CAJ | 학술논문

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21 d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、DA 3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21 d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。
목적：건립측정대서혈장중오간색알(5-HT)、다파알(DA)、거갑신상선소(NE)적LC-MS/MS방법,관찰만성불가예견성온화응격( chronic unpredictable mild stress,CUMS)억욱대서혈장단알류신경체질DA、5-HT、NE적변화정황。방법♂ SD대서22지,분위대조조(n=10)화모형조(n=12),모형조매천급여9충만성불가예견성온화자격인자,조모21 d후,분별우조모전후진행행위학측정급안광채혈；채용분갑선록작위주전연생화시제,장3충대측물급내표연생화후진입LC-MS/MS검측,측정조모전후혈장중5-HT、NE、DA 3충신경체질농도。결과조모21d후,여대조조상비,모형조대서체중명현강저(P<0．05),수평득분、수직득분균명현강저(P <0．01),동시당수소모량명현강저(P <0．01)。혈장중5-HT、NE、DA적농도재1．47~752、1．75~898、2．05~1053μg·L-1범위내선성관계량호,최저정량한분별위1．47、1．75、2．05μg · L-1,이질공양품계산,재각농도수평하,차법적회수솔균대우70%,일내화일간정밀도소우15%,부합생물양품분석요구。조모21 d후,모형조혈장중5-HT、NE、DA적농도분별위(3．99±1．21)、(6．24±1．94)、(6．07±1．98)μg·L-1,여대조조상비균명현강저(P<0．01)。결론 CUMS조모성공후,혈장중적3충신경체질분별정하강추세。
Aim To establish a LC-MS/MS method for determination of 5-HT, NE, DA and observe the con-centration 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 given 9 kinds chronic unpredictable mild stimulating factors every day. 21 days later, behavior and orbital blood were measured before and after modeling. Using benzo-yl chloride as a pre-column derivatization reagent, three analytes and IS were derivatized before LC-MS/MS detection. Change in three kinds of neurotransmit-ter concentration was measured in rat plasma before and after modeling. Results After modeling, com-pared with control group, the weight of rats in model group was declined significantly ( P<0. 05 ) . Horizon-tal 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-1 and LOQ were 1. 47, 1. 75, 2. 046μg·L-1 ,respectively. The recov-ery of 5-HT, NE, DA from plasma was over than 70%, and RSD of inter-day and intra-day assay was limited in 15%. Compared with control group, the con-centration 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). Con-clusion After making CUMS model of depression, three kinds of neurotransmitters in rat plasma are de-creased.