The spectrum-effect relationship study on Tripterygium Wilfordii glycoside tablet based on therapeutic effect and toxicity

雷公藤多苷片是由雷公藤的干燥根提取、纯化制得的，具有良好的抗炎、免疫抑制的药理作用，临床上广泛用于治疗类风湿关节炎、肾病综合征等疾病，疗效确切。但临床不良反应高发，且临床应用表明不同厂家的雷公藤多苷片药效和毒副反应有较大差异。由于雷公藤化学成分复杂，多个成分既为有效成分又为毒性成分，而目前多仅对雷公藤甲素(雷公藤内酯醇)或雷公藤内酯甲进行含量控制，无法全面的对雷公藤多苷片的药效和毒性进行控制。明确药效成分和毒性成分，建立全面科学、符合中药制剂整体起效特点的质量控制方法，是当前研究的热点和难点。本文拟通过体外实验，评价不同厂家的药效及毒效差异，结合雷公藤多苷片高效液相指纹图谱，通过统计学方法建立...Tripterygium wilfordii glycoside (TWG) tablet which extracted from the traditional Chinese herb Tripterygium wilfordii (TWHF) had good anti-inflammatory and immunesuppres¬sive therapeutic effect, which was widely used to treat the rheumatoid arthritis, nephrotic syndrome and other diseases in clinical. But the different quality and high incident rate of side effects inhibited its clinical app...学位：医学硕士院系专业：医学院_药理学学号：2452012115321

雷公藤减毒增效工艺的研究进展

目的:为更安全使用雷公藤提供参考。方法:检索中国知网,从制备工艺上对雷公藤减毒增效的技术手段进行文献综述。结果:雷公藤减毒增效的制备工艺主要有传统的炮制减毒、超临界流体萃取技术和加压溶剂提取技术。制剂新技术制备的如胃漂浮缓释片、微乳凝胶等均能明显缓解患者的不良反应,减毒效果明显。结论:不同的制备方法可为研究更加安全有效的雷公藤制剂提供理论依据。福建省科技计划项目(No.2012I1001

Spectrum-effect relationship of immunosuppressive activity of Leigongteng Duogan Tablet based on partial least squares regression analysis

目的初步探讨雷公藤多苷片免疫抑制作用及其HPlC指纹图谱间的相关性。方法采用HPlC建立7个厂家生产的雷公藤多苷片醋酸乙酯提取物的指纹图谱,选择刀豆蛋白A(COn A)刺激的小鼠脾淋巴细胞模型进行免疫抑制药效学研究,分别考察雷公藤多苷片对小鼠脾淋巴细胞增殖以及细胞因子γ干扰素(Ifn-γ)分泌的影响;采用偏最小二乘回归分析法分析谱效关系。结果建立了7个厂家生产的雷公藤多苷片的HPlC指纹图谱,从HPlC指纹图谱中提取出18个能够表征差异的特征峰。药效研究结果显示不同厂家生产的雷公藤多苷片均可显著抑制COn A诱导的小鼠脾淋巴细胞增殖和Ifn-γ的分泌(P<0.05)。偏最小二乘回归法分析结果显示,以抑制小鼠脾淋巴细胞增殖IC50值为药效指标时第1、2、6、7、17、18号峰为药效峰;以Ifn-γ分泌抑制率为指标时,第1、2、6、7、8、9、12、13号峰为药效峰。结论第1、2、6、7、8、9、12、13、17、18号峰所代表的化合物可能为雷公藤多苷片中发挥免疫抑制主要有效成分。Objective To investigate the immunosuppressive components in Leigongteng Duogan Tablet(LDT).Methods High performance liquid chromatography(HPLC) was applied to establish the fingerprint for LDT from seven manufactures.The immunosuppressive effect of LDT was observed on Con A-induced mice spleen cells, and the cell proliferation and contents of IFN-γ were recorded.Then the partial least squares regression(PLSR) analysis was used to investigate its spectrum-effect relationship.Results The HPLC fingerprint for LDT from seven manufactures was established, and 18 characteristic peaks were confirmed.The in vitro cell experimental results showed that the proliferation of mice spleen cells and secretion of IFN-γ were inhibited by all LDT significantly compared with the control group.According to the results of PLSR, No.1, 2, 6, 7, 17, and 18 peaks were strongly related to the proliferation inhibition of mice spleen cells and No.1, 2, 6, 7, 8, 9, 12, and 13 peaks were related to the secretion of IFN-γ.Conclusion The No.1, 2, 6, 7, 8, 9, 12, 13, 17, and 18 peaks are the principal immunosuppressive substances in LDT.福建省科技计划重大项目(2012I1001

Fingerprint and spectrum-effect relationships on Tripterygium glycosides preparation

采用高效液相色谱法(HPlC)建立7个厂家雷公藤多苷片醋酸乙酯提取物的指纹图谱,得到11个共有峰,各厂家指纹峰种类和含量差异较大。选择COn A刺激的小鼠脾淋巴细胞模型进行药效学研究,分别给予30,15,7.5,3.75,1.88,0.94Mg·l~(-1)的雷公藤多苷片提取物进行处理,48 H后测定各药物对小鼠脾淋巴细胞增殖的作用,发现各厂家的雷公藤多苷片均对小鼠脾淋巴细胞增殖有显著抑制作用(P10>8>7>6>11>4>9>2>3>1。该文初步探讨了雷公藤的物质基础,为明确雷公藤的作用机制,建立更加全面可靠的质量控制方法提供理论依据与数据支持。Tripterygium glycosides preparation which extracted from the traditional Chinese herb Tripterygium wilfordii(TWHF),was widely used to treat the autoimmune diseases.Previous works demonstrated that TWHF had potent anti-inflammatory and immunosuppressive properties.But the different quality and high incident rate of side effects of different manufactures inhibited its clinical application.Since TWHF had been generally known to play a therapeutical effect by synergism of multiple constituents,it was necessary to build the relationship between the HPLC fingerprint and bioactivity so as to ensure the quality safety and efficacy.The HPLC fingerprint showed that description and content of peaks from different manufactures were diverse.Only 11 common peaks were found.In this study,mice spleen cells stimulated by Con A were used to test the proliferation inhibition bioactivity of TWHF preparations,which were incubated with 30,15,7.5,3.75,1.88 and 0.94 mg·L~(-1)TWHF preparations for 48 h.The results showed that mice spleen cells proliferation was inhibited by all TWHF preparations significantly compared with the control group,which suggested the TWHF preparations showed immune suppress activity.The TWHF preparations from 7 manufacture showed different IC_(50)value,which might belong to different contents which showed in the HPLC fingerprint.Moreover,a relationship between the HPLC fingerprint and the bioactivity were established to identify important constituents by grey relational analysis(GRA).The result showed that all the contents were relative with the IC_(50),especially No.5 and 10 peaks,but No.1 peak,which was proved to be triptolide,had few contribute to the inhibition of mice spleen cells proliferation.The study of relationship between the HPLC fingerprint and the IC_(50) by GRA could help to investigate mechanism of bioactive and provide an evidence for the quantification of multi-constituents.福建省科技计划重大项目(201211001

门静脉和下腔静脉阻断与开放后门脉高压模型犬肺组织和肺动脉的病变特征

【目的】 观察门静脉和下腔静脉阻断与开放后门脉高压模型犬肺组织与肺动脉的病理学特点，从病理学角度初步探讨终末期肝硬化肝移植围术期急性肺高压和肺损伤的发生机制。【方法】 正常家犬18只，随机均分为阴性对照组（仅用于取肺组织和肺动脉标本）、对照组和门脉高压模型组（采用部分结扎门静脉的方法建立门脉高压症犬的模型并饲养12周），进行门静脉和肝后下腔静脉阻断与开放实验，在术毕后取右下肺动脉和肺组织，观察其病理形态学特征。【结果】 模型组与对照组肺组织在经历门静脉和肝后下腔静脉阻断与开放后均存在不同程度病理形态学改变，但以模型组改变更为显著。门脉高压症犬门脉高压犬肺小动脉密度、肺小动脉中膜厚度和血管壁面积/血管总面积（WA/TA）均显著大于阴性对照组和对照组；模型组的肺血管发生了以中膜增厚、动脉管壁肥厚为主要特征的血管重构现象。【结论】 模型组犬易发生急性肺高压及肺损伤的特殊的病理学改变与存在门脉高压密切相关