摘要
目的:探讨脊髓中活性氧(reactive oxygen species,ROS)是否通过激活自噬参与大鼠2型糖尿病神经病理性疼痛(diabetic neuropathic pain,DNP)的形成和发展。方法:清洁级雄性SD大鼠高脂高糖喂养8周,后链脲佐菌素(streptozocin,STZ)单次小剂量(35 mg/kg)腹腔注射,注射后3 d血糖≥16.7 mmol/L为成功诱导2型糖尿病发生;注射后14 d行为学测定痛阈下降至基础值的85%以下,即为成功制备出大鼠2型DNP模型,否则为2型糖尿病不痛组(PL组)。将造模成功的2型DNP大鼠随机分为3组,每组10只:DNP组、DNP+ROS清除剂组(DNP+PBN组)和溶剂对照组(SC组)。另取10只正常SD大鼠为对照组(C组),普通饲料喂养,给予PBN后3 d、7 d、14 d时测定所有组别大鼠的机械缩足反应阈值(mechanical withdrawal threshold,MWT)和热缩足潜伏期(thermal withdrawal latency,TWL),随后处死大鼠,取脊髓腰膨大L4-L6,用免疫印迹法测定脊髓中Beclin 1、LC3II/LC3I和p62的表达水平;用流式细胞仪测定脊髓组织中活性氧(ROS)的含量。结果:与C组和PL组比较,DNP组和SC组于模型制备成功后3 d、7 d和14 d时MWT下降,TWL缩短,脊髓ROS、Beclin 1和LC3II/LC3I表达上调(P<0.05),而p62表达下降(P<0.05);与DNP组比较,DNP+PBN组在3 d、7 d和14 d时MWT升高、TWL延长,脊髓Beclin 1和LC3II/LC3I表达下降(P<0.05),而p62表达上调(P<0.05);DNP组与SC组各指标比较差异均无统计学意义。结论:脊髓ROS通过激活自噬参与调节2型糖尿病神经病理性疼痛的产生和维持。
Objective:To explore whether ROS modulates the initiation,development and maintenance of type 2 diabetic neuropathic pain(DNP)in rat via activating autophagy.Methods:Pathogen-free male Sprague-Dawley rats were fed with a high-fat and high-sugar diet for 8 weeks,and then received a single intraperitoneal streptozocin(STZ)injection with a dose of 35 mg/kg.Three days after the STZ injection,rats with blood glucose level of 16.7 mmol/L or higher were considered type 2 diabetes mellitus.Diabetic rats with neuropathic pain were defined as both mechanical withdrawal threshold(MWT)and thermal withdrawal latency(TWL)≤85%of base value on 14d after STZ injection,while both TWL and MWT>85%of base value was defined as PL group.The type 2 DNP rats were randomly divided into 3 groups(n=10):DNP group,DNP+PBN group(DNP rats treated with ROS scavenger PBN)and SC group(solvent control group).Another 10 normal SD rats were selected as control group(C group)and were fed with common diet.MWT and TWL were measured at 3d,7d and 14d after dosing.The lumbar segments 4-6 of the spinal cord were removed at the same time for determination of the expression of Beclin 1,LC3II/LC3I,and p62 by Western blot;the production of ROS was measured by flow cytometry.Results:Compared with C and PL groups,MWT was significantly decreased,TWL was shortened,ROS production and the expression of Beclin 1 and LC3II/LC3I in the spinal cord were up-regulated at 3,7 and 14 d in DNP and SC groups(P<0.05);while the expression of p62 was decreased(P<0.05).Compared with DNP group,MWT was significantly increased,TWL was prolonged,the expression of Beclin 1 and LC3II/LC3I in spinal cord was decreased and p62 was increased at 3,7 and 14 d in DNP+PBN group(P<0.05).There is no significant difference in all measurment between DNP group and SC group(P>0.05).Conclusion:ROS modulates the initiation,development and maintenance of type 2 diabetic neuropathic pain(DNP)via activating autophagy.
作者
陈佳丽
张茂表
陆嘉辉
贾改丽
李军
曹红
CHEN Jia-Li;ZHANG Mao-Biao;LU Jia-Hui;JIA Gai-Li;LI Jun;CAO Hong(Department of Anesthesiology,Second Affiliated Hospital of Wenzhou Medical University,Pain Medicine Institute of Wenzhou Medical University,Wenzhou 325027,China)
出处
《中国疼痛医学杂志》
CAS
CSCD
北大核心
2020年第1期27-34,共8页
Chinese Journal of Pain Medicine
基金
国家自然科学基金（81771487）
浙江省自然科学基金（LY17H070006）
关键词
2型糖尿病
神经病理性疼痛
活性氧
自噬
脊髓
Neuropathic pain
Type 2 diabetes mellitus
Reactive oxygen species
Autophagy
Spinal cord
