臭氧抑制慢性压迫性坐骨神经损伤模型小鼠的脊髓Homer1b/c对神经病理性疼痛的影响  

Inhibitory effect of ozone on neuropathic pain in the spinal cord Homer1b/c of mice with chronic constriction injury of the sciatic nerve

作　　者：王前伦 钱怡玲 王志萍 Wang Qianlun;Qian Yiling;Wang Zhiping(Department of Anesthesiology and Critical Care Medicine,Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medi-cine,Shanghai 200082,China;Department of Anesthesiology,Wuxi People's Hospital Affiliated to Nanjing Medical University,Wuxi 214043,China;Affiliated Hospital of Xuzhou Medical University,Xuzhou 221002,China)

机构地区：[1]上海交通大学医学院附属新华医院麻醉与重症医学科,上海200082 [2]南京医科大学附属无锡人民医院麻醉科,无锡214043 [3]徐州医科大学附属医院,徐州221002

出　　处：《国际麻醉学与复苏杂志》2023年第10期1038-1044,共7页International Journal of Anesthesiology and Resuscitation

摘　　要：目的研究臭氧对慢性压迫性坐骨神经损伤(chronic constriction injury of the sciatic nerve,CCI)模型小鼠疼痛的影响,探讨CCI模型小鼠脊髓Homer1b/c在此过程中的作用。方法按随机数字表法将62只雄性昆明小鼠分为8组(每组8只):假手术组(S1组)、注射纯氧对照组(S2组)、CCI术后第7天组(C1组)、CCI术后第13天组(C2组)、术后第7天注射臭氧组(O1组)、术后第7~13天注射臭氧组(O2组)、鞘内注射反义寡核苷酸组(AS组)、鞘内注射生理盐水对照组(NS组)。各组小鼠均选择左侧坐骨神经建立CCI模型,术后7 d小鼠出现左后肢负重减少、挛缩等体征,且无肢体瘫痪、自噬等现象视为建立模型成功。术后第7天开始观测机械缩足阈值(mechanical withdrawal threshold,MWT)和热缩足潜伏期(thermal withdraw latency,TWL),Western blot法检测脊髓Homer1b/c和代谢型谷氨酸受体(metabotropic glutamate receptor,mGluR)5表达水平,免疫荧光法检测Homer1b/c和mGluR5在脊髓的分布与表达。S1组只游离神经不结扎,术后第7天观测MWT和TWL;S2组行左侧坐骨神经结扎,逐层缝合,在术后第7天注射纯氧2、4、8、24 h后观测MWT;C1组行左侧坐骨神经结扎,逐层缝合,术后第7天观测MWT和TWL,其中MWT观测时间点同O1组;C2组行左侧坐骨神经结扎,逐层缝合,术后第7~13天观测MWT和TWL;O1组行左侧坐骨神经结扎,逐层缝合,在术后第7天注射臭氧,观测注射后2、4、8、24 h的MWT和注射后2 h的TWL;O2组行左侧坐骨神经结扎,逐层缝合,在术后第7~13天每天注射臭氧2 h后观测MWT;AS组行左侧坐骨神经结扎,逐层缝合,在术后第4天起鞘内注射Homer1b/c的反义寡核苷酸至第7天,术后第4~9天观测MWT;NS组为AS组的生理盐水对照组,与AS组相同时间点观测MWT。结果与S1组比较,C1和C2组小鼠MWT及TWL均降低(P<0.05);与C1组比较,O1组小鼠MWT升高(P<0.05),而TWL差异无统计学意义(P>0.05);与C1组比较,O1组小鼠MWT在注射臭氧2、4 h后�Objective To investigate the effect of ozone on the pain in mice with chronic constriction injury of the sciatic nerve(CCI)and discuss the role of Homer1b/c in the spinal cord of CCI mice during the process.Methods According to the random number table method,64 male Kunming mice were divided into eight groups(n=8):a sham surgery group(group S1),a pure oxygen in‑jection control group(group S2),a 7‑day after CCI treatment group(group C1),a 13‑day after CCI treatment group(group C2),an ozone injection group on post‑treatment 7 d(group O1),an ozone injection group on post‑treatment 7−13 d(group O2),an antisense oligonu‑cleotide injection group(group AS),and an intrathecal injection of normal saline control group(group NS).In the current study,the left sciatic nerve was selected to establish the CCI model and those showing the signs,such as reduced weight bearing and contracture of the left hind limb 7 d after treatment,without limb paralysis and autophagy were considered as a success model.On post‑treatment day 7,the mechanical withdrawal threshold(MWT)and thermal withdrawal latency(TWL)were recorded.The levels of spinal cord Hom‑er1b/c and metabotropic glutamate receptor(mGluR)5 were detected by Western blot.The distribution and expression of Homer1b/c and mGluR5 in the spinal cord were determined by immunofluorescence.In group S1,the sciatic nerve was exposed but not ligated,and the MWT and TWL were recorded on post‑treatment day 7.In group S2,the left sciatic nerve was ligated,sutured layer by layer,and the MWT was recorded after injection of pure oxygen on post‑treatment 2,4,8 h and 24 h.In group C1,the left sciatic nerve was li‑gated,sutured layer by layer,and the MWT and TWL were recorded on post‑treatment day 7,where the MWT was observed on the same time point of group O1.In group C2,the left sciatic nerve was ligated,sutured layer by layer,and the MWT and TWL were ob‑served 7−13 d after treatment.In group O1,the left sciatic nerve was ligated,sutured layer by layer,and ozone was

关 键 词：神经病理性疼痛 臭氧 Homer1b/c 代谢型谷氨酸受体5 

分 类 号：R-332[医药卫生] R688