摘要
目的建立测定大鼠血浆中五羟色胺（5-HT）、多巴胺（DA）、去甲肾上腺素（NE）的LC-MS/MS方法,观察慢性不可预见性温和应激（chronic unpredictable mild stress,CUMS）抑郁大鼠血浆单胺类神经递质DA、5-HT、NE的变化情况.方法♂SD大鼠22只,分为对照组（n=10）和模型组（n=12）,模型组每天给予9种慢性不可预见性温和刺激因子,造模21d后,分别于造模前后进行行为学测定及眼眶采血;采用苯甲酰氯作为柱前衍生化试剂,将3种待测物及内标衍生化后进入LC-MS/MS检测,测定造模前后血浆中5-HT、NE、DA3种神经递质浓度.结果造模21d后,与对照组相比,模型组大鼠体重明显降低（P〈0.05）,水平得分、垂直得分均明显降低（P〈0.01）,同时糖水消耗量明显降低（P〈0.01）.血浆中5-HT、NE、DA的浓度在1.47～752、1.75～898、2.05～1053μg·L^-1范围内线性关系良好,最低定量限分别为1.47、1.75、2.05μg·L^-1,以质控样品计算,在各浓度水平下,此法的回收率均大于70%,日内和日间精密度小于15%,符合生物样品分析要求.造模21d后,模型组血浆中5-HT、NE、DA的浓度分别为（3.99±1.21）、（6.24±1.94）、（6.07±1.98）μg·L^-1,与对照组相比均明显降低（P〈0.01）.结论CUMS造模成功后,血浆中的3种神经递质分别呈下降趋势。
Aim To establish a LC-MS / MS method for determination of 5-HT,NE,DA and observe the concentration of 5-HT,NE,DA in rat plasma of CUMS.Methods Twenty-two male SD rats were divided into control group and model group. Model group was given9 kinds chronic unpredictable mild stimulating factorsevery day. 21 days later,behavior and orbital blood were measured before and after modeling. Using benzoyl chloride as a pre-column derivatization reagent,three analytes and IS were derivatized before LC-MS /MS detection. Change in three kinds of neurotransmitter concentration was measured in rat plasma before and after modeling. Results After modeling,compared with control group,the weight of rats in model group was declined significantly( P < 0. 05). Horizontal scores,vertical scores and sugar consumption were declined significantly( P < 0. 01). Calibration curves of 5-HT,NE,DA were linear between 1. 47 ~ 752,1. 75 ~ 898,2. 05 ~ 1 053 μg · L- 1and LOQ were1. 47,1. 75,2. 046 μg·L- 1,respectively. The recovery of 5-HT, NE, DA from plasma was over than70%,and RSD of inter-day and intra-day assay was limited in 15%. Compared with control group,the concentration of 5-HT,NE,DA in rat plasma of model group was declined to( 3. 99 ± 1. 21),( 6. 24 ±1. 94),( 6. 07 ± 1. 98) μg · L- 1( P < 0. 01). Conclusion After making CUMS model of depression,three kinds of neurotransmitters in rat plasma are decreased.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2015年第2期273-278,共6页
Chinese Pharmacological Bulletin
基金
国家科技部"重大新药创制"科技重大专项(No2012ZX09303009-002)
江苏省中医药领军人才(NoLJ200906)
江苏高校优学科建设工程资助项目(2010)
