刺五加注射液UPLC-MS成分分析及其抗抑郁作用研究  

Analysis of main chemical components of Ciwujia Injection based on UPLC-MS and study on its anti-depression effect

作　　者：严妍 李杏花 王旭 房城 吴修红 YAN Yan;LI Xinghua;WANG Xu;FANG Cheng;WU Xiuhong(Drug Safety Evaluation Center,Heilongjiang University of Chinese Medicine,Harbin 150040,China;College of Pharmacy,Heilongjiang University of Chinese Medicine,Harbin 150040,China)

机构地区：[1]黑龙江中医药大学药物安全性评价中心,黑龙江哈尔滨150040 [2]黑龙江中医药大学药学院,黑龙江哈尔滨150040

出　　处：《药物评价研究》2022年第7期1332-1342,共11页Drug Evaluation Research

基　　金：黑龙江省自然科学基金项目(LH2019H049);黑龙江省应用技术研究与开发计划(GA19C113)。

摘　　要：目的建立超高效液相色谱-串联质谱(UPLC-MS)分析方法,对刺五加注射液(CWJI)主要化学成分进行鉴定或表征;进一步研究CWJI对抑郁症模型大鼠行为学及神经肽水平的影响。方法采用优化后的UPLC-MS分析方法,在正负离子模式下提取质谱峰的一级质谱和二级质谱信息,鉴定CWJI中主要化学成分的化学结构。将雄性SD大鼠随机分为6组,分别为对照组、模型组、盐酸多塞平注射液(DH,阳性药,4.5 mg·kg^(−1))组和CWJI低、中、高剂量(44.1、88.2、132.3 mg·kg^(−1))组,除对照组外,其余各组均利用孤养结合慢性温和不可预知应激(28 d)的方法制备抑郁大鼠模型,在造模的同时开始ip给药,每天2次,每次间隔3 h。观察各组大鼠体质量、糖水偏好率、敞箱实验(水平穿越格数、直立次数、粪便粒数)的变化;采用放射免疫试剂盒法测定血浆中神经肽Y(NPY)、P物质(SP)、生长抑素(SS)水平。结果在CWJI中共鉴定出29个主要化学成分,包括刺五加苷D、刺五加苷E、异嗪皮啶、咖啡酸、绿原酸等。抗抑郁实验结果表明,与模型组比较,CWJI高剂量组与DH组在实验第21、28天大鼠体质量均显著升高(P<0.05、0.01);CWJI低、中、高剂量组及DH组大鼠的糖水偏好率显著增加(P<0.05、0.01);CWJI低、中、高剂量组及DH组大鼠的水平穿越格数均显著增加(P<0.01),中、高剂量组及DH组大鼠的直立次数均显著增多(P<0.01),粪便粒数均显著减少(P<0.05、0.01);CWJI低、中、高剂量组和DH组大鼠血浆中的NPY水平均显著升高,SP水平均显著降低(P<0.05、0.01),中、高剂量组和DH组的SS水平均显著升高(P<0.05、0.01)。结论CWJI具有显著的抗抑郁作用,其化学成分以苷类和黄酮类化合物为主,可能是其抗抑郁的主要活性成分。Objective Establish the analytical method of the main chemical components in Ciwujia Injection(CWJI),identify and characterize the main chemical components,and clarify the chemical basis from the whole.Further to investigate the antidepressant effects of CWJI on the behavior and neuropeptide level of depression rats induced by chronic unpredictable mild stress(CUMS).Methods The optimized UPLC-MS analysis methods were used to extract the first-order and second-order mass spectrometry information of peaks in positive and negative ion mode,and to identify the chemical structures of the main chemical components in CWJI.Male SD rats were randomly divided into six groups:control group,model group,doxepin hydrochloride injection(DH,positive drug,4.5 mg·kg^(−1))group and CWJI low-dose,medium-dose and high-dose(44.1,88.2,132.3 mg·kg^(−1))groups.The other groups were treated with solitary rearing combined with chronic unpredictable stress(28 d)to prepare the depressed rat model.Administration of ip was started at the same time of modeling,twice a day at an interval of 3 h.The changes of body weight,sugar water preference rate and open box experiment(number of horizontal crossing bars,number of vertical positions and fecal particles)were observed in each group.Plasma levels of neuropeptide Y(NPY),substance P(SP)and somatostatin(SS)were determined by radioimmunoassay.Results A total of 29 chemical constituents were identified in CWJI,including eleutheroside D,eleutheroside E,isofraxidin,caffeic acid,chlorogenic acid,etc.Compared with model group,the body weight of CWJI high-dose group and DH group was significantly increased at 21 and 28 days of experiment(P<0.05,0.01).The preference rate of sugar water in CWJI low,medium and high dose groups and DH group was significantly increased(P<0.05,0.01).The number of horizontal crossing bars in CWJI low-dose,medium-dose and high-dose groups and DH groups was significantly increased(P<0.01),the number of upright positions in CWJI low-dose,medium-dose and high-dose groups and DH

关 键 词：刺五加注射液 化学成分 刺五加苷D 刺五加苷E 异嗪皮啶 咖啡酸 绿原酸 慢性温和不可预知应激 抗抑郁 神经肽Y P物质 生长抑素 

分 类 号：R284.1[医药卫生—中药学] R285.5[医药卫生—中医学]