CAJ | 학술논문

目的研究鞘内注射多巴胺D2受体激动剂喹吡罗对坐骨神经慢性压迫损伤大鼠脊髓水平胶质细胞源性神经营养因子表达的影响,探讨其介导抗伤害作用的可能机制. 方法实验1:采用完全随机分组方法将30只成年雄性SD大鼠制作坐骨神经慢性压迫损伤(chronic constriction injury of sciatic nerve,CCI)模型分为5组(每组6只):CCI+生理盐水(NS组)、CCI+喹吡罗0.1 μg(Q0.1组)、CCI+喹吡罗1μg(Q1组)、CCI+喹吡罗5 μg(Q5组)、CCI+喹吡罗10 μg(Q10组),分别在建模后第7天鞘内注射0.1、1、5、10 μg喹吡罗,NS组单次鞘内注射生理盐水10μl.于给药前及给药后0.5、1、2、4、8、16h测定大鼠术侧后足机械缩足反射阈值(paw withdrawal mechanical threshold,PWMT)和热缩足反射潜伏期(paw withdrawal thermal latency,PWTL).实验2:将54只成年雄性SD大鼠制作CCI模型,并采用完全随机分组方法分为3组(Q5组、Q10组、NS组,每组18只):Q5组、Q10组分别在建模后第7天鞘内注射5、10μg喹吡罗后0.5、1、2、4、8、16h处死取材,NS组单次鞘内注射生理盐水10μl;另取6只正常大鼠作为模型对照组(M组),另取6只正常大鼠作为对照组(C组).采用免疫印迹法测定脊髓背角胶质细胞源性神经营养因子(glial cell line-derived neurotrophic fact,GDNF)蛋白表达的变化. 结果实验1:与NS组比较,Q0.1组注药后各时间点PWMT和PWTL差异无统计学意义(P＞0.05),Q1组注药后2 h PWMT[(4.3±1.5)g]及PWTL[(13.2±1.6)s],Q5组注药后1,2、4 h PWMT[(4.7±1.6)、(5.3±1.6)、(4.7±2.1)g]和PWTL[(14.0±1.7)、(15.2±1.5)、(13.4±1.6)s],Q10组注药后1、2、4 h PWMT[(6.0±1.3)、(7.3±1.0)、(5.3±2.1)g]和PWTL [(15.3±1.8)、(17.5±1.2)、(14.9±1.7)s]明显升高(P＜0.05).实验2:与C组比较,M组GDNF表达(0.95±0.09)明显升高(P＜0.05).与M组和NS组比较,GDNF的表达Q5组[(1.47±0.12)、(1.24±0.05)]、Q10组[(1.63±0.08)、(1.27±0.06)]明显增加(P＜0.05),并且可以至少持续至注药后4h. 结论鞘内注射喹吡罗能够显著增加脊髓水平GDNF表达,同时伴有大鼠痛行为改善,GNDF的表达上调可能参与了多巴胺D2受体激动剂对神经病理性疼痛的调制过程. 目的研究鞘內註射多巴胺D2受體激動劑喹吡囉對坐骨神經慢性壓迫損傷大鼠脊髓水平膠質細胞源性神經營養因子錶達的影響,探討其介導抗傷害作用的可能機製. 方法實驗1:採用完全隨機分組方法將30隻成年雄性SD大鼠製作坐骨神經慢性壓迫損傷(chronic constriction injury of sciatic nerve,CCI)模型分為5組(每組6隻):CCI+生理鹽水(NS組)、CCI+喹吡囉0.1 μg(Q0.1組)、CCI+喹吡囉1μg(Q1組)、CCI+喹吡囉5 μg(Q5組)、CCI+喹吡囉10 μg(Q10組),分彆在建模後第7天鞘內註射0.1、1、5、10 μg喹吡囉,NS組單次鞘內註射生理鹽水10μl.于給藥前及給藥後0.5、1、2、4、8、16h測定大鼠術側後足機械縮足反射閾值(paw withdrawal mechanical threshold,PWMT)和熱縮足反射潛伏期(paw withdrawal thermal latency,PWTL).實驗2:將54隻成年雄性SD大鼠製作CCI模型,併採用完全隨機分組方法分為3組(Q5組、Q10組、NS組,每組18隻):Q5組、Q10組分彆在建模後第7天鞘內註射5、10μg喹吡囉後0.5、1、2、4、8、16h處死取材,NS組單次鞘內註射生理鹽水10μl;另取6隻正常大鼠作為模型對照組(M組),另取6隻正常大鼠作為對照組(C組).採用免疫印跡法測定脊髓揹角膠質細胞源性神經營養因子(glial cell line-derived neurotrophic fact,GDNF)蛋白錶達的變化. 結果實驗1:與NS組比較,Q0.1組註藥後各時間點PWMT和PWTL差異無統計學意義(P＞0.05),Q1組註藥後2 h PWMT[(4.3±1.5)g]及PWTL[(13.2±1.6)s],Q5組註藥後1,2、4 h PWMT[(4.7±1.6)、(5.3±1.6)、(4.7±2.1)g]和PWTL[(14.0±1.7)、(15.2±1.5)、(13.4±1.6)s],Q10組註藥後1、2、4 h PWMT[(6.0±1.3)、(7.3±1.0)、(5.3±2.1)g]和PWTL [(15.3±1.8)、(17.5±1.2)、(14.9±1.7)s]明顯升高(P＜0.05).實驗2:與C組比較,M組GDNF錶達(0.95±0.09)明顯升高(P＜0.05).與M組和NS組比較,GDNF的錶達Q5組[(1.47±0.12)、(1.24±0.05)]、Q10組[(1.63±0.08)、(1.27±0.06)]明顯增加(P＜0.05),併且可以至少持續至註藥後4h. 結論鞘內註射喹吡囉能夠顯著增加脊髓水平GDNF錶達,同時伴有大鼠痛行為改善,GNDF的錶達上調可能參與瞭多巴胺D2受體激動劑對神經病理性疼痛的調製過程.
목적 연구초내주사다파알D2수체격동제규필라대좌골신경만성압박손상대서척수수평효질세포원성신경영양인자표체적영향,탐토기개도항상해작용적가능궤제. 방법 실험1:채용완전수궤분조방법장30지성년웅성SD대서제작좌골신경만성압박손상(chronic constriction injury of sciatic nerve,CCI)모형분위5조(매조6지):CCI+생리염수(NS조)、CCI+규필라0.1 μg(Q0.1조)、CCI+규필라1μg(Q1조)、CCI+규필라5 μg(Q5조)、CCI+규필라10 μg(Q10조),분별재건모후제7천초내주사0.1、1、5、10 μg규필라,NS조단차초내주사생리염수10μl.우급약전급급약후0.5、1、2、4、8、16h측정대서술측후족궤계축족반사역치(paw withdrawal mechanical threshold,PWMT)화열축족반사잠복기(paw withdrawal thermal latency,PWTL).실험2:장54지성년웅성SD대서제작CCI모형,병채용완전수궤분조방법분위3조(Q5조、Q10조、NS조,매조18지):Q5조、Q10조분별재건모후제7천초내주사5、10μg규필라후0.5、1、2、4、8、16h처사취재,NS조단차초내주사생리염수10μl;령취6지정상대서작위모형대조조(M조),령취6지정상대서작위대조조(C조).채용면역인적법측정척수배각효질세포원성신경영양인자(glial cell line-derived neurotrophic fact,GDNF)단백표체적변화. 결과 실험1:여NS조비교,Q0.1조주약후각시간점PWMT화PWTL차이무통계학의의(P＞0.05),Q1조주약후2 h PWMT[(4.3±1.5)g]급PWTL[(13.2±1.6)s],Q5조주약후1,2、4 h PWMT[(4.7±1.6)、(5.3±1.6)、(4.7±2.1)g]화PWTL[(14.0±1.7)、(15.2±1.5)、(13.4±1.6)s],Q10조주약후1、2、4 h PWMT[(6.0±1.3)、(7.3±1.0)、(5.3±2.1)g]화PWTL [(15.3±1.8)、(17.5±1.2)、(14.9±1.7)s]명현승고(P＜0.05).실험2:여C조비교,M조GDNF표체(0.95±0.09)명현승고(P＜0.05).여M조화NS조비교,GDNF적표체Q5조[(1.47±0.12)、(1.24±0.05)]、Q10조[(1.63±0.08)、(1.27±0.06)]명현증가(P＜0.05),병차가이지소지속지주약후4h. 결론 초내주사규필라능구현저증가척수수평GDNF표체,동시반유대서통행위개선,GNDF적표체상조가능삼여료다파알D2수체격동제대신경병이성동통적조제과정.
Objective To study the effects of intrathecal injection of dopamine D2 receptor agonist quinpirole on expression of glial cell line-derived neurotrophic factor in the spinal cord in rats with chronic constrictive injury (CCI) and explore its possible mechanism of mediating antinociception.Methods In this study,models were established CCI in male Sprague-Dawley rats.The experiment was performed as 2 parts.In part one,30 CCI rats were randomly divided into 5 groups (n=6):CCI+saline (group NS),CCI+quinpirole 0.1 μg (group Q0.1),CCI+quinpirole 1 μg (group Q1),CCI+quinpirole 5 μg (group Q5),CCI+quinpirole 10μg (group Q 10).The drugs were injected intrathecally on day 7 after CCI,respectively.Paw withdrawal mechanical threshold (PWMT) and paw withdrawal thermal latency (PWTL) were measured before and at 0.5,1,2,4,8 h and 16 h after intrathecal injection.In part two 54 CCI rats were randomly divided into 3 groups (group Q5,Q10 and NS,n=18).Group Q5 and Q10 were sacrificed at 0.5,1,2,4,8,16 h after intrathecal quinpirole 5,10 μg on day 7 after CCI.Group NS,a single intrathecal injection of saline 10 μl.Group M,another 6 normal rats as model control group.Another 6 normal rats as control group C.The expression of glial cell linederived neurotrophic fact (GDNF) in the spinal cord was determined by Western blot.Results Part one:compared with group NS,the PWMT and PWTL of group Q0.1 at each time point after injection had no statistically significant difference (P＞0.05).The PWMT and PWTL of 2 h of group Q1 [(4.3±1.5) g,(13.2±1.6) s].1,2,4 h of group Q5 [(4.7±1.6),(5.3±1.6),(4.7±2.1) g,(14.0±1.7),(15.2±1.5),(13.4±1.6) s] and Q10 [(6.0±1.3),(7.3±1.0),(5.3±2.1) g,(15.3±1.8),(17.5±1.2),(14.9±1.7) s] after drug administration were significantly promoted (P＜0.05).Part two:compared with group C,the expression of GDNF was significantly increased in group M (0.95±0.09)(P＜0.05).Compared with group M and NS,group Q5 [(1.47±0.12),(1.24±0.05)] and Q10[(1.63±0.08),(1.27±0.06)] significantly increased the expression of GDNF (P＜0.05),and it can last until 4 h after injection at least.Conclusions Intrathecal injection of quinpirole can significantly increase the level of GDNF expression in the spinal cord.At the same time,the pain behavior was improved.The upregulation of GNDF expression is involved in the modulation of dopamine D2 receptor agonist on neuropathic pain.