CAJ | 학술논문

目的探讨p38丝裂原活化蛋白激酶(p38MAPK)信号转导通路在脑组织fractalkine诱发小鼠痛觉过敏中的作用.方法雄性昆明小鼠225只,体重30～40 g,采用随机数字表法,将其随机分为4组:正常对照组(C组,n=55)、fractalkine组(F组,n=60)、CX3C趋化因子受体1(CX3CR1)抗体anti-CX3CR1+ fractalkine组(CF组,n=55)及p38MAPK抑制剂SB203580+ fractalkine组(SF组,n=55).F组、CF组及SF组经侧脑室注射fractalkine 100 ng,CF组及SF组给予fractalkine前1h时分别经侧脑室注射anti-CX3CR1 1μg和SB203580 1μg,C组给予等容量生理盐水.分别于侧脑室给药前30min、给予fractalkine后30、60、120、240 min时取10只小鼠,测定热缩爪潜伏期(PWL),然后于上述时点各处死小鼠5只,取脑组织,采用Western blot法测定p38 MAPK磷酸化水平.分别于侧脑室给药前30min、给予fractalkine后6、12、24h时处死小鼠5只,取脑组织,采用ELISA法测定IL-1β和TNF-α的含量.F组注射fractalkine后4h时处死5只小鼠,采用免疫荧光法确定fractalkinc作用于小胶质细胞或星形胶质细胞.结果与C组比较,其他3组PWL缩短,脑组织p38MAPK磷酸化、IL-1β和TNF-α水平升高(P＜0.05)；与F组比较,CF组及SF组PWL延长,脑组织p38MAPK磷酸化、IL-1β和TNF-α水平降低(P＜0.05).fractalkine作用于小胶质细胞.结论 p38MAPK信号转导通路参与了脑组织fractalkine诱发小鼠痛觉过敏的形成.
목적 탐토p38사렬원활화단백격매(p38MAPK)신호전도통로재뇌조직fractalkine유발소서통각과민중적작용.방법 웅성곤명소서225지,체중30～40 g,채용수궤수자표법,장기수궤분위4조:정상대조조(C조,n=55)、fractalkine조(F조,n=60)、CX3C추화인자수체1(CX3CR1)항체anti-CX3CR1+ fractalkine조(CF조,n=55)급p38MAPK억제제SB203580+ fractalkine조(SF조,n=55).F조、CF조급SF조경측뇌실주사fractalkine 100 ng,CF조급SF조급여fractalkine전1h시분별경측뇌실주사anti-CX3CR1 1μg화SB203580 1μg,C조급여등용량생리염수.분별우측뇌실급약전30min、급여fractalkine후30、60、120、240 min시취10지소서,측정열축조잠복기(PWL),연후우상술시점각처사소서5지,취뇌조직,채용Western blot법측정p38 MAPK린산화수평.분별우측뇌실급약전30min、급여fractalkine후6、12、24h시처사소서5지,취뇌조직,채용ELISA법측정IL-1β화TNF-α적함량.F조주사fractalkine후4h시처사5지소서,채용면역형광법학정fractalkinc작용우소효질세포혹성형효질세포.결과 여C조비교,기타3조PWL축단,뇌조직p38MAPK린산화、IL-1β화TNF-α수평승고(P＜0.05)；여F조비교,CF조급SF조PWL연장,뇌조직p38MAPK린산화、IL-1β화TNF-α수평강저(P＜0.05).fractalkine작용우소효질세포.결론 p38MAPK신호전도통로삼여료뇌조직fractalkine유발소서통각과민적형성.
Objective To determine whether p38 mitogen-activated protein kinase (p38MAPK) signaling pathway is involved in cerebral fractalkine-induced hyperalgesia in mice.Methods Two hundred and twenty-five male Kunming mice weighing 30-40 g were randomly divided into 4 groups:group control ( group C,n =55 ) ;group fractalkine (group F,n =60); group anti-CX3CR1 + fractalkine (group CF,n =55) and group SB203580 (p38MAPK inhibitor) + fractalkine (group SF,n =55).Fractalkine 100 ng was injected into cerebral lateral ventricle (i.c.v.) in groups F,CF and SF.Anti-CX3CR1 1 μg and SB203580 1 μg were injected i.c.v.at 1 h before fractalkine injection in groups CF and SF respectively.Paw withdrawal latency to a thermal nociceptive stimulus (PWL) was measured at 30 min before the drugs were injected into cerebral lateral ventricle and 30,60,120 and 240 min after fractalkine injection.Five animals were sacrificed after PWL measurement at each time point and their brains were removed for determination of phosphorylated p38MAPK protein expression (by Western blot analysis).Five animals were sacrificed at 30 min before the drugs were injected into cerebral lateral ventricle and 6,12 and 24 h after fractalkine injection for determination of IL-1β and TNF-α contents in the brain (by ELISA) in all the 4 groups.In group F 5 animals were sacrificed at 4 h after fractalkine injection for determination of action of fractalkine on microglia or astrocyte (by immunofluorescence).Results Fractalkine i.c.v.injection significantly reduced PWL and increased phosphorylated 38MAPK,IL-1β and TNF-α levels in group F as compared with group C.Pretreatment with anti-CX3CR1 or SB203580 significantly decreased fractalkine-induced hyperalgesia and phosphorylated-p38MAPK,IL-1β and TNF-α levels in groups CF and SF as compared with group F.Fractalkine was localized at microglia.Conclusion p38MAPK signal transduction pathway is involved in cerebral fractalkine-induced hyperalgesia in mice.