CAJ | 학술논문

目的观察盐酸戊乙奎醚(PHC)对吗啡依赖大鼠相关脑区p-CREB及M5受体的影响.方法将雄性SD大鼠30只随机分为对照组(NS组)、吗啡依赖组(MOR组)和PHC治疗组(PHC组),每组10只.连续7d交替皮下注射吗啡(10 mg/kg,每天1次)或生理盐水,诱导大鼠的吗啡依赖位置偏爱效应.实验第8天停用吗啡,NS组与MOR组腹腔注射等体积的生理盐水；PHC治疗组则腹腔注射PHC 1.5 mg/kg.30 min后各组大鼠行CPP测试.Western blot法检测大鼠中脑腹侧被盖区、伏隔核、前额叶皮层p-CREB的蛋白表达；实时定量聚合酶链反应(PCR)检测大鼠中脑腹侧被盖区、伏隔核、前额叶皮层M5受体的mRNA水平.结果 (1)PHC治疗组的灰区停留时间与MOR组比较显著缩短[(422±103)s 比(622 ±97)s,P＜0.01].(2)MOR组中脑腹侧被盖区、伏隔核、前额叶皮层组织中p-CREB水平显著高于NS组[(0.93±0.06)、(0.67±0.10)、(0.89±0.14)比(0.31±0.02)、(0.20±0.01)、(0.40±0.07),P＜0.01]；与MOR组比较,PHC组中脑腹侧被盖区、伏隔核、前额叶皮层组织中p-CREB水平显著降低[(0.65±0.05)、(0.58±0.04)、(0.67±0.09),P＜0.05或P＜0.01].(3)MOR组中脑腹侧被盖区、伏隔核、前额叶皮层组织中M5受体的mRNA含量较NS组显著增高(1.80、0.22、0.50比1.00、0.13、0.32,P＜0.01)；与MOR组比较,PHC 组能明显降低中脑腹侧被盖区、伏隔核、前额叶皮层组织中M5的mRNA含量(0.65、0.09、0.07,P＜0.01).结论 PHC能抑制吗啡依赖大鼠条件性位置偏爱的表达,该效应可能与PHC下调中脑腹侧被盖区、伏隔核、前额叶皮层组织中M5受体,抑制p-CREB表达有关.
목적 관찰염산무을규미(PHC)대마배의뢰대서상관뇌구p-CREB급M5수체적영향.방법 장웅성SD대서30지수궤분위대조조(NS조)、마배의뢰조(MOR조)화PHC치료조(PHC조),매조10지.련속7d교체피하주사마배(10 mg/kg,매천1차)혹생리염수,유도대서적마배의뢰위치편애효응.실험제8천정용마배,NS조여MOR조복강주사등체적적생리염수；PHC치료조칙복강주사PHC 1.5 mg/kg.30 min후각조대서행CPP측시.Western blot법검측대서중뇌복측피개구、복격핵、전액협피층p-CREB적단백표체；실시정량취합매련반응(PCR)검측대서중뇌복측피개구、복격핵、전액협피층M5수체적mRNA수평.결과 (1)PHC치료조적회구정류시간여MOR조비교현저축단[(422±103)s 비(622 ±97)s,P＜0.01].(2)MOR조중뇌복측피개구、복격핵、전액협피층조직중p-CREB수평현저고우NS조[(0.93±0.06)、(0.67±0.10)、(0.89±0.14)비(0.31±0.02)、(0.20±0.01)、(0.40±0.07),P＜0.01]；여MOR조비교,PHC조중뇌복측피개구、복격핵、전액협피층조직중p-CREB수평현저강저[(0.65±0.05)、(0.58±0.04)、(0.67±0.09),P＜0.05혹P＜0.01].(3)MOR조중뇌복측피개구、복격핵、전액협피층조직중M5수체적mRNA함량교NS조현저증고(1.80、0.22、0.50비1.00、0.13、0.32,P＜0.01)；여MOR조비교,PHC 조능명현강저중뇌복측피개구、복격핵、전액협피층조직중M5적mRNA함량(0.65、0.09、0.07,P＜0.01).결론 PHC능억제마배의뢰대서조건성위치편애적표체,해효응가능여PHC하조중뇌복측피개구、복격핵、전액협피층조직중M5수체,억제p-CREB표체유관.
Objective To explore the effect of penehyclidine hydrochloride (PHC) on the expression of phosphorylation of cyclic AMP response element binding protein (p-CREB) and M5 receptor of the relevant brain regions in morphine dependent rats.Methods Thirty male SD rats were randomly assigned to 3 groups ( n =10 each):morphine dependent group ( MOR group),PHC treatment groups ( PHC group),normal saline control group (NS group).The morphine conditioned place preference was induced by alternate subcutaneous injection of morphine for 7 days in rats (10 mg/kg,once daily,09:00) and saline (16:00).At the day 8,the rats received the CPP test.The rats in PHC group were treated with PHC (1.5 mg/kg,ip) prior to the CPP test,whereas the rats were treated with saline in MOR and NS group.Western blotting was used to detect the expression of p-CREB in the relevant brain regions ( including ventral tegmental area,nucleus accumbens,prefrontal cortex).Real-time polymerase chain reaction (PCR)was used to detect the expression of M5 receptor mRNA in the relevant brain regions ( including ventral tegmental area,nucleus accumbens,prefrontal cortex).Results ( 1 ) There were significant differences in the duration spent in the drug-paired side (gray area) between MOR and PHC groups [ (402 ± 103) vs.(622 ±97) s,P＜0.01] ; (2) In MOR group,the p-CREB levels in ventral tegmental area,nucleus accumbens and prefrontal cortex were significantly higher than those in NS group [ (0.93 ±0.06),(0.67 ±0.10),(0.89±0.14) vs.(0.31 ±0.02),(0.20±0.01),(0.40±0.07),P＜0.01].Acute treatment with PHC led to an obvious decrease of p-CREB level in ventral tegmental area,nucleus accumbens and prefrontal cortex [(0.65 ±0.05),(0.58 ±0.04),(0.67 ±0.09),P ＜0.05 or P＜0.01 ]; (3) As compared with NS group,the level of M5 receptor mRNA in ventral tegmental area,nucleus accumbens and prefrontal cortex was increased significantly during morphine dependence ( 1.80,0.22,0.50 vs.1.00,0.13,0.32,P ＜0.01 ).Treatment with PHC could significantly decrease the level of M5 receptor mRNA in ventral tegmental area,nucleus accumbens and prefrontal cortex (0.65,0.09,0.07,P＜0.01).Conclusion PHC could significantly inhibit the expression of CPP on morphine dependent rats,and this effect might be related to inhibition of the expression of p-CREB and down-regulation of M5 receptor in ventral tegmental area,nucleus accumbens and prefrontal cortex.