朱砂安神丸对大鼠VLPO脑区内单胺类递质的影响
|
摘要
目的:探讨朱砂安神丸的安神助眠作用机理,实时评价朱砂安神丸引起的单胺神经递质变化效应,及其对睡眠的影响。方法:采用微透析和高效液相色谱-电化学检测器(HPLC-ECD)联用方法测定了大鼠下丘脑腹外侧视前区(VLPO)中5-HT,DA,NE的24 h内的变化趋势。将探针插入大鼠右侧下丘脑腹外侧视前区(VLPO),大鼠在自由活动的清醒条件下,进行微透析操作,每隔2 h取样一次,24 h内连续取样。联合高效液相电化学HPLC-ECD进行样本分析,对VLPO脑区内的5-HT,DA,NE进行含量测定。结果:检测方法的线性范围为3. 125-50 pg﹒m L~(-1),日内和日间精密度均﹤3. 6,加标回收率为88. 8%-103. 8%,该方法适合于清醒动物脑内单胺类神经递质变化的测量分析。朱砂安神丸组NE,DA和5-HT在各时段均低于空白组。其中在7∶00-21∶00之间NE和5-HT含量显著低于空白组,P <0. 05有显著意义。DA在各时间段含量虽然给药组低于空白组,但变化不显著。除7∶00和17∶00两个时间段外,均无统计学意义。结论:朱砂安神丸抑制VLPO脑区5-HT和NE单胺类神经递质的含量,从而起到了安神助眠的作用。
Objective: To explore the mechanism of action of Zhusha Anshen pills and evaluate the effect of monoamine neurotransmitter changes caused by Zhusha Anshen Pills and its effects on sleep.Methods: The micro-dialysis and high performance liquid chromatography-electrochemical detector ( HPLC-ECD) method was used to determine the variation trend of 5-HT,DA and NE in the ventrolateral anterior optic region( VLPO) of rat hypothalamus within 24 hours. The probe was inserted into the right VLPO of the rat brain,and the rats were subjected to a microdialysis operation under freely awake conditions,sampled once every 2 hours,and continuously sampled within 24 hours. High-performance liquid phase electrochemistry was used to determine the content of 5-HT,DA,NE in the VLPO brain region. The results show that the linear range of the method is 3. 125-50 pg · mL~(-1),intraday RSD% and daytime RSD% are good,and the spiked recovery rate was 88. 8%-103. 8%. The method is suitable for measuring and analyzing the changes of monoamine neurotransmitters in the brain of awake animals.Results: The contents of NE,DA and 5-HT were lower in the drug-administered group than in the blank group at each time interval. Among them,NE and 5-HT levels were significantly lower than those in the blank group at 7-21 o'clock( P < 0. 05). Although the content of DA in each time period was lower than that in the blank group,the change was not significant. There was no statisticalsignificance except for the 7 o 'clock and 17 o 'clock periods.Conclusion: Zhusha Anshen Pill inhibits VLPO from accepting 5-HT and NE monoamine neurotransmitters,thus playing a role insoothingand helping the spleen.
Objective: To explore the mechanism of action of Zhusha Anshen pills and evaluate the effect of monoamine neurotransmitter changes caused by Zhusha Anshen Pills and its effects on sleep.Methods: The micro-dialysis and high performance liquid chromatography-electrochemical detector ( HPLC-ECD) method was used to determine the variation trend of 5-HT,DA and NE in the ventrolateral anterior optic region( VLPO) of rat hypothalamus within 24 hours. The probe was inserted into the right VLPO of the rat brain,and the rats were subjected to a microdialysis operation under freely awake conditions,sampled once every 2 hours,and continuously sampled within 24 hours. High-performance liquid phase electrochemistry was used to determine the content of 5-HT,DA,NE in the VLPO brain region. The results show that the linear range of the method is 3. 125-50 pg · mL~(-1),intraday RSD% and daytime RSD% are good,and the spiked recovery rate was 88. 8%-103. 8%. The method is suitable for measuring and analyzing the changes of monoamine neurotransmitters in the brain of awake animals.Results: The contents of NE,DA and 5-HT were lower in the drug-administered group than in the blank group at each time interval. Among them,NE and 5-HT levels were significantly lower than those in the blank group at 7-21 o'clock( P < 0. 05). Although the content of DA in each time period was lower than that in the blank group,the change was not significant. There was no statisticalsignificance except for the 7 o 'clock and 17 o 'clock periods.Conclusion: Zhusha Anshen Pill inhibits VLPO from accepting 5-HT and NE monoamine neurotransmitters,thus playing a role insoothingand helping the spleen.
引文
[1]唐霜.朱砂安神丸对条件性恐惧大鼠恐惧记忆的影响及作用机制研究[D].黑龙江中医药大学,2018.
[2]孙兵,郝洪谦.朱砂安神丸药理作用的实验研究[J].中成药,1995,(7):30-31.
[3]袁杰,喻田.全身麻醉机制与下丘脑腹外侧视前核睡眠通路[J].国际麻醉学与复苏杂志,2015,36(3):283-286.
[4]Sherin J E,Elmquist J K,Torrealba F,et al.Innervation of histaminergic tuberomammillary neurons by GABAergic and galaninergic neurons in the ventrolateral preoptic nucleus of the rat[J].J Neurosci,1998,18(12):4705-4721.
[5]Chou T C,Bjorkum A A,Gaus S E,et al.Afferents to the ventrolateral preoptic nucleus[J].J Neurosci,2002,22(3):977-990.
[6]丁丁,丁睿,吴芳,等.大鼠下丘脑腹外侧视前区和结节乳头体核的直接神经纤维投射研究[J].安徽医科大学学报,2014(10):1376-1378.
[7]岳小芳.腹外侧视前区腺苷及其受体对睡眠觉醒的调节作用及机制研究[D].复旦大学,2010.
[8]魏丽丽,陈虹,钟明,等.高效液相色谱-电化学法测定大鼠纹状体细胞外液中单胺类神经递质含量[J].药物分析杂志,2011(3):467-470.
[9]张如意.酸枣仁汤对基于5-HTR调控老年失眠SCN及VLPO系统的作用及机制研究[D].湖北中医药大学,2017.
[10]丁婉婷.基于伏隔核单胺类神经递质变化的酸枣仁改善睡眠作用机理的研究[D].黑龙江中医药大学,2017.
[11]何康.电刺激与电损毁腹外侧视前区对大鼠睡眠活动的影响[D].河北师范大学,2016.
[12]Saper C B,Scammell T E,Lu J.Hypothalamic regulation of sleep and circadian rhythms[J].Nature,2005,437(7063):1257.
[13]Saper C B,Fuller P M,Pedersen N P,et al.Sleep state switching.[J].Neuron,2010,68(6):1023-1042.
[14]吴芳.下丘脑腹外侧视前区与结节乳头体核对大鼠脑电活动的调节[D].安徽医科大学,2014.
[15]方光战.大鼠睡眠分期及脑电标度行为研究[D].电子科技大学,2010.
[16]王玳,申国华,李倩,等.朱砂安神丸的药用安全性评价与控制的思考[J].中国药品标准,2009,10(4):251-252.
[17]金阳,王广伟,李廷利.朱砂安神丸水煎剂对失眠大鼠睡眠时相的影响[J].上海中医药杂志,2008,42(12):74-76.
[18]袁杰,喻田.全身麻醉机制与下丘脑腹外侧视前核睡眠通路[J].国际麻醉学与复苏杂志,2015,36(3):283-286.
[2]孙兵,郝洪谦.朱砂安神丸药理作用的实验研究[J].中成药,1995,(7):30-31.
[3]袁杰,喻田.全身麻醉机制与下丘脑腹外侧视前核睡眠通路[J].国际麻醉学与复苏杂志,2015,36(3):283-286.
[4]Sherin J E,Elmquist J K,Torrealba F,et al.Innervation of histaminergic tuberomammillary neurons by GABAergic and galaninergic neurons in the ventrolateral preoptic nucleus of the rat[J].J Neurosci,1998,18(12):4705-4721.
[5]Chou T C,Bjorkum A A,Gaus S E,et al.Afferents to the ventrolateral preoptic nucleus[J].J Neurosci,2002,22(3):977-990.
[6]丁丁,丁睿,吴芳,等.大鼠下丘脑腹外侧视前区和结节乳头体核的直接神经纤维投射研究[J].安徽医科大学学报,2014(10):1376-1378.
[7]岳小芳.腹外侧视前区腺苷及其受体对睡眠觉醒的调节作用及机制研究[D].复旦大学,2010.
[8]魏丽丽,陈虹,钟明,等.高效液相色谱-电化学法测定大鼠纹状体细胞外液中单胺类神经递质含量[J].药物分析杂志,2011(3):467-470.
[9]张如意.酸枣仁汤对基于5-HTR调控老年失眠SCN及VLPO系统的作用及机制研究[D].湖北中医药大学,2017.
[10]丁婉婷.基于伏隔核单胺类神经递质变化的酸枣仁改善睡眠作用机理的研究[D].黑龙江中医药大学,2017.
[11]何康.电刺激与电损毁腹外侧视前区对大鼠睡眠活动的影响[D].河北师范大学,2016.
[12]Saper C B,Scammell T E,Lu J.Hypothalamic regulation of sleep and circadian rhythms[J].Nature,2005,437(7063):1257.
[13]Saper C B,Fuller P M,Pedersen N P,et al.Sleep state switching.[J].Neuron,2010,68(6):1023-1042.
[14]吴芳.下丘脑腹外侧视前区与结节乳头体核对大鼠脑电活动的调节[D].安徽医科大学,2014.
[15]方光战.大鼠睡眠分期及脑电标度行为研究[D].电子科技大学,2010.
[16]王玳,申国华,李倩,等.朱砂安神丸的药用安全性评价与控制的思考[J].中国药品标准,2009,10(4):251-252.
[17]金阳,王广伟,李廷利.朱砂安神丸水煎剂对失眠大鼠睡眠时相的影响[J].上海中医药杂志,2008,42(12):74-76.
[18]袁杰,喻田.全身麻醉机制与下丘脑腹外侧视前核睡眠通路[J].国际麻醉学与复苏杂志,2015,36(3):283-286.