朱砂安神丸对大鼠VLPO脑区内单胺类递质的影响

The effect of Zhusha Anshen Pill on monoamine neurotransmitters in rat VLPO brain region

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.