWT和TWL差异无统计学意义(P>0.05),DI升高,海马ATF5、LONP1和HSP60表达上Objective:To evaluate the role of hippocampal activating transcription factor 5(ATF5)in cognitive impairment induced by neuropathic pain and the relationship with mitochondrial unfolded protein response(mtUPR)in mice.Methods:This study was conducted in 2 parts.ExperimentⅠTwenty-four SPF healthy male C57BL/6 mice,aged 6-8 weeks,weighing 20-25 g,were divided into 2 groups(n=12 each)using a random number table method:sham operation group(S1 group)and neuropathic pain group(NP group).Neuropathic pain was induced by chronic constriction injury to the sciatic nerve.The mechanical paw withdrawal threshold(MWT)and thermal paw withdrawal latency(TWL)were measured before developing the model and at 7,14,21 and 28 days after developing the model.Mouse cognitive function was assessed using the novel object recognition test from 30-31 days after developing the model.After the end of the novel object recognition test,mice were sacrificed and the hippocampal CA1 region was harvested for determination of the expression of ATF5(by Western blot)and the expression of ATF5 in neurons,microglia and astrocytes(by immunofluorescence double staining).ExperimentⅡThirty-six SPF healthy male C57BL/6 mice,aged 6-8 weeks,weighing 20-25 g,were divided into 3 groups(n=12 each)using a random number table method:sham operation group(S2 group),neuropathic pain+ATF5 up-regulation group(NA group),and neuropathic pain+empty virus group(NE group).On day 14 after developing the model,a virus that specifically up-regulated ATF5 expression in neurons and empty virus were injected into the hippocampal CA1 region.The MWT and TWL were measured at days 28 and 35 after developing the model.The novel object recognition test was performed on day 36 after developing the model to evaluate the cognitive function.After the end of the behavioral test,mice were sacrificed and the hippocampal CA1 region was harvested for detection of the expression of ATF5 and mtUPR marker proteins(Lon protease[LONP1]and heat shock protein 60[HSP60])by Western blot.Results:Experi

关 键 词:转录因子 海马 神经痛 认知功能障碍 线粒体 未折叠蛋白反应 

分 类 号:R741[医药卫生—神经病学与精神病学]

 

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