蝉蜕提取物提取工艺优化和药效学研究

【作者】 李进京；

【导师】 沈德凤； 西冈丰； 宫村充彦； 孙广臣；

【作者基本信息】 佳木斯大学 ， 药物化学， 2009， 硕士

【摘要】 目的:研究蝉蜕提取物的最佳提取工艺和体外抗氧化活性;研究蝉蜕95%乙醇提取物(PC95%)的镇咳作用和对变应性接触性皮炎(ACD)的作用,为进一步开发蝉蜕的临床应用提供理论依据。方法:应用均匀设计U5(53)安排实验,以提取率为考察指标,对蝉蜕提取物进行提取工艺优化;分别用黄嘌呤氧化酶法和电子自旋共振波谱法(ESR)测定蝉蜕提取物体外抗氧化活性;用浓氨水引咳法和枸橼酸引咳法分别制造小鼠和豚鼠咳嗽模型,以实验动物咳嗽潜伏期和3min内咳嗽次数为观察指标,探究PC95%的镇咳作用;用二硝基氟苯(DNFB)反复涂抹法制造大鼠ACD模型,通过测定大鼠病变耳肿胀度、血清及耳组织中相关免疫细胞因子的变化,综合评价PC95%对ACD的作用。结果:蝉蜕提取物的最佳提取工艺是选择回流提取法,以水为提取溶剂,提取前不浸泡,料液比1:20,回流提取3次,每次3h;不同提取工艺所得提取物中,PC95%抗氧化活性最强。在动物实验中以PC95%为研究对象,PC95%可显著延长实验动物咳嗽潜伏期(P<0.05)和减少3min内咳嗽次数(P<0.05);PC95%能显著减轻大鼠病变耳组织肿胀度(P<0.05);PC95%能显著抑制由DNFB诱导的ACD过敏介质释放和炎症的发生。结论:以上实验结果提示:应用均匀设计U5(53)安排实验获得蝉蜕提取物的最佳提取工艺可为蝉蜕提取物工业化生产起指导作用;95%乙醇提取物体外抗氧化活性最强,该提取物有良好的镇咳作用,且能明显抑制由DNFB诱导的大鼠ACD过敏介质释放和炎症反应发生,减轻组织损伤,为进一步开发蝉蜕的临床应用提供了理论依据。更多还原

【Abstract】 Objective: To study the best extraction technology of Periostracum Cicadae (PC) and the anti-oxidation effect in vitro of PC; To study the antitussive effect in 95 percent ethnol extract from PC and the effect of allergic contact dermatitis (ACD). This research is to provide theoretical basis for the further development of PC in clinic application.Methods: The extraction technology of PC was optimized by U5(53) Uniform Design. Extrasomatic antioxidative activities were determined from PC by xanthine oxidase method and Electron Spin Resonance (ESR) separately.The animal model of cough was induced by ammonia water stronger and citric acid. The latency of cough and the times of coughing in 3 minutes were observed to evaluate the relieving cough effects of the extract of PC95;The animal model of ACD was induced by repeatedly challenging the ear of previously sensitized rats with dinitrofluorobenzene (DNFB). The thickness of the inflamed ear regions and associated immune cytokine in the ear tissues homogenate or in serum were measured to investigate the effect of PC95 on ACD animal model.Results: The best extraction technology from PC was refluxing method ,which was water as solvent , extraction time 3h and solid-liquid ratio 1:20,3 times ; The extract of 95% ethanol showed the greatest antioxidative activity comparied with extracts from different technology extracts. Therefore, in the animal experiment, the extract of 95% ethanol(PC95) was used as a studying object. The extract from PC95 was able to lengthen obviously latency of cough (P<0.05) and decrease the number of coughing in 3 minutes significantly (P<0.05).Besides, PC95 could decrease evidently swelling degree of ear tissue caused by pathological changes, significant differences were shown between PC95 group and model group(P<0.05). PC95 could also inhibit notably releasing of ACD allergic medium induced by DNFB and development of inflammation.Conclusion: According to experimental results above metioned ,we can conclude that the best extract technology from PC by U5(53) Uniform Design was able to guide industrial production; 95% ethanol showed the greatest antioxidative activity, better relieving cough effect , inhibiting notably releasing of ACD allergic medium induced by DNFB and development of inflammation,relieving tissue damage, this study has provided theoretical basis for the further development of PC in clinic application.更多还原