CAJ | 학술논문

目的观察脊髓内源性硫化氢(H2S)在大鼠糖尿病神经痛维持中的作用.方法雄性SD大鼠(体质量150～170 g)48只,其中24只大鼠腹腔注射链脲霉素(60 mg/kg)制备成糖尿病神经痛(DNP)模型,其余24只大鼠腹腔注射等体积生理盐水作为对照.48只大鼠均于腹腔注射2周后行鞘内置管,然后将DNP模型大鼠随机分为两组(n=12):D组和DM组,将正常对照大鼠分成两组(n=12):C组和CM组;腹腔注射后3周,C组、D两组鞘内给予生理盐水10μl;CM、DM组鞘内给予10 μmol/L高铁血红蛋白10μl,每日1次,连续4d,分别于鞘内注射第1天(T1)、第2天(T2)、第3天(T3)、第4天(T4)4个时间点,在给药前后30 min测定各组大鼠50％机械缩足阈值(PWT),并于T4测量结束后处死大鼠,取腰骶膨大处脊髓,测定脊髓背角N-甲基-D-天冬氨酸受体2B亚基(NR2B) mRNA和蛋白表达.结果与C组和CM组比较,D组和DM组大鼠PWT降低,NR2B mRNA和蛋白表达上调(P＜0.05);与D组PWT比较,DM组大鼠在T2～T4时间点的给药前、T1 ～T4时间点的给药后PWT升高;与D组mRNA(0.89±0.09)和蛋白表达(1.28 ±0.17)比较,DM组大鼠脊髓背角NR2B mRNA(0.69 ±0.11)和蛋白表达(0.98 ±0.14)下调(P＜0.05);与给药前比较(3.8±0.9、6.1±1.4),DM组大鼠在T1～T2时间点给药后PWT(7.5±1.8、8.7±1.6)升高(P＜0.05).结论脊髓内源性H2S参与了大鼠糖尿病神经痛的维持,其机制与上调脊髓背角NR2B表达有关.
목적 관찰척수내원성류화경(H2S)재대서당뇨병신경통유지중적작용.방법 웅성SD대서(체질량150～170 g)48지,기중24지대서복강주사련뇨매소(60 mg/kg)제비성당뇨병신경통(DNP)모형,기여24지대서복강주사등체적생리염수작위대조.48지대서균우복강주사2주후행초내치관,연후장DNP모형대서수궤분위량조(n=12):D조화DM조,장정상대조대서분성량조(n=12):C조화CM조;복강주사후3주,C조、D량조초내급여생리염수10μl;CM、DM조초내급여10 μmol/L고철혈홍단백10μl,매일1차,련속4d,분별우초내주사제1천(T1)、제2천(T2)、제3천(T3)、제4천(T4)4개시간점,재급약전후30 min측정각조대서50％궤계축족역치(PWT),병우T4측량결속후처사대서,취요저팽대처척수,측정척수배각N-갑기-D-천동안산수체2B아기(NR2B) mRNA화단백표체.결과 여C조화CM조비교,D조화DM조대서PWT강저,NR2B mRNA화단백표체상조(P＜0.05);여D조PWT비교,DM조대서재T2～T4시간점적급약전、T1 ～T4시간점적급약후PWT승고;여D조mRNA(0.89±0.09)화단백표체(1.28 ±0.17)비교,DM조대서척수배각NR2B mRNA(0.69 ±0.11)화단백표체(0.98 ±0.14)하조(P＜0.05);여급약전비교(3.8±0.9、6.1±1.4),DM조대서재T1～T2시간점급약후PWT(7.5±1.8、8.7±1.6)승고(P＜0.05).결론 척수내원성H2S삼여료대서당뇨병신경통적유지,기궤제여상조척수배각NR2B표체유관.
Objective To evaluate the role of endogenous hydrogen sulfide in the maintenance of diabetic neuropathic pain (DNP) in rats.Methods The DNP models were established by intraperitoneal injection of stretopzin (STZ) in 24 male SD rats aged 4 weeks (150-170 g),and other 24 rats were intraperitoneally injected with the same volume of normal saline as normal group.All the rats were implanted intrathecal catheter.The rats with DNP were randomly divided into 2 subgroups (n =12 each):saline group (group D) and methemoglobin group (group DM).The rats in normal group were randomly divided into 2 subgroups (n =12 each):saline group (group C) and methemoglobin group (group CM).Three weeks after intraperitoneal injection,the rats were separately intrathecally administered methemoglobin 10 μ mol/L (10 μ1) (group DM),saline 10 μl (group D),methemoglobin 10 μmol/L (10 μl) (group CM) and saline 10 μl (group C),respectively,once a day for 4 days.The paw withdrawal mechanical threshold (PWT) of the rats were measured before and after injection at 1 (T1),2 (T2),3 (T3),4 (T4) days,then the rats were sacrificed at T4 point after the measurement and the lumbar segments of the spinal cord were removed for determination of N-methyl-D-aspartate receptor subunit 2B (NR2B) protein expression (by Western blotting) and NR2B mRNA expression [by reverse transcriptase-polymerase chain reaction (RT-PCR)].Results As compared with group C and group CM,the PWT in group D and group DM was significantly reduced (P ＜ 0.05),while the expression of NR2B protein and mRNA was significantly increased (P ＜ 0.05).As Compared with group D [NR2B mRNA (0.89 ± 0.09),NR2B protein (1.28 ± 0.17)],the PWT in group DM was significantly increased at T2-T4 point before the injection and T1-T4 point after the injection,while the expression of NR2B mRNA (0.69 ±0.11) and NR2B protein (0.98 ± 0.14) in group DM was significantly decreased (P ＜ 0.05).As compared with the point before the injection (3.8 ± 0.9 and 6.1 ± 1.4),the PWT in group DM were significantly increased at T1-T2 point after the injection (7.5 ± 1.8 and 8.7 ± 1.6) (P ＜ 0.05).Conclusion Endogenous hydrogen sulfide in the spinal cord is involved in the maintenance of DNP in rats and the possible mechanisms were related with up-regulation of NR2B in the spinal dorsal horn.