GM-1预处理对布比卡因脊神经毒性模型大鼠脊髓组织JNK信号通路的影响  

Influence of GM-1 preconditioning on JNK signal pathway in bupivacaine-induced spinal toxicity rats

作　　者：罗茜 刘本铨 吉杰梅 封青 刘敬臣 LUO Xi;LIU Benquan;JI Jiemei;FENG Qing;LIU Jingchen(The First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学第一附属医院

出　　处：《山东医药》2019年第23期27-32,共6页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81660623)

摘　　要：目的评价单唾液酸四己糖神经节苷脂(GM-1)预处理对布比卡因神经毒性模型大鼠脊髓组织c-Jun氨基末端激酶(JNK)信号通路的影响。方法选择108只SPF级雄性SD大鼠随机均分为三组:生理盐水组(saline组)、布比卡因组(S+B组)和GM-1预处理组(G+B组)。三组大鼠行鞘内置管后分别以下处理:G+B组鞘内注射30 mg/kg GM-1作为布比卡因注射前的预处理;saline组和S+B组同时鞘内注射等量生理盐水。24 h后S+B组、G+B组重复鞘内注射5%布比卡因0.12μL/g共3次,每次间隔1.5 h,建立布比卡因脊髓神经毒性模型;saline组同时鞘内注射等量生理盐水。各组于建模后0、1、3、5、7、14 d测大鼠甩尾反应潜伏期(TFL),换算成最大抗辐射热效应百分比(%MPE)以进行感觉功能的评估,并行运动功能BBB评分。随后于每个时点随机处死6只大鼠,取腰膨大处脊髓组织,光镜、电镜下观察并进行组织损伤评分(SD评分),TUNEL法检测细胞凋亡率,Western blotting法检测脊髓组织p-JNK蛋白表达,qRT-PCR法检测脊髓组织JNK mRNA表达。结果与saline组比较,S+B组和G+B组在注药后各时点%MPE、SD评分、细胞凋亡率、JNK mRNA的相对表达量、p-JNK蛋白相对表达量均升高,BBB评分降低(P均<0.05);与S+B组比较,G+B组%MPE在注药后3、5、7、14 d降低,BBB评分在5、7、14 d升高,SD评分、细胞凋亡率在5、7、14 d降低,JNK mRNA在3、5、7 d降低,p-JNK相对表达量在5、7 d降低(P均<0.05)。结论GM-1预处理可以减轻大鼠鞘内注入布比卡因引起的脊神经毒性,改善大鼠的感觉、运动功能,减轻神经细胞凋亡和病变,其机制与抑制JNK信号通路的激活有关。Objective To evaluate the influence of monosialoganglioside(GM-1)preconditioning on c-Jun N-terminal kinase(JNK)signal pathway in bupivacaine-induced spinal toxicity rats.Methods One hundred and eight adult male Sprague-Dawley rats were randomly divided into 3 groups(n=36 each):the saline group,saline and bupivacaine group(group S+B),and GM-1 and bupivacaine group(group G+B).Every rat from the three groups was implanted an intrathecal catheter.Rats in the group G+B were injected with GM-1 at a dose of 30 mg/kg via the implanted intrathecal catheter 24 h before bupivacaine injection as an pretreatment,while rats in the group S+B and group saline received equal volume of normal saline at the same time.Neurotoxicity models were prepared by injecting 5%bupivacaine at a dose of 0.12μL/g via the intrathecal catheter at 90-minute intervals for 4.5 h in the group S+B and group G+B,while the saline group received equal volume of normal saline.The recovery of the locomotor function was evaluated by Basso,Beattie and Bresnahan(BBB)score and sensory function by tail-flick latency(TFL)at 0,1,3,5,7,and 14 d after bupivacaine injection.TFL was converted to the percent maximum possible effect(%MPE).Six rats were sacrificed in each group at each time point,and lumbosacral enlargement was taken for the further examination.Light and electron microscopy were applied in observing,evaluating and scoring the histological injury in lumbosacral enlargement.We evaluated the apoptosis rate of spinal cells by TUNEL.JNK mRNA and p-JNK protein expression levels were detected by qRT-PCR and Western blotting,respectively.Results Compared with the saline group,%MPE,histological injury score,apoptosis rate,JNK mRNA and p-JNK protein expression were significantly higher,and BBB scores were significantly lower in the groups S+B and group G+B than in the saline group at all the time points after bupivacaine injection(all P<0.05).Compared with the group S+B,group G+B had lower%MPE at 3,5,7 and 14 d,lower histological injury score and apoptosis rates

关 键 词：布比卡因 神经毒性 单唾液酸四己糖神经节苷脂 JNK信号通路 大鼠 

分 类 号：R614.2[医药卫生—麻醉学]