CAJ | 학술논문

目的评价青藤碱对神经病理性痛大鼠脊髓背角神经元凋亡的影响.方法健康成年雄性Wistrar大鼠108只,体重180 ～ 220 g,采用随机数字表法分为3组(n=36):假手术组(S组)、神经病理性痛组(NP组)和青藤碱组(SIN组).NP组和SIN组采用坐骨神经慢性压迫性损伤法制备神经病理性痛模型,S组仅分离坐骨神经但不结扎.SIN组于术毕即刻开始至处死前1d,腹腔注射盐酸青藤碱注射液40 mg/kg,1次/d,S组和NP组腹腔注射等容量生理盐水.每组随机取12只大鼠于术前1d、术后3、7和14 d(T0-3)时测定机械痛阈和热痛阈,于T1-3时测定痛阈后处死大鼠,取L4-6脊髓组织,采用实时荧光定量PCR法检测脊髓背角caspase-3 mRNA和p38MAPK mRNA的表达,采用TUNEL法检测脊髓背角凋亡细胞,计算细胞凋亡率.结果与S组比较,NP组和SIN组热痛阈和机械痛阈降低,脊髓背角caspase-3 mRNA和p38MAPK mRNA表达上调,细胞凋亡率升高(P＜0.05);与NP组比较,SIN组热痛阈和机械痛阈升高,脊髓背角caspase-3 mRNA和p38MAPK mRNA表达下调,细胞凋亡率降低(P＜0.05);与T1时比较,NP组和SIN组T2.3时热痛阈和机械痛阈降低,脊髓背角caspase-3 mRNA和p38MAPK mRNA表达上调,细胞凋亡率升高(P＜0.05).结论青藤碱减轻大鼠神经病理性痛的机制与抑制p38MAPK信号传导通路,从而减少脊髓背角神经元凋亡有关.
목적 평개청등감대신경병이성통대서척수배각신경원조망적영향.방법 건강성년웅성Wistrar대서108지,체중180 ～ 220 g,채용수궤수자표법분위3조(n=36):가수술조(S조)、신경병이성통조(NP조)화청등감조(SIN조).NP조화SIN조채용좌골신경만성압박성손상법제비신경병이성통모형,S조부분리좌골신경단불결찰.SIN조우술필즉각개시지처사전1d,복강주사염산청등감주사액40 mg/kg,1차/d,S조화NP조복강주사등용량생리염수.매조수궤취12지대서우술전1d、술후3、7화14 d(T0-3)시측정궤계통역화열통역,우T1-3시측정통역후처사대서,취L4-6척수조직,채용실시형광정량PCR법검측척수배각caspase-3 mRNA화p38MAPK mRNA적표체,채용TUNEL법검측척수배각조망세포,계산세포조망솔.결과 여S조비교,NP조화SIN조열통역화궤계통역강저,척수배각caspase-3 mRNA화p38MAPK mRNA표체상조,세포조망솔승고(P＜0.05);여NP조비교,SIN조열통역화궤계통역승고,척수배각caspase-3 mRNA화p38MAPK mRNA표체하조,세포조망솔강저(P＜0.05);여T1시비교,NP조화SIN조T2.3시열통역화궤계통역강저,척수배각caspase-3 mRNA화p38MAPK mRNA표체상조,세포조망솔승고(P＜0.05).결론 청등감감경대서신경병이성통적궤제여억제p38MAPK신호전도통로,종이감소척수배각신경원조망유관.
Objective To evaluate the effects of sinomenine on neuronal apoptosis in the spinal dorsal horns in a rat model of neuropathic pain (NP).Methods One hundred and eight male Wistar rats,weighing 180-220 g,were randomly divided into 3 groups with 36 animals in each group:sham operation group (group S),NP group and sinomenine group (group SIN).The animals were anesthetized with intraperitoneal chloral hydrate.In groups NP and SIN,NP was induced by chronic constrictive injury.The sciatic nerve was exposed and four ligatures were placed on the right sciatic nerve at 1 mm intervals with 4-0 chromic catgut.In group S,the right sciatic nerves were exposed,but not ligated.In group SIN,sinomenine 40 mg/kg was injected intraperitoneally once a day starting from the end of operation until one day before the rats were sacrificed.In groups NP and S,the rats received the equal volume of normal saline instead of sinomenine.Twelve animals in each group were randomly chosen at 1 day before operation (T0) and 3,7 and 14 days after operation (T1-33) to measure mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL).The rats in each group were sacrificed at T1-3 after measurement of pain threshold and the lumbar segments (L4-6) of the spinal cord were removed for determination of caspase-3 mRNA and p38MAPK mRNA expression (by real-time fluorescent quantitative PCR) and cell apoptosis (by TUNEL) in the spinal dorsal horn.Apoptosis rate (AR =the number of apoptotic cells/the total number of cells examined) was calculated.Results Compared with group S,MWT and TWL were significantly decreased,the expression of caspase-3 mRNA and p38MAPK mRNA was up-regulated,and AR was increased at T1-3 in NP and SIN groups (P ＜ 0.05).Compared with group NP,MWT and TWL were significantly increased,the expression of caspase-3 mRNA and p38MAPK mRNA was down-regulated,and AR was decreased at T1-3 in group SIN (P ＜ 0.05).MWT and TWL were significantly lower,and the expression of caspase-3 mRNA and p38MAPK mRNA and AR were higher at T2,3 than at T1 in groups NP and SIN (P ＜ 0.05).Conclusion Sinomenine can attenuate NP through inhibiting the expression of p38MAPK signal transduction pathway and reducing neuronal apoptosis in the spinal dorsal horn of rats.