罗西格列酮对脊髓损伤大鼠后肢运动功能恢复的影响  

Effect of rosiglitazone on motor function recovery of hind limbs in rats with spinal cord injury

作　　者：胡莉琼 奉振成 张延祠 崔树廷[4] 王敏[2] 孟庆奇 Hu Liqiong;Feng Zhencheng;Zhang Yanci;Cui Shuting;Wang Min;Meng Qingqi(Department of Intensive Care Unit,Guangzhou Red Cross Hospital,Jinan University Medical College,Guangzhou 510220,China;Department of Orthopedics,Guangzhou Red Cross Hospital,Jinan University Medical College,Guangzhou 510220,China;Department of Orthopedics,Second Hospital of Shijiazhuang,Shijiazhuang 050000,China;Department of Orthopedics,Second Hospital of Liaocheng Affiliated to Taishan Medical College,Liaocheng 252600,China)

机构地区：[1]暨南大学医学院附属广州红十字会医院重症医学科,510220 [2]暨南大学医学院附属广州市红十字会医院骨科,510220 [3]石家庄市第二人民医院骨科,050000 [4]泰山医学院附属聊城市第二医院骨科,252600

出　　处：《中华创伤杂志》2019年第6期568-576,共9页Chinese Journal of Trauma

基　　金：国家自然科学基金(81301038).

摘　　要：目的探讨过氧化物增殖体激活受体-γ(PPAR-γ)激动剂罗西格列酮对大鼠脊髓损伤后下肢运动恢复的影响。方法选择68只雌性SD大鼠采用改良Allen法制造脊髓损伤模型。(1)8只按照随机数字表法分为阴性对照组和罗西格列酮组,每组4只。采用免疫组化染色检测各组伤后7 d大鼠脊髓中天冬氨酸蛋白水解酶-1(caspase-1)的表达。(2)48只按照随机数字表法分为阴性对照组、罗西格列酮组、罗西格列酮+壳梭孢菌素[核因子-κB(NF-κB)激活剂]组、罗西格列酮+单钠尿酸盐[寡聚化结构域样受体蛋白3(NLRP3)激动剂]组,每组12只。各组伤后1,3,14,21,28 d行后肢运动功能BBB评分;伤后28 d采用ELISA检测各组白细胞介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)的蛋白表达量。(3)12只用于大鼠脊髓小胶质细胞的提取,培养7 d后小胶质细胞随机分为阴性对照组(不加药物处理)、罗西格列酮组(1μmol/L罗西格列酮处理)、罗西格列酮+壳梭孢菌素组(1μmol/L罗西格列酮+20μmol/L壳梭孢菌素处理)、壳梭孢菌素组(20μmol/L壳梭孢菌素处理)和壳梭孢菌素+MCC950(NLRP3拮抗剂)组(20μmol/L壳梭孢菌素+100 nmol/L MCC950)。采用Western blot检测小胶质细胞中caspase-1、NF-κB和NLRP3的蛋白表达量。采用ELISA检测小胶质细胞培养的上清液中IL-1β和TNF-α的蛋白表达量。结果与阴性对照组比较,罗西格列酮组脊髓损伤区域caspase-1表达减少[灰质区:(5.1±0.8)∶(6.9±1.1),白质区:(5.6±0.9)∶(7.5±1.1)](P<0.05)。伤后28 d,罗西格列酮组BBB评分[(14.7±1.6)分]效果最佳,罗西格列酮+壳梭孢菌素组为(10.5±2.1)分,优于罗西格列酮+单钠尿酸组的(7.2±1.3)分(P<0.05)。伤后28 d,罗西格列酮组IL-1β和TNF-α的表达低于其他各组(P<0.05)。在体外小胶质细胞中,罗西格列酮组caspase-1、NF-κB、IL-1β及TNF-α的表达量均低于阴性对照组(P<0.05);罗西格列酮+壳梭孢菌素组caspase-1、NLRP3、IL-1β及TNF-Objective To investigate the effect of the peroxide proliferator-activated receptor-gamma(PPAR-γ)agonist rosiglitazone on the motor function recovery of hind limbs in rats with spinal cord injury.Methods Sixty-eight female SD rats were used to establish spinal cord injury model by modified Allen method.(1)Eight rats were randomly divided into negative control group and rosiglitazone group with four rats in each group.The expression of aspartate proteolytic enzyme-1(caspase-1)in spinal cord of rats 7 days after injury was detected by immunohistochemical staining.(2)Forty-eight rats were randomly divided into negative control group,rosiglitazone group,rosiglitazone+Clostridium chitosans group[nuclear factor kappa B(NF-kappa B)activator],rosiglitazone+monosodium urate group[oligomerization domain-like receptor protein 3(NLRP3)antagonist],with 12 rats in each group.BBB scores of hindlimb motor function were assessed at 1,3,14,21 and 28 days after injury in each group.The expression of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)in each group was detected by ELISA at 28 days after injury.Microglia were isolated from the spinal cord of 12 rats and cultured for 7 days.They were randomly divided into the following five groups:(1)negative control group:no drug treatment;(2)rosiglitazone group:1 micromol/L rosiglitazone treatment;(3)rosiglitazone+Clostridium chitin group:1 micromol/L rosiglitazone+20 micromol/L Clostridium chitosporin treatment;(4)Clostridium chitosan treatment Mycoplasma group:20μmol/L shell Clostridium treatment;(5)Clostridium chitosanin+MCC950 group[(NLRP3)antagonist]:20μmol/L Clostridium chitosanin+100 nmol/L MCC950;Western blot was used to detect the expressions of caspase-1,NF-kappa B and NLRP3 in microglia cells;ELISA was used to detect the expressions of IL-1βand TNF-αin the supernatant of microglia culture.Results Compared with negative control group,caspase-1 expression was decreased in rosiglitazone group in spinal cord injury area[gray matter area:5.1±0.8∶6.9±1.1;white m

关 键 词：脊髓损伤 罗西格列酮 小神经胶质细胞 大鼠 

分 类 号：R651.2[医药卫生—外科学]