[關(guān)鍵詞]
[摘要]
目的 建立超高效液相色譜-串聯(lián)質(zhì)譜(UPLC-MS)分析方法,對刺五加注射液(CWJI)主要化學(xué)成分進(jìn)行鑒定或表征;進(jìn)一步研究CWJI對抑郁癥模型大鼠行為學(xué)及神經(jīng)肽水平的影響。方法 采用優(yōu)化后的UPLC-MS分析方法,在正負(fù)離子模式下提取質(zhì)譜峰的一級質(zhì)譜和二級質(zhì)譜信息,鑒定CWJI中主要化學(xué)成分的化學(xué)結(jié)構(gòu)。將雄性SD大鼠隨機分為6組,分別為對照組、模型組、鹽酸多塞平注射液(DH,陽性藥,4.5 mg·kg-1)組和CWJI低、中、高劑量(44.1、88.2、132.3 mg·kg-1)組,除對照組外,其余各組均利用孤養(yǎng)結(jié)合慢性溫和不可預(yù)知應(yīng)激(28 d)的方法制備抑郁大鼠模型,在造模的同時開始ip給藥,每天2次,每次間隔3 h。觀察各組大鼠體質(zhì)量、糖水偏好率、敞箱實驗(水平穿越格數(shù)、直立次數(shù)、糞便粒數(shù))的變化;采用放射免疫試劑盒法測定血漿中神經(jīng)肽Y (NPY)、P物質(zhì)(SP)、生長抑素(SS)水平。結(jié)果 在CWJI中共鑒定出29個主要化學(xué)成分,包括刺五加苷D、刺五加苷E、異嗪皮啶、咖啡酸、綠原酸等??挂钟魧嶒灲Y(jié)果表明,與模型組比較,CWJI高劑量組與DH組在實驗第21、28天大鼠體質(zhì)量均顯著升高(P<0.05、0.01); CWJI低、中、高劑量組及DH組大鼠的糖水偏好率顯著增加(P<0.05、0.01); CWJI低、中、高劑量組及DH組大鼠的水平穿越格數(shù)均顯著增加(P<0.01),中、高劑量組及DH組大鼠的直立次數(shù)均顯著增多(P<0.01),糞便粒數(shù)均顯著減少(P<0.05、0.01); CWJI低、中、高劑量組和DH組大鼠血漿中的NPY水平均顯著升高,SP水平均顯著降低(P<0.05、0.01),中、高劑量組和DH組的SS水平均顯著升高(P<0.05、0.01)。結(jié)論 CWJI具有顯著的抗抑郁作用,其化學(xué)成分以苷類和黃酮類化合物為主,可能是其抗抑郁的主要活性成分。
[Key word]
[Abstract]
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 groups was significantly increased (P<0.01), and the fecal grains were significantly decreased (P<0.05, 0.01). The plasma level of NPY and SP in CWJI low, medium and high dose groups and DH groups were significantly increased (P<0.05, 0.01), and the plasma level of SS in CWJI low, medium and high dose groups and DH groups was significantly increased (P<0.05, 0.01). Conclusion CWJI had antidepressant effects, and its chemical components were mainly glycosides and flavonoids, which were the main active ingredients of anti-stress reaction, anti-fatigue reaction and anti-oxidation reaction.
[中圖分類號]
R284.1;R285.5
[基金項目]
黑龍江省自然科學(xué)基金項目(LH2019H049);黑龍江省應(yīng)用技術(shù)研究與開發(fā)計劃(GA19C113)