CAJ | 학술논문

目的评价自噬在大鼠神经病理性痛形成中的作用.方法雄性SD大鼠30只,体重200 ～ 220 g,采用随机数字表法分为3组(n=10):假手术组(sham组)、神经病理性痛组(NP组)和自噬诱导剂雷帕霉素组(Rap组).行L4.5鞘内置管后,NP组和Rap组行左侧L5脊神经结扎术制备大鼠神经病理性痛模型,sham组大鼠仅暴露左侧L5神经但不结扎.Rap组大鼠分别于结扎前30 min和术后2d鞘内注射雷帕霉素60 μg,sham组和NP组注射等容溶剂(5％DMSO).于结扎后l、3、5和7 d(T1-4)时测定机械痛阈和热痛阈,T4时测完痛阈后处死大鼠取左侧L5节段脊髓背角,电镜下观察自噬体,采用Western blot法检测微管相关轻链蛋白3Ⅱ(LC3Ⅱ)和泛素结合蛋白p62表达,ELISA法检测IL-1β含量.结果与sham组比较,NP组大鼠各时点机械痛阈和T2-4时热痛阈降低,T4时脊髓背角LC3Ⅱ、p62表达水平及IL-1β含量升高(P＜0.05);与NP组比较,Rap组大鼠各时点机械痛阈和T2-4时热痛阈及T4时脊髓背角LC3Ⅱ表达水平升高,T4时脊髓背角p62表达水平及IL-1p含量降低(P＜0.05).电镜下NP组和Rap组大鼠脊髓背角可见自噬体,Rap组细胞器受损程度较NP组轻.结论大鼠神经病理性痛的形成与自噬障碍有关.
목적 평개자서재대서신경병이성통형성중적작용.방법 웅성SD대서30지,체중200 ～ 220 g,채용수궤수자표법분위3조(n=10):가수술조(sham조)、신경병이성통조(NP조)화자서유도제뢰파매소조(Rap조).행L4.5초내치관후,NP조화Rap조행좌측L5척신경결찰술제비대서신경병이성통모형,sham조대서부폭로좌측L5신경단불결찰.Rap조대서분별우결찰전30 min화술후2d초내주사뢰파매소60 μg,sham조화NP조주사등용용제(5％DMSO).우결찰후l、3、5화7 d(T1-4)시측정궤계통역화열통역,T4시측완통역후처사대서취좌측L5절단척수배각,전경하관찰자서체,채용Western blot법검측미관상관경련단백3Ⅱ(LC3Ⅱ)화범소결합단백p62표체,ELISA법검측IL-1β함량.결과 여sham조비교,NP조대서각시점궤계통역화T2-4시열통역강저,T4시척수배각LC3Ⅱ、p62표체수평급IL-1β함량승고(P＜0.05);여NP조비교,Rap조대서각시점궤계통역화T2-4시열통역급T4시척수배각LC3Ⅱ표체수평승고,T4시척수배각p62표체수평급IL-1p함량강저(P＜0.05).전경하NP조화Rap조대서척수배각가견자서체,Rap조세포기수손정도교NP조경.결론 대서신경병이성통적형성여자서장애유관.
Objective To evaluate the role of autophagy in the development of neuropathic pain (NP) in rats.Methods Thirty male Sprague-Dawley rats,weighing 200-220 g,were randomly divided into 3 groups (n =10 each) using a random number table:sham operation group (sham group),NP group and rapamycin (autophagy inducer) group (Rap group).Intrathecal catheter was inserted into the subarachnoid space and advanced caudally until the tip reached L4,5 segment.NP was induced by ligation of the left L5 spinal nerve (SNL) in NP and Rap groups.The L5 spinal nerve was only exposed,but not ligated in group sham.At 30 min before ligation and 2 days after operation,rapamycin 60 μg was injected intrathecally via the intrathecal catheter in Rap group,while the equal volume of vehicle (5％ dimethyl sulfoxide) was injected in sham and NP groups.The mechanical and thermal pain thresholds were measured on 1,3,5 and 7 days after ligation (T-4).The rats were sacrificed after the last measurement of pain threshold at T4.The ipsilateral L5 segment of spinal dorsal horn was removed for examination of autophagosomes (using transmission electron microscope) and for determination of the expression of LC3 Ⅱ and p62 (by Western blot) and content of IL-1β (by ELISA).Results Compared with sham group,the mechanical pain threshold at each time point and thermal pain threshold at T2-4 were significantly decreased,and the LC3 Ⅱ and p62 expression and IL-1β content were increased at T4 in group NP (P ＜ 0.05).Compared with NP group,the mechanical pain threshold at each time point,thermal pain threshold at T2-4 and LC3 Ⅱ expression at T4 were significantly increased,and the p62 expression and IL-1β content were decreased at T4 in group Rap (P ＜ 0.05).Microscopic examination showed that autophagosomes were observed in the spinal dorsal horn in NP and Rap groups,and the damage to organelles was lighter in Rap group than in NP group.Conclusion The development of NP is related to autophagic dysfunction in rats.