瑞芬太尼诱发切口痛大鼠痛觉过敏时自噬与氧化应激的关系  

Relationship between autophagy and oxidative stress during remifentanil-induced hyperalgesia in rats with incisional pain

作　　者：顾喜燕 秦超 张海芳 于洋[1] 陈亚军[1] 于泳浩[1] Gu Xiyan;Qin Chao;Zhang Haifang;Yu Yang;Chen Yajun;Yu Yonghao(Department of Anesthesiology,General Hospital of Tianjin Medical University Tianjin Research Institute of Anesthesiology,Tianjin 300052,China)

机构地区：[1]天津医科大学总医院麻醉科天津市麻醉学研究所,天津300052

出　　处：《中华麻醉学杂志》2022年第7期841-844,共4页Chinese Journal of Anesthesiology

摘　　要：目的评价瑞芬太尼诱发切口痛大鼠痛觉过敏时自噬与氧化应激的关系。方法清洁级健康雄性SD大鼠32只,体重230~250 g,2~3月龄,采用随机数字表法分为4组(n=8):切口痛组(I组)、瑞芬太尼+切口痛组(RI组)、自噬抑制剂三甲基腺嘌呤+瑞芬太尼+切口痛组(MRI组)和自噬激活剂雷帕霉素组+瑞芬太尼+切口痛组(RRI组)。I组建立大鼠切口痛模型的同时静脉输注等容量生理盐水60 min,RI组、MRI组和RRI组建立大鼠切口痛模型的同时以1μg·kg^(-1)min^(-1)的速率静脉输注瑞芬太尼60 min;MRI组于造模前12 h时腹腔注射三甲基腺嘌呤15 mg/kg,RRI组于造模前12 h时腹腔注射雷帕霉素10 mg/kg。分别于输注瑞芬太尼或生理盐水前24 h(T0)、输注结束后2、6、24和48 h(T1~4)时测定机械缩足反应阈(MWT)及热缩足潜伏期(TWL)。行为学测试结束后处死大鼠并取脊髓腰膨大节段,采用Western blot法测定自噬蛋白微管相关蛋白1轻链3Ⅱ(LC3Ⅱ)、Beclin-1和P62的表达,采用黄嘌呤氧化酶法测定SOD活性,硫代巴比妥法测定MDA含量。结果与T0时比较,4组大鼠T1~4时PWT降低,TWL缩短(P<0.05);与I组比较,RI组T1~4时MWT降低,TWL缩短,脊髓LC3Ⅱ和Beclin-1表达上调,P62表达下调,SOD活性降低,MDA含量升高(P<0.05);与RI组比较,MRI组T1~4时MWT降低,TWL缩短,脊髓LC3Ⅱ和Beclin-1表达下调,P62表达上调,SOD活性降低,MDA含量升高(P<0.05),RRI组T1~4时MWT升高,TWL延长,脊髓LC3Ⅱ和Beclin-1表达上调,P62表达下调,SOD活性升高,MDA含量降低(P<0.05)。结论自噬通过调节氧化应激水平参与瑞芬太尼诱发切口痛大鼠痛觉过敏的过程。Objective To evaluate the relationship between autophagy and oxidative stress during remifentanil-induced hyperalgesia in the rats with incisional pain.Methods Thirty-two clean-grade healthy male Sprague-Dawley rats,weighing 230-250 g,aged 2-3 months,were divided into 4 groups(n=8 each)using a random number table method:incisional pain group(I group),remifentanil+incisional pain group(RI group),autophagy inhibitor 3-methyladenine+remifentanil+incisional pain group(MRI group)and autophagy agonist rapamycin group+remifentanil+incisional pain group(RRI group).The model of incision pain was developed and the equal volume of normal saline was intravenously infused simultaneously for 60 min.In RI,MRI and RRI groups,the model of incision pain was developed and remifentanil 1μg·kg^(-1)·min^(-1)was simultaneously infused for 60 min.In MRI group,3-methyladenine 15 mg/kg was intraperitoneally injected at 12 h before developing the model,and rapamycin 10 mg/kg was intraperitoneally injected at 12 h before developing the model in RRI group.Thermal paw withdrawal latency(TWL)and mechanical paw withdrawal threshold(MWT)were measured at 24 h before infusion of remifentanil or normal saline(T0)and at 2,6,24 and 48 h after the end of infusion(T1-4).The rats were sacrificed after the end of behavioral testing,and the lumbar enlargement segments of the spinal cord were harvested for determination of the expression of autophagy microtubule-associated protein 1 light chain 3Ⅱ(LC3Ⅱ),Beclin-1 and P62(by Western blot),activity of superoxide dismutase(SOD)(by xanthine oxidase method)and content of malondialdehyde(MDA)(by thiobarbital method).Results Compared with the baseline at T0,PWT was significantly decreased and TWL was shortened at T1-4 in the four groups(P<0.05).Compared with I group,MWT was significantly decreased and TWL was shortened at T1-4,the expression of LC3Ⅱand Beclin-1 was up-regulated,the expression of P62 was down-regulated,SOD activity was decreased,and MDA content was increased in RI group(P<0.05).Compared wi

关 键 词：哌啶类 疼痛 手术后 痛觉过敏 自噬 氧化性应激 

分 类 号：R614[医药卫生—麻醉学]