木果楝柠檬苦素的质谱分析与抗抑郁研究
发布时间：2018-08-25 16:02
【摘要】：在发现先导化合物用于开发具有治疗人类疾病的药物过程中,天然产物具有主导地位。事实上,植物和微生物的药理活性成分是新药研发的重要途径。木果楝(Xylocarpus granatum Koenig)具有多种活性,并且近年被广泛证明。但是对其抗抑郁活性的研究未见报道。快速、灵敏的质谱技术在天然产物中的应用越来越广泛,且质谱裂解规律有助于快速识别并解析化合物的结构,而对已知物去重复,进而探索未知化合物进行深入研究。目前,关于木果楝中含有独特结构的柠檬苦素的质谱研究还鲜有报道。因此,本研究运用质谱方法对木果楝柠檬苦素类成分进行鉴定和定量的快速识别,并对木果楝柠檬苦素的抗抑郁活性进行了初步实验研究,本研究为从木果楝中快速发现新的化合物及鉴定和质量控制木果楝奠定基础,为从木果楝柠檬苦素中发现具有抗抑郁活性先导化合物提供实验依据。第一部分木果楝柠檬苦素的质谱鉴定目的:通过分析木果楝柠檬苦素的二级质谱(Q-TOF-MS/MS),探索其质谱裂解途径,总结裂解规律。运用超高效液相色谱串联四级杆飞行时间质谱(UHPLC-Q-TOF-MS/MS)对木果楝柠檬苦素类化合物进行鉴定。方法:1 UHPLC-Q-TOF-MS/MS分析条件:Kinetex-C18色谱柱(100 mm×2.1mm,2.6μm),柱温:40℃。流动相:A为乙腈,B为水-0.1%甲酸-1 m M乙酸铵溶液。梯度洗脱,洗脱程序如下:10%A(0~5 min);10-30%A(5~10min);30-35%A(10~20 min);35-50%A(20~35 min);50-95%A(35~45min);95%A(45~50 min)。流速:0.4 m L/min。进样量:2μL。Duo Spray电喷雾离子(ESI)源;离子源温度(turbo spray temperature):600℃。气帘气(CUR):35 psi;雾化气(Gas 1):50 psi;加热气(Gas 2):50 psi;在正离子模式下,喷雾电压(ion spray voltage)为5500 V,解簇电压(DP)为80 V。信息相关采集模式,包括TOF-MS探测扫描100-1000 Da(200ms)和最多8个依赖的TOF-MS/MS扫描50-1000 Da(100 ms),碰撞能量为35 e V,碰撞扩展能量为±15 e V。2鉴定策略:经验分子式的计算和碎片离子的预测使用Peak View?软件完成。通过与对照品比较保留时间、精确质量和MS/MS质谱,明确鉴定已知化合物。基于质谱裂解规律、精确质量、保留时间和植物化学文献,初步鉴定已知化合物和推测鉴定未知化合物。结果:1应用Q-TOF-MS/MS分析并总结了17个木果楝柠檬苦素的质谱裂解规律。这些裂解规律主要包括,丢失酰基或酰氧基侧链、丢失水分子及CO分子。2应用UHPLC-Q-TOF-MS/MS鉴定了木果楝柠檬苦素类化合物。从木果楝果实提取物中检测到了至少300个化合物。通过与对照品对比,明确鉴定了17个已知化合物。初步鉴定了116个化合物。鉴定了7个潜在的新化合物。结论:1首次系统的研究了17个木果楝柠檬苦素的质谱裂解规律。2 UHPLC-Q-TOF-MS/MS法鉴定了木果楝柠檬苦素类化合物。基于对照品的质谱裂解规律、植物化学文献,推测鉴定了木果楝果实中的柠檬苦素类化合物。UHPLC-Q-TOF-MS/MS法鉴定木果楝柠檬苦素简单、快速、灵敏度高、专属性好。鉴定这些化合物为进一步研究木果楝中柠檬苦素类化合物的植物化学、代谢物鉴定和质量控制提供了重要依据。第二部分木果楝柠檬苦素的质谱定量测定目的:开发和验证一种化学计量学辅助的LC-MS/MS方法用于测定17种柠檬苦素,揭示木果楝果实不同部位柠檬苦素的化学组成与分布。方法:1 LC-MS/MS分析条件:17个分析物的LC分离在40℃的Phenomenex Kinetex柱(XB-C18,100 mm×2.1 mm,2.6μm)中进行,使用5μL进样体积。流动相由乙腈(A)和0.1%(v/v)甲酸(B)组成,流速为300μL/min。以下梯度程序用于定量:0.0-1.0 min,45-55%A;1.0-4.0 min,55-60%A;4.0-5.0 min,60-95%A;5.0-10.0 min,95%A;10.0-11.0 min,95-45%A。ESI喷雾电压为5500 V,正离子模式监测,喷雾温度为600℃。气帘气、雾化气体(Gas 1)和加热器气体(Gas 2)分别设置在30、50和55 psi。在所有情况下使用氮气。预设的MRM算法用于在90 s的MRM检测窗口和0.7 s的目标扫描时间,在预期的保留时间监测34个离子对(每个化合物2个离子对)。在增强产物离子(enhanced product ion,EPI)模式中,信息相关采集(information dependent acquisition,IDA)扫描触发相关的MS/MS质谱扫描以便更好地确认被测物。将完整的分子指纹信息保存在EPI谱中,显著降低假阳性和阴性结果的风险。使用35 e V的标准化碰撞能(collision energy,CE)以15 e V的扩展碰撞能量在4000 Da/s的扫描速度下产生EPI质谱。IDA设置中,选择响应信号最高的前体离子(质量差异范围为250 m Da),每个探测扫描都扣除动态背景,信号阈值为100cps。应用Master View?软件进行EPI鉴定,纯度值设置在大于等于70%为阳性结果。常规MRM与预设MRM相比,驻留时间是唯一的不同参数,它用20 ms替换了每对离子的监测时间窗口。2对预设MRM LC-MS/MS方法进行验证,包括,线性、最低检测限、精密度、重复性、稳定性、回收率和基质效应。响应面优化方法用于优化超声辅助提取。主成分分析(principal component analysis,PCA)和正交偏最小二乘法判别分析(orthogonal partial least squares discriminant analysis,OPLS-DA)用于分析LC-MS/MS数据,可提供化学计量学模型用于简便的可视化和解释木果楝果实不同部位的差异。结果:1本研究中,开发和验证了化学计量学辅助的LC-MS/MS法同时测定木果楝果实不同部位的17种柠檬苦素。与常规MRM相比,预设MRM提高了灵敏度和精度。预设MRM LC-MS/MS方法得到了良好的验证,包括,线性(R0.99)、最低检测限(0.20-10.0 ng/m L)、精密度(日内1.2%-7.6%和日间1.1%-2.6%)、重复性(2.1%-6.9%)、稳定性(4.3%-7.2%)、回收率(88.2%-106.2%)和基质效应(90.1%-103.3%)。2通过RSM获得的最优超声辅助提取条件比一般单次单因素法更准确。该方法用于分析35批木果楝果实不同部位的柠檬苦素。所有17种柠檬苦素显示明显的排序,种仁种子果实;13种柠檬苦素显示另一种排序,种仁种子果实果皮种皮。基于LC-MS/MS数据的PCA和OPLS-DA分析,确定了木果楝果实不同部分分类的潜在化学标记物。结论:本研究中,为了揭示木果楝果实不同部位柠檬苦素的化学成分组成与分布,开发和验证了化学计量学辅助的LC-MS/MS法同时测定17种柠檬苦素。另外,本研究表明针对LC-MS/MS进行多组分定量的开发、优化和数据分析,由RSM,PCA和OPLS-DA组成的化学计量学辅助的综合策略是有效和可行的。此外,本研究提供了鉴定和质量控制木果楝果实作为民间药物的化学成分组成与分布的证据。第三部分木果楝柠檬苦素的抗抑郁活性初步实验研究目的:研究木果楝柠檬苦素在急性应激诱导的小鼠抑郁模型中潜在的抗抑郁活性和作用机制。方法:通过行为学实验研究木果楝柠檬苦素xylocarpin H、cipadesin A和xyloccensin K的抗抑郁活性,通过神经内分泌系统研究其潜在的作用机制。木果楝柠檬苦素xylocarpin H、cipadesin A和xyloccensin K的抗抑郁活性(5,15,50 mg/kg/day for 7days,intragastrically)通过强迫游泳实验、悬尾实验和旷场实验评价。通过酶免疫法测定血清皮质酮和促肾上腺皮质激素,评价xylocarpin H和cipadesin A对下丘脑-垂体-肾上腺轴的作用。结果:木果楝柠檬苦素能显著降低小鼠强迫游泳实验中的漂浮不动时间(xylocarpin H、cipadesin A和xyloccensin K在15-50 mg/kg),显著降低小鼠悬尾实验中悬尾不动时间(xylocarpin H和cipadesin A在15-50mg/kg,xyloccensin K在5-50 mg/kg)。木果楝柠檬苦素显著增加小鼠旷场实验中在旷场中央区域停留时间(xylocarpin H在5-50 mg/kg,cipadesin A在15-50 mg/kg,xyloccensin K在50 mg/kg),并且不影响自发活动。另外,重复给予xylocarpin H和cipadesin A显著降低了处于强迫游泳实验小鼠血清中皮质酮CORT(xylocarpin H在15-50 mg/kg,cipadesin A在5-50 mg/kg)和促肾上腺皮质激素ACTH(xylocarpin H和cipadesin A在15-50 mg/kg)的水平,但对于未经应激处理的小鼠无影响。结论:木果楝柠檬苦素xylocarpin H、cipadesin A和xyloccensin K在急性应激诱导的小鼠抑郁模型中具有潜在的抗抑郁活性,潜在的作用机制可能是通过抑制下丘脑-垂体-肾上腺轴对应激应答的活性。这些数据支持对开发木果楝柠檬苦素作为潜在的抗抑郁和焦虑药物做进一步的研究。
[Abstract]:Natural products play a leading role in the discovery of lead compounds for the development of drugs for the treatment of human diseases. In fact, the pharmacological active ingredients of plants and microorganisms are important approaches for the development of new drugs. Rapid and sensitive mass spectrometric techniques have been widely used in natural products. Mass spectrometric pyrolysis is helpful to identify and analyze the structures of compounds quickly, and repeat the known compounds to explore the unknown compounds. Mass spectrometry is rarely reported. Therefore, this study used mass spectrometry to identify and quantify the limonin components of Melia azedarach, and carried out a preliminary experimental study on the antidepressant activity of Melia azedarach limonin. This study laid a foundation for the rapid discovery of new compounds from Melia azedarach and the identification and quality control of Melia azedarach. The first part is the mass spectrometric identification of limonin from Melia azedarach. Objective: By analyzing the secondary mass spectrometry of limonin from Melia azedarach (Q-TOF-MS/MS), to explore its mass spectrometric cleavage pathway and summarize the cleavage rule. Methods: 1 UHPLC-Q-TOF-MS/MS analysis conditions: Kinetex-C18 column (100 m m *2.1 m m, 2.6 micron), column temperature: 40 C. Mobile phase: acetonitrile A, water-0.1% formic acid-1 M ammonium acetate solution. Gradient elution procedure: 10% A (0-5 min); % A (5-10 min); 30-35% A (10-20 min); 35-50% A (20-35 min); 50-95% A (35-45 min); 95% A (45-50 min). Flow rate: 0.4 m L/min. Sample volume: 2 mu L. Duo Spray Electrospray Ion Source (ESI); Turbospray temperature: 600 (?) CUR: 35 psi; Gas 1: 50 psi; Heating gas (50 psi); Spray: 50 psi in positive ion mode The voltage (ion spray voltage) is 5500 V and the unwrapping voltage (DP) is 80 V. Information correlation acquisition mode includes TOF-MS probe scan 100-1000 Da (200 ms) and up to 8 dependent TOF-MS/MS scan 50-1000 Da (100 ms), collision energy 35 e V, collision spread energy (+15 e V.2) identification strategy: empirical molecular formula calculation and debris ion prediction enabled The known compounds were identified by comparing retention time, accurate mass and MS/MS mass spectrometry with the reference materials. Based on the mass spectrometric pyrolysis law, accurate mass, retention time and phytochemical literature, the known compounds were identified and the unknown compounds were speculated. Mass spectrometric fragmentation of Limonin in Melia azedarach fruit. These fragmentation patterns mainly include loss of acyl or acyloxy side chains, loss of water and CO molecules. 2 UHPLC-Q-TOF-MS/MS was used to identify limonin compounds in Melia azedarach fruit. At least 300 compounds were detected from the extracts of Melia azedarach fruit. The results were compared with those of the control. Seventeen known compounds were identified. Seven potential new compounds were identified. CONCLUSION: 1. Mass spectrometric fragmentation of 17 limonins from Melia azedarach was systematically studied for the first time. 2 U-HPLC-Q-TOF-MS/MS methods were used to identify the limonins from Melia azedarach. The identification of limonoids in the fruit of Melia azedarach by UHPLC-Q-TOF-MS/MS is simple, rapid, sensitive and specific. The identification of these compounds provides an important basis for further study on the phytochemistry, metabolites identification and quality control of the limonoids in Melia azedarach. Objective: To develop and validate a chemometric-assisted LC-MS/MS method for the determination of 17 kinds of Limonin in Melia azedarach fruits, and to reveal the chemical composition and distribution of Limonin in different parts of Melia azedarach fruits. Methods: 1 LC-MS/MS analysis conditions: LC separation of 17 analytes on a Chemometric Kinetex column (XB-MS/MS) The mobile phase consists of acetonitrile (A) and 0.1% (v/v) formic acid (B) at a flow rate of 300 muL/min. The following gradient procedures are used for quantitative quantification: 0.0-1.0 min, 45-55% A; 1.0-4.0 min, 55-60% A; 4.0-5.0 min, 60-50 min, 60-95% A; 5.0-0-10.0 min, 95% A; 10.0-10-10-0-95% A; 10.0-10-0-0-11.0-11.0, 95-45.0, 95-45-45.95-45.0-45% A; 1.0-0-1-0-4-4.0-4-4.0-4-4-4.0 positive ion mode Gas curtain gas, atomized gas (Gas 1) and heater gas (Gas 2) are set at 30, 50 and 55 psi, respectively. Nitrogen is used in all cases. The default MRM algorithm is used to monitor 34 ion pairs (2 ion pairs per compound) in the 90 s MR detection window and 0.7 s target scan time. In the enhanced product ion (EPI) mode, the information dependent acquisition (IDA) scan triggers the associated MS/MS mass spectrometry scans to better identify the object under test. The collision energy (CE) produces EPI mass spectrometry at a scan speed of 4000 Da/s with a 15 e V extended collision energy. In IDA settings, the precursor ions with the highest response signal (mass difference range 250 m Da) are selected, and the dynamic background is deducted for each detection scan. The signal threshold is 100 cps. EPI is identified by Master View? Residence time is the only difference between conventional MRM and preset MRM. It replaces the monitoring time window for each pair of ions with 20 ms. 2 The preset MRM LC-MS/MS method was validated, including linearity, minimum detection limit, precision, repeatability, stability, recovery and matrix effect. Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) are used to analyze LC-MS/MS data. Chemometric models can be used to visualize and interpret neem easily. Results: 1 In this study, a chemometric-assisted LC-MS/MS method was developed and validated for simultaneous determination of 17 limonins in different parts of Melia azedarach fruit. Compared with conventional MRM, preset MRM improved sensitivity and accuracy. Preset MRM LC-MS/MS method was well validated, including linear (R0.99), minimum detection. The optimal ultrasonic-assisted extraction conditions obtained by RSM were more accurate than those obtained by single factor analysis in 35 batches of neem fruits. All 17 kinds of Limonin showed distinct order, seed and fruit; 13 kinds of Limonin showed another order, seed peel and seed coat. Potential chemical markers for different parts of neem fruit were identified by PCA and OPLS-DA analysis based on LC-MS/MS data. Chemometric-assisted LC-MS/MS method was developed and validated for the simultaneous determination of 17 kinds of Limonin in different parts of Melia azedarach fruit. In addition, this study showed that LC-MS/MS was a multi-component quantitative development, optimization and data analysis, and a chemometric-assisted synthesis consisting of RSM, PCA and OPLS-DA. In addition, this study provides evidence for identifying and controlling the chemical composition and distribution of the fruit of Melia azedarach as a folk drug. Part III Preliminary experimental study on the antidepressant activity of Melia azedarach limonin Objective: To study the effect of Melia azedarach limonin on acute stress-induced depression in mice. Potential antidepressant activities and mechanisms. METHODS: The antidepressant activities of xylocarpin H, cipadesin A and xyloccensin K in Melia azedarach were studied by behavioral experiments, and their potential mechanisms were studied by neuroendocrine system. The antidepressant activities of xylocarpin H, cipadesin A and xyloccensin K in Melia azedarach (5,15,50). The effects of xylocarpin H and cipadesin A on the hypothalamus-pituitary-adrenal axis were evaluated by enzyme immunoassay. Results: Melia azedarach limonin significantly decreased the forced swimming in mice. The floating immobility time (xylocarpin H, cipadesin A and xyloccensin K, 15-50 mg/kg) was significantly decreased in mice tail suspension test (xylocarpin H and cipadesin A, 15-50 mg/kg, xyloccensin K, 5-50 mg/kg). Azadirachtin significantly increased the residence time (xylo) in the central area of the open field in mice tail suspension test. Carpin H was 5-50 mg/kg, cipadesin A was 15-50 mg/kg, xyloccensin K was 50 mg/kg, and did not affect spontaneous activity. In addition, repeated administration of xylocarpin H and cipadesin A significantly reduced serum corticosterone CORT (xylocarpin H was 15-50 mg/kg, cipadesin A was 5-50 mg/kg) and ACTH (adrenocorticotrophic hormone ACTH) levels in forced swimming mice. The levels of xylocarpin H and cipadesin A were 15-50 mg/kg, but had no effect on mice without stress treatment. CONCLUSION: Melia azedarach limonin xylocarpin H, cipadesin A and xyloccensin K have potential antidepressant activity in mice with acute stress-induced depression. The potential mechanism may be through inhibiting hypothalamus-pituitary. - The activity of the adrenal axis in response to stress. These data support further studies on the development of Limonin as a potential antidepressant and anxiety drug.
【学位授予单位】：河北医科大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R285.5
[Abstract]:Natural products play a leading role in the discovery of lead compounds for the development of drugs for the treatment of human diseases. In fact, the pharmacological active ingredients of plants and microorganisms are important approaches for the development of new drugs. Rapid and sensitive mass spectrometric techniques have been widely used in natural products. Mass spectrometric pyrolysis is helpful to identify and analyze the structures of compounds quickly, and repeat the known compounds to explore the unknown compounds. Mass spectrometry is rarely reported. Therefore, this study used mass spectrometry to identify and quantify the limonin components of Melia azedarach, and carried out a preliminary experimental study on the antidepressant activity of Melia azedarach limonin. This study laid a foundation for the rapid discovery of new compounds from Melia azedarach and the identification and quality control of Melia azedarach. The first part is the mass spectrometric identification of limonin from Melia azedarach. Objective: By analyzing the secondary mass spectrometry of limonin from Melia azedarach (Q-TOF-MS/MS), to explore its mass spectrometric cleavage pathway and summarize the cleavage rule. Methods: 1 UHPLC-Q-TOF-MS/MS analysis conditions: Kinetex-C18 column (100 m m *2.1 m m, 2.6 micron), column temperature: 40 C. Mobile phase: acetonitrile A, water-0.1% formic acid-1 M ammonium acetate solution. Gradient elution procedure: 10% A (0-5 min); % A (5-10 min); 30-35% A (10-20 min); 35-50% A (20-35 min); 50-95% A (35-45 min); 95% A (45-50 min). Flow rate: 0.4 m L/min. Sample volume: 2 mu L. Duo Spray Electrospray Ion Source (ESI); Turbospray temperature: 600 (?) CUR: 35 psi; Gas 1: 50 psi; Heating gas (50 psi); Spray: 50 psi in positive ion mode The voltage (ion spray voltage) is 5500 V and the unwrapping voltage (DP) is 80 V. Information correlation acquisition mode includes TOF-MS probe scan 100-1000 Da (200 ms) and up to 8 dependent TOF-MS/MS scan 50-1000 Da (100 ms), collision energy 35 e V, collision spread energy (+15 e V.2) identification strategy: empirical molecular formula calculation and debris ion prediction enabled The known compounds were identified by comparing retention time, accurate mass and MS/MS mass spectrometry with the reference materials. Based on the mass spectrometric pyrolysis law, accurate mass, retention time and phytochemical literature, the known compounds were identified and the unknown compounds were speculated. Mass spectrometric fragmentation of Limonin in Melia azedarach fruit. These fragmentation patterns mainly include loss of acyl or acyloxy side chains, loss of water and CO molecules. 2 UHPLC-Q-TOF-MS/MS was used to identify limonin compounds in Melia azedarach fruit. At least 300 compounds were detected from the extracts of Melia azedarach fruit. The results were compared with those of the control. Seventeen known compounds were identified. Seven potential new compounds were identified. CONCLUSION: 1. Mass spectrometric fragmentation of 17 limonins from Melia azedarach was systematically studied for the first time. 2 U-HPLC-Q-TOF-MS/MS methods were used to identify the limonins from Melia azedarach. The identification of limonoids in the fruit of Melia azedarach by UHPLC-Q-TOF-MS/MS is simple, rapid, sensitive and specific. The identification of these compounds provides an important basis for further study on the phytochemistry, metabolites identification and quality control of the limonoids in Melia azedarach. Objective: To develop and validate a chemometric-assisted LC-MS/MS method for the determination of 17 kinds of Limonin in Melia azedarach fruits, and to reveal the chemical composition and distribution of Limonin in different parts of Melia azedarach fruits. Methods: 1 LC-MS/MS analysis conditions: LC separation of 17 analytes on a Chemometric Kinetex column (XB-MS/MS) The mobile phase consists of acetonitrile (A) and 0.1% (v/v) formic acid (B) at a flow rate of 300 muL/min. The following gradient procedures are used for quantitative quantification: 0.0-1.0 min, 45-55% A; 1.0-4.0 min, 55-60% A; 4.0-5.0 min, 60-50 min, 60-95% A; 5.0-0-10.0 min, 95% A; 10.0-10-10-0-95% A; 10.0-10-0-0-11.0-11.0, 95-45.0, 95-45-45.95-45.0-45% A; 1.0-0-1-0-4-4.0-4-4.0-4-4-4.0 positive ion mode Gas curtain gas, atomized gas (Gas 1) and heater gas (Gas 2) are set at 30, 50 and 55 psi, respectively. Nitrogen is used in all cases. The default MRM algorithm is used to monitor 34 ion pairs (2 ion pairs per compound) in the 90 s MR detection window and 0.7 s target scan time. In the enhanced product ion (EPI) mode, the information dependent acquisition (IDA) scan triggers the associated MS/MS mass spectrometry scans to better identify the object under test. The collision energy (CE) produces EPI mass spectrometry at a scan speed of 4000 Da/s with a 15 e V extended collision energy. In IDA settings, the precursor ions with the highest response signal (mass difference range 250 m Da) are selected, and the dynamic background is deducted for each detection scan. The signal threshold is 100 cps. EPI is identified by Master View? Residence time is the only difference between conventional MRM and preset MRM. It replaces the monitoring time window for each pair of ions with 20 ms. 2 The preset MRM LC-MS/MS method was validated, including linearity, minimum detection limit, precision, repeatability, stability, recovery and matrix effect. Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) are used to analyze LC-MS/MS data. Chemometric models can be used to visualize and interpret neem easily. Results: 1 In this study, a chemometric-assisted LC-MS/MS method was developed and validated for simultaneous determination of 17 limonins in different parts of Melia azedarach fruit. Compared with conventional MRM, preset MRM improved sensitivity and accuracy. Preset MRM LC-MS/MS method was well validated, including linear (R0.99), minimum detection. The optimal ultrasonic-assisted extraction conditions obtained by RSM were more accurate than those obtained by single factor analysis in 35 batches of neem fruits. All 17 kinds of Limonin showed distinct order, seed and fruit; 13 kinds of Limonin showed another order, seed peel and seed coat. Potential chemical markers for different parts of neem fruit were identified by PCA and OPLS-DA analysis based on LC-MS/MS data. Chemometric-assisted LC-MS/MS method was developed and validated for the simultaneous determination of 17 kinds of Limonin in different parts of Melia azedarach fruit. In addition, this study showed that LC-MS/MS was a multi-component quantitative development, optimization and data analysis, and a chemometric-assisted synthesis consisting of RSM, PCA and OPLS-DA. In addition, this study provides evidence for identifying and controlling the chemical composition and distribution of the fruit of Melia azedarach as a folk drug. Part III Preliminary experimental study on the antidepressant activity of Melia azedarach limonin Objective: To study the effect of Melia azedarach limonin on acute stress-induced depression in mice. Potential antidepressant activities and mechanisms. METHODS: The antidepressant activities of xylocarpin H, cipadesin A and xyloccensin K in Melia azedarach were studied by behavioral experiments, and their potential mechanisms were studied by neuroendocrine system. The antidepressant activities of xylocarpin H, cipadesin A and xyloccensin K in Melia azedarach (5,15,50). The effects of xylocarpin H and cipadesin A on the hypothalamus-pituitary-adrenal axis were evaluated by enzyme immunoassay. Results: Melia azedarach limonin significantly decreased the forced swimming in mice. The floating immobility time (xylocarpin H, cipadesin A and xyloccensin K, 15-50 mg/kg) was significantly decreased in mice tail suspension test (xylocarpin H and cipadesin A, 15-50 mg/kg, xyloccensin K, 5-50 mg/kg). Azadirachtin significantly increased the residence time (xylo) in the central area of the open field in mice tail suspension test. Carpin H was 5-50 mg/kg, cipadesin A was 15-50 mg/kg, xyloccensin K was 50 mg/kg, and did not affect spontaneous activity. In addition, repeated administration of xylocarpin H and cipadesin A significantly reduced serum corticosterone CORT (xylocarpin H was 15-50 mg/kg, cipadesin A was 5-50 mg/kg) and ACTH (adrenocorticotrophic hormone ACTH) levels in forced swimming mice. The levels of xylocarpin H and cipadesin A were 15-50 mg/kg, but had no effect on mice without stress treatment. CONCLUSION: Melia azedarach limonin xylocarpin H, cipadesin A and xyloccensin K have potential antidepressant activity in mice with acute stress-induced depression. The potential mechanism may be through inhibiting hypothalamus-pituitary. - The activity of the adrenal axis in response to stress. These data support further studies on the development of Limonin as a potential antidepressant and anxiety drug.
【学位授予单位】：河北医科大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R285.5
【相似文献】
相关期刊论文 前10条
1 霍长虹;郭栋;沈立茹;王伟;张青;张Z诶，
本文编号：2203415
本文链接：https://www.wllwen.com/shoufeilunwen/yxlbs/2203415.html
教材专著
