咖啡酸对硝基苯乙酯抗实验性血小板聚集及其药动学分析

发布时间：2018-05-15 21:26

本文选题：咖啡酸对硝基苯乙酯 + 血小板　；参考：《西南大学》2014年硕士论文

【摘要】：咖啡酸苯乙酯来源于蜂胶,是其主要活性成分之一,文献报道它具有多种生物活性,例如抗病毒、抗菌、抗氧化、免疫调节、抗肿瘤、心肌缺血再灌注损伤保护等生理活性,它在民间的用药史很悠久,主要用于中东地区,在民间蜂胶是一种常用的养生保肝的天然产物；咖啡酸苯乙酯的分子结构为O-二羟基苯基结构和两个邻位的酚羟基,前面的基团是清除自由基的活性基团,而后者是容易被氧化的。本文研究了咖啡酸苯乙酯的衍生物,考察其在抗血小聚集方面的活性及其可能的机制和药动学研究。本文研究了咖啡酸对硝基苯乙酯抑制体外大鼠的血小板聚集的活性,分别比较了空白对照组、阳性对照组(咖啡酸苯乙酯)、实验组的(咖啡酸对硝基苯乙酯)抗血小板聚集的活性,研究了他们对胶原激活的血小板的生理代谢过程的影响,如血小板聚集率的改变,为进一步阐述咖啡酸对硝基苯乙酯抑制胶原诱导的血小板聚集的机制,本文考察了其对血小板处理后,血小板内的一氧化氮的释放、环磷酸鸟苷(cGMP)变化、血栓素A2(TXA2)的形成、环氧合酶-1(COX-1)的活性、5-羟色胺(5-HT)的分泌释放情况。这些物质的变化都与血小板的激活、信号的传导、代谢有着密切的联系。体外的实验证明,咖啡酸对硝基苯乙酯和咖啡酸苯乙酯均能够抑制胶原(Collagen)诱导的大鼠洗涤血小板和PRP血浆的血小板的聚集,咖啡酸对硝基苯乙酯(CAPE-NO2)在1.5-12μM对血小板的聚集抑制呈现剂量依赖性,抑制了血小板的聚集率分别为62%、52%、46%、34%,与空白组相比在咖啡酸对硝基苯乙酯12gM时对血小板的聚集抑制率为55%,而咖啡酸苯乙酯(1.5μM)对血小板的聚集抑制率是14%,因此咖啡酸苯乙酯和咖啡酸对硝基苯乙酯对血小板有很好的抑制聚集活性。血小板中的NO水平显著的被咖啡酸对硝基苯乙酯和咖啡酸苯乙酯升高,CAPE-NO2对血小板中的NO的作用呈剂量依赖性；咖啡酸对硝基苯乙酯显著的增加了细胞的cGMP的含量,与剂量成正比关系。咖啡酸对硝基苯乙酯在浓度为1.5、3、6、12μM时抑制了血栓素B2的产生分别是9.5%、23%、40%、51%,但是咖啡酸苯乙酯在1.5μM时的抑制率为18%。咖啡酸对硝基苯乙酯在浓度1.5-12gM时抑制环氧合酶-1的活性；在不同浓度(1.5、3、6、12μM)下的5-HT的释放量为454μg/L、435μg/L、408μg/L、376μg/L,咖啡酸对硝基苯乙酯在12μM时对5-HT的抑制率为87%,而1.5μM的咖啡酸苯乙酯为19%。它的抑制作用的有可能涉及了以下的机制：咖啡酸对硝基苯乙酯激活了NO/cGMP的信号通路,分别增加了NO的释放过程和cGMP的产生,咖啡酸对硝基苯乙酯也抑制了环氧合酶-1的活性,导致血栓素B2的减少；最后咖啡酸对硝基苯乙酯减少了5-羟色胺从激活的血小板的分泌。通过采用HPLC-UV建立了咖啡酸对硝基苯乙酯肌肉注射大鼠血浆中的含量测定方法,并进行药代动力学研究。色谱柱为SepaxHP-C18(4.6mm x250mm,5μm),流动相为乙腈-水(55：45),检测波长329nm,流速为1mL·min-1,柱温30℃,进样量20μL,血液中咖啡酸对硝基苯乙酯的浓度在50-2000ng.mL-1范围内线性关系良好,相关系数r=0.9999,定量下限为0.05μg·mL-1。用DAS3.0软件分析后,咖啡酸对硝基苯乙酯在SD大鼠体内的药物代谢动力学符合二室开放模型,Cmax为976.66μg·L-1, tmax为1h,tl/2α为0.95h,t1/2β为5.51h, AUC0-t为2649.77μg·h·L-1, AUC0-∞为2837.77μg·h·L-1.
[Abstract]:Caffeic acid benzoate is one of the main active ingredients of propolis. It is reported that it has many biological activities, such as antiviral, antibacterial, antioxidation, immunomodulating, anti tumor, protection of myocardial ischemia reperfusion injury and so on. It has a long history in the folk medicine, mainly used in the Middle East, and is a common use in folk propolis. The natural product of health preserving liver preservation; the molecular structure of caffeic acid benzoethyl ester is O- two hydroxy phenyl structure and two ortho phenolic hydroxyl groups. The front group is the active group for scavenging free radicals, and the latter is easily oxidized. The mechanism of energy and the study of pharmacokinetics.
This paper studied the activity of caffeic acid on the inhibition of platelet aggregation in rats with nitrobenzene, compared with the blank control group, the positive control group (caffeic acid benzol), the activity of the antiplatelet aggregation in the experimental group (caffeic acid to nitrobenzene ethyl), and the effect of their effects on the physiological metabolism of the platelets activated by collagen. In order to further elaborate the mechanism of platelet aggregation induced by caffeic acid against nitrobenzene, the release of nitric oxide in platelets, the changes in cGMP, the formation of thromboxane A2 (TXA2), the activity of cyclooxygenase -1 (COX-1), 5- hydroxytryptamine (5-), and 5- hydroxytryptamine (5-) were investigated. The secretion and release of HT. These changes are closely related to platelet activation, signal transduction and metabolism.
In vitro experiments showed that caffeic acid against nitrobenzene and caffeic acid benzoethyl could inhibit the aggregation of platelet in rat washed platelets and PRP plasma induced by collagen (Collagen). The inhibition of platelet aggregation by caffeic acid to nitrobenzene (CAPE-NO2) was dose-dependent on the inhibition of platelet aggregation in 1.5-12 mu M, inhibiting the aggregation rate of platelets respectively 62%, 52%, 46%, 34%, compared with the blank group, the inhibition rate of platelet aggregation was 55% on the caffeic acid to nitrobenzene ethyl 12gM, while the inhibition rate of caffeic acid benzoate (1.5 mu M) on platelet aggregation was 14%. Therefore, caffeic acid benzoethyl and caffeic acid had a good inhibitory aggregation activity on the platelets. The NO level in platelets The effect of caffeic acid on nitrobenzene and caffeic acid benzoate was significantly increased, and the effect of CAPE-NO2 on the NO in platelets was dose-dependent; caffeic acid to nitrobenzene significantly increased the cGMP content of the cell, and was proportional to the dose. Caffeic acid against nitrobenzene at the concentration of 1.5,3,6,12 u M inhibited the production of thromboxane B2. The results were 9.5%, 23%, 40%, 51% respectively, but the inhibition rate of caffeic acid benzol at 1.5 um M was 18%. caffeic acid to nitrobenzene at the concentration of 1.5-12gM, the activity of cyclooxygenase -1 was inhibited. The release of 5-HT at different concentrations (1.5,3,6,12 mu M) was 454 mu, 435, 408, 408 mu, 376 mu g/L, and caffeic acid to nitrobenzene was in 12 mu M. The inhibitory rate was 87%, while the inhibitory action of 1.5 M caffeic acid benzol to 19%. may involve the following mechanism: caffeic acid activates the NO/cGMP signaling pathway to nitrobenzene, which increases the release process of NO and the production of cGMP, and the caffeic acid against nitrobenzene also inhibits the activity of cyclooxygenase -1, leading to thromboxane. The reduction of B2; finally, caffeic acid on NP reduced the secretion of 5- HT from activated platelets.
The content determination method of caffeic acid to nitrobenzene was established by HPLC-UV, and the pharmacokinetics was studied. The chromatographic column was SepaxHP-C18 (4.6mm x250mm, 5 mu m), the mobile phase was acetonitrile water (55:45), the detection wavelength 329nm, the velocity of 1mL. Min-1, the column temperature of 30, 20 mu L, and the caffeic acid to nitrate in the blood. The relationship between the concentration of ethyl benzene ethyl ester in the range of 50-2000ng.mL-1 is good, the correlation coefficient is r=0.9999, the quantitative lower limit is 0.05 mu g. ML-1. is analyzed by DAS3.0 software. The pharmacokinetics of caffeic acid to nitrobenzene ethyl ester in SD rats conforms to the two chamber open model, Cmax is 976.66 Mu G. L-1, Tmax is 1H, tl/2 alpha is UC0-t is 2649.77 g h L-1 and AUC0- infinity is 2837.77 g g h L-1..

【学位授予单位】：西南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R96

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