表儿茶素镇痛抗炎作用与机理研究

1分类号 学号学校代码 密级硕 士 学 位 论 文表儿茶素镇痛抗炎作用与机理研究Study on the anti inflammatory effect and mechanism of the effect of the anti-inflammatory action of the tea研 究 生：2表儿茶素镇痛抗炎作用与机理研究指导教师：学科专业：研究方向：培养学院：3独创性声明本人声明所呈交的论文是我个人在导师指导下进行的研究工作及取得的研究成果。尽我所知，除了文中特别加以标注和致谢的地方外，论文中不包含其他人已经发表或撰写过的研究成果，也不包含为大学或其它教育机构的学位或证书而使用过的材料。与我一同工作的同志对本研究所做的任何贡献均已在论文中作了明确的说明并表示了谢意。研究生签名：日期： 年 月 日关于论文使用授权的说明本人完全了解有关保留、使用学位论文的规定，即：学校有权保留送交论文的复印件和磁盘，允许论文被查阅和借阅，可以采用影印、缩印或扫描等复制手段保存、汇编学位论文。同意可以用不同方式在不同刊物上发表，传播学位论文的全部或部分内容。(保密的学位论文在解密后应遵守此协议)研究生签名： 导师签名：日期： 年 月 日 日期： 年 月4表儿茶素镇痛抗炎作用与机理研究目录目录 5Abstract .9第一章前言 .121.1 炎症和疼痛的发生机制 .121.2 表儿茶素的研究进展 .141.2.1 化学性质 .141.2.2 药理作用 .141.2.3 临床应用 .151.3 本研究的目的和意义 .181.3.1 体内镇痛抗炎作用与机理研究 .181.3.2 体外抗炎作用与机理研究 .18第二章表儿茶素阵痛作用于机制研究 .192.1 材料与方法 .192.1.1 实验动物 .192.1.2 试剂与仪器 .192.1.3 扭体实验 .192.1.4 热板实验 .192.1.5 福尔马林实验 .20552.2 结果与分析 .202.2.1 表儿茶素对小鼠冰醋酸致痛实验的影响 .202.2.2 表儿茶素对小鼠热板法实验的影响 .222.3 结论与讨论 .26第三章表儿茶素抗炎作用于机制研究 .283.1 材料与方法 .283.1.1 供试菌株 .283.1.2 试剂与仪器 .283.1.3 NO 检测 .283.1.4 台盼蓝细胞活性检测 .303.1.5 WB 检测 .333.1.6 ELISA 实验 .353.1.7 MTT 实验 .373.2 结果与分析 .373.2.2 细胞增殖实验结果 .393.2.3 WB 实验结果 .413.2.4 台盼蓝细胞染色结果 .433.3 结果与分析 .43第四章结论 .466表儿茶素镇痛抗炎作用与机理研究参考文献 .47文献综述 .52摘要目的：通过对小鼠的实验研究，研究表儿茶素的镇痛抗炎作用与作用机制。77方法：采用醋酸扭体实验、热板实验、福尔马林实验、角叉菜胶诱导足肿胀实验等动物疼痛模型，以阐明表儿茶素的镇痛抗炎作用；采用酶联免疫吸附法测定冰醋酸致痛小鼠血清中 NO、白细胞介素-1（IL-1 ），细胞因子- （TNF- ）、前列素E2（PGE2）的含量，以阐明表儿茶素的镇痛抗炎作用机制。采用培养巨噬细胞RAW2647，采用 Griess 显色法和 PGE2 酶联免疫法分别测定表儿茶素对脂多糖(LPS) 诱导细胞释放 NO 和 PGE2 等炎症因子的抑制率，并采用 MTT 法测定药物对细胞活力的影响程度。结果：（1）在扭体次数及扭体抑制率方面，与空白对照组相比，表儿茶素高、中、低 3 个剂量组均可显著减少小鼠扭体次数，最大抑制率达 68.40%，呈现剂量依赖性，差异显著，有统计学意义(P0.05)。在第二时相累积反应时间降低方面，布洛芬缓释胶囊组、元胡止痛片剂量组、表儿茶素高、中剂量组均优于表儿茶素低剂量组，差异显著，有统计学意义(P0.05)。研究表明表儿茶素对热刺激引起的小鼠热浴甩尾有一定的镇痛作用，但只有高剂量组作用效果稍强于阳性对照药物元胡止痛片组。（4）LPS 刺激能够是小鼠体内的一氧化氮含量升高，而表儿茶素能够降低小鼠体内的一氧化氮含量，而且剂量越高，一氧化氮含量降低的越低。（5）空白组中 NO 浓度仅为 5.23mol/L，在 1g/mL 的 LPS 刺激8表儿茶素镇痛抗炎作用与机理研究下，巨噬细胞释放出大量的 NO(47.56mol/L)，两组间比较，差异显著，有统计学意义（P 0.05). In the second phase accumulation response time reduction, Ibuprofen Sustained release capsule group, Rhizoma Corydalis analgesic dose group, table catechin high and middle dose group were superior to the table catechin low dose group, significant difference and was statistically significant (P 0.05). Studies show that catechin on thermal stimulation in mice induced by hot bath flick analgesic effect, but only high dosage group. The effect was stronger in the positive control drug Corydalis analgesic tablets group. (4) LPS can increase the content of nitric oxide in mice, but it can decrease the content of nitric oxide in mice. (5) NO concentration in the blank group was only 5.23 mol/L, and the release of NO (47.56 mol/L), two LPS (1 g/mL), the difference was statistically significant (P0.01). The results showed that LPS could induce the release of a large number of inflammatory cells in RAW 264.7 cells, and the inflammatory model was successful.Conclusion: (1) table catechin high and middle dose group can reduce the acetic acid induced writhing response times of mice with pain, reduce formalin induced pain mice of the first phase and second phase accumulation reaction time, improve the method of hot plate induced pain in mice mean pain threshold and pain threshold rate. Epicatechin in high dose group can improve the mice pain caused by hot bath tail flick method the average pain threshold and pain threshold rate. In the four induced pain model experiments, the difference between high dose and medium dose group and blank control group were significantly different, and it was dose dependent. A series of experimental studies on the pharmacological effects of analgesia showed that the pharmacological effects of central analgesia or peripheral analgesia. (2) high, middle and low dose groups could reduce the MDA content in serum of mice induced by acetic acid and enhance the activity of SOD in serum of mice with acetic acid induced pain, and the analgesic effect of the mice was related to the inhibition of oxygen free radicals. (3) the expression of c-fos protein and GFAP protein in brain tissue of brain tissue of mice was significantly lower than that of model control group. The expression of c-fos protein and GFAP protein as the index of pain response, the stronger the body pain, so the expression of c-fos protein and GFAP protein was down regulated, and the expression of c-fos protein and GFAP protein was related. The results showed that the inhibition of the nociceptive pain information was inhibited by