CAJ | 학술논문

目的评价鞘内注射雷帕霉素对大鼠糖尿病神经痛的影响.方法取鞘内置管成功的成年雄性SD大鼠30只,体重180～200 g,采用随机数字表法,将其分成5组(n=6):对照组(C组)、糖尿病神经痛组(DN组)、雷帕霉素1 μg组(R1组)、雷帕霉素3 μg组(R3组)和雷帕霉素10 μg组(R10组).鞘内置管第5天时DN组、R1组、R3组和R10组腹腔注射链脲霉素60 mg/kg制备糖尿病模型,C组腹腔注射枸橼酸-枸橼酸钠缓冲液6 ml/kg.于造模后21 d时,R1组、R3组和R10组分别鞘内注射雷帕霉素1、3、10 μg/d[溶于10 μl 4％二甲基亚砜(DMSO)],连续7d,C组和DN组鞘内注射4％ DMSO 10μl.于鞘内置管前、造模前、造模后7、14、21 d和鞘内给药1、3、5、7d时测定机械痛阈.痛阈测定结束后,处死大鼠,取L2-5节段脊髓,采用Western blot法测定雷帕霉素靶蛋白(mTOR)、磷酸化mTOR(p-mTOR)、核糖体蛋白S6激酶(S6K)和磷酸化S6K(p-S6K)的表达水平,采用RT-PCR法测定mTORmRNA和S6K mRNA的表达水平.结果与C组比较,DN组、R1组、R3组和R10组造模后14和21 d时机械痛阈降低,DN组脊髓mTOR、p-mTOR、S6K、p-S6K和mTOR mRNA、S6K mRNA表达上调(P＜0.01)；与DN组比较,R1组鞘内给药后5和7d时、R3组鞘内给药后3、5和7d时、R10组鞘内给药后1、3、5和7d时机械痛阈升高,R1组、R3组和R10组脊髓mTOR、p-mTOR、S6K、p-S6K和mTOR mRNA、S6K mRNA表达下调(P＜0.01).结论鞘内注射雷帕霉素可减轻大鼠糖尿病神经痛.
목적 평개초내주사뢰파매소대대서당뇨병신경통적영향.방법 취초내치관성공적성년웅성SD대서30지,체중180～200 g,채용수궤수자표법,장기분성5조(n=6):대조조(C조)、당뇨병신경통조(DN조)、뢰파매소1 μg조(R1조)、뢰파매소3 μg조(R3조)화뢰파매소10 μg조(R10조).초내치관제5천시DN조、R1조、R3조화R10조복강주사련뇨매소60 mg/kg제비당뇨병모형,C조복강주사구연산-구연산납완충액6 ml/kg.우조모후21 d시,R1조、R3조화R10조분별초내주사뢰파매소1、3、10 μg/d[용우10 μl 4％이갑기아풍(DMSO)],련속7d,C조화DN조초내주사4％ DMSO 10μl.우초내치관전、조모전、조모후7、14、21 d화초내급약1、3、5、7d시측정궤계통역.통역측정결속후,처사대서,취L2-5절단척수,채용Western blot법측정뢰파매소파단백(mTOR)、린산화mTOR(p-mTOR)、핵당체단백S6격매(S6K)화린산화S6K(p-S6K)적표체수평,채용RT-PCR법측정mTORmRNA화S6K mRNA적표체수평.결과 여C조비교,DN조、R1조、R3조화R10조조모후14화21 d시궤계통역강저,DN조척수mTOR、p-mTOR、S6K、p-S6K화mTOR mRNA、S6K mRNA표체상조(P＜0.01)；여DN조비교,R1조초내급약후5화7d시、R3조초내급약후3、5화7d시、R10조초내급약후1、3、5화7d시궤계통역승고,R1조、R3조화R10조척수mTOR、p-mTOR、S6K、p-S6K화mTOR mRNA、S6K mRNA표체하조(P＜0.01).결론 초내주사뢰파매소가감경대서당뇨병신경통.
Objective To evaluate the effect of intrathecal rapamycin on diabetic neuropathic pain in rats.Methods Thirty adult male Sprague-Dawley rats,weighing 180-220 g,in which IT catheters were successfully implanted,were randomly divided into 5 groups (n =6 each) using a random number table:control group (group C),diabetic neuropathic pain group (group DN),rapamycin 1 μg group (group R1),rapamycin 3 μg group (group R3) and rapamycin 10 μg (group R10).Diabetes mellitus was induced by intraperitoneal streptozotocin (STZ) 60 mg/kg on 5 days after IT catheters were implanted in DN,R1,R3 and R10 groups.Citric acid-sodium citrate buffer 6 ml/kg was injected intraperitoneally in group C.In R1,R3 and R10 groups,rapamycin (dissolved in 10 μl 4％ dimethyl sulfoxide) 1,3 and 10 μg were intrathecally injected,respectively,once a day for 7 consecutive days starting from day 21 after STZ injection,while the equal volume of 4％ dimethyl sulfoxide was given instead in C and DN groups.Paw withdrawal threshold (PWT) to yon Frey filament stimulation was measured before IT catheters were implanted,before STZ injection,on 7,14 and 21 days after STZ injection,and on 1,3,5 and 7 days after rapamycin administration.After measurement of PWT,the rats were sacrificed and L2-5 segments of the spinal cord were removed for determination of the expression of mTOR and phosphorylated mTOR (p-mTOR),S6K and phosphorylated S6K (p-S6K) (by Western blot) and expression of mTOR mRNA and S6K mRNA (by RT-PCR).Results Compared with group C,MWT was significantly decreased at 14 and 21 days after STZ injection in DN,R1,R3 and R10 groups,and the expression of mTOR,p-mTOR,S6K,p-S6K,mTOR mRNA and S6K mRNA was up-regulated in group DN (P ＜ 0.01).Compared with group DN,MWT was significantly increased at 5 and 7 days after rapamycin administration in group R1,at 3,5 and 7 days after rapamycin administration in group R3,and at 1,3,5 and 7 days after rapamycin administration in group R10,and the expression of mTOR,p-mTOR,S6K,p-S6K,mTOR mRNA and S6K mRNA was down-regulated in R1,R3 and R10 groups (P ＜ 0.01).Conclusion Intrathecal rapamycin can alleviate diabetic neuropathic pain in rats.