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自噬在神经病理性痛大鼠脊髓星形胶质细胞活化中的作用

Role of autophagy in activation of astrocytes in spinal cord of rats with neuropathic pain

摘要:

目的 评价自噬在神经病理性痛大鼠脊髓星形胶质细胞活化中的作用.方法 雄性SD大鼠136只,8~10周,体重250~ 300 g,采用随机数字表法分为4组(n=34):假手术组(S组)、神经病理性痛组(NP组)、自噬抑制剂3-甲基腺嘌呤组(MA组)和自噬激动剂雷帕霉素组(R组).采用慢性压迫性损伤法制备大鼠神经病理性痛模型.MA组和R组于造模前1h时分别腹腔注射3-甲基腺嘌呤15 mg/kg和雷帕霉素10 mg/kg.随机取10只大鼠,分别于造模前和造模后1、3、7、14 d时,测定机械缩足反应阈(MWT)和热缩足潜伏期(TWL).分别于造模前和造模后1、3、7d时,随机处死6只大鼠,取L4-6脊髓组织,采用Western blot法检测自噬蛋白微管相关蛋白1轻链3Ⅱ(LC3Ⅱ)、Beclin-1和胶质纤维酸性蛋白(GFAP)的表达.结果 与S组比较,NP组造模后各时点MWT降低,TWL缩短,脊髓LC3Ⅱ、Beclin-1和GFAP的表达上调(P<0.05);与NP组比较,MA组造模后各时点MWT降低,TWL缩短,脊髓LC3Ⅱ和Beclin-1的表达下调,GFAP表达上调,R组造模后各时点MWT升高,TWL延长,脊髓LC3Ⅱ和Beclin-1的表达上调,GFAP表达下调(P<0.05).结论 自噬可通过促进脊髓星形胶质细胞活化,参与大鼠神经病理性痛的形成和维持.

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Objective To evaluate the role of autophagy in activation of astrocytes in the spinal cord of rats with neuropathic pain (NP).Methods One hundred thirty-six male Sprague-Dawley rats, aged 8-10 weeks, weighing 250-300 g, were randomly divided into 4 groups (n =34 each) using a random number table: sham operation group (group S), group NP, autophagy inhibitor 3-methyladenine (3-MA) group (group MA), and autophagy activator rapamycin group (group R).NP was induced by chronic constriction injury.In 3-MA and R groups, 3-MA 15 mg/kg and rapacymin 10 mg/kg were injected intraperitoneally, respectively, at 1 h before NP.Ten rats were randomly selected, and the mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured before NP and at 1, 3, 7 and 14 days after NP.Before NP and at 1, 3 and 7 days after NP, 6 rats were randomly sacrificed, the L4-6 segments of the spinal cord was harvested to detect microtubule-associated protein 1 light chain 3 Ⅱ (LC3 Ⅱ), Beclin-1 and glial fibrillary acidic protein (GFAP) expression by Western blot.Results Compared with group S, the MWT was significantly decreased, the TWL was shortened, and the expression of LC3 Ⅱ , Beclin-1 and GFAP was up-regulated at each time point after NP in group NP (P< 0.05).Compared with group NP, the MWT was significantly decreased, the TWL was shortened, the expression of LC3 Ⅱ and Beclin-1 was down-regulated, and the expression of GFAP was up-regulated at each time point after NP in group MA, and the MWT was significantly increased, the TWL was prolonged, the expression of LC3 Ⅱ and Beclin-1 was up-regulated, and the expression of GFAP was down-regulated at each time point after NP in group R (P<0.05).Conclusion Autophagy is involved in the development and maintenance of NP through promoting the activation of astrocytes in the spinal cord of rats.

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作者: 陈红光 [1] 陈亚军 [1] 谢克亮 [1] 于泳浩 [1]
期刊: 《中华麻醉学杂志》2015年35卷11期 1307-1309页 ISTICPKUCSCD
栏目名称: 疼痛诊疗与研究
DOI: 10.3760/cma.j.issn.0254-1416.2015.11.005
发布时间: 2016-01-08
基金项目:
国家自然科学基金 天津市自然科学基金
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