基于差异蛋白组学技术研究红、黑蛤蚧的谱效关系

Gekko gecko has the function of nourishing lung and stomach which was used as a traditional medicine to treat many diseases, such as shortness of breath,hemoptysis, impotence, seminal emission and so on. For a long time, The red gecko whether has the black gecko effect is similar with the existence of different views. In view of the fact that the wild population of Guangxi black Tokay plummeted in recent years, red point from Southeast Asia is currently the main varieties of gecko medicine circulation on the market, It is very necessary that scientific Evaluation on the Pharmacodynamics of the red and black gecko,especially red gecko.In this project, we will utilize the protein chip combined with Surface-enhanced laser desorption ionization/time-of-flight mass spectrometry (SELDI-TOF-MS) technology to compare the protein expression differences among all Tokay samples entirely, which include the different parts of Tokays from different genera, genus and sex. After analysis of the data, we aim to find some pesticide effect-related proteomes with high expression or low expression levels and construct a protein expression fingerprint which is related to the pesticide effect of Tokay. Through study on treatment of asthma about the red tokay and black tokay and protein expression fingerprint establish a spectrum-activity relationship.The relationship will be a useful tool to estimate the medicine value of Black Tokay and Red Tokay in the future studies.

蛤蚧(Gekko gecko)具有补肺滋胃，定喘止咳的功能,用于治疗虚喘气促，劳嗽咳血，阳痿遗精等症。长期以来，红点蛤蚧是否具有与黑点蛤蚧相似的药效存在着不同的看法。鉴于近年来广西黑点蛤蚧野生数量的骤减，产自东南亚地区的红点蛤蚧是目前蛤蚧药材市场上的主要流通品种，对红、黑点蛤蚧的药效，特别是红蛤蚧的药效进行科学评估显得十分必要。利用蛋白组学的蛋白芯片技术，用表面增强激光飞行时间质谱技术（SELDI-TOF-MS）技术，打破传统的无法全面比较样本全部蛋白差异的局限性，对不同种属，不同性别的蛤蚧以及蛤蚧不同部位的蛋白表达谱进行研究，通过相应的软件分析，找到跟药效相关的特定的多个高表达蛋白和低表达蛋白组，根据这些数据建立与药效具有相关性的蛋白表达指纹图谱，并通过对红、黑蛤蚧平喘作用研究与蛋白表达指纹图谱建立的“谱效”关系来评估黑点蛤蚧和红点蛤蚧药用价值。

蛤蚧(Gekko gecko)具有补肺滋胃，定喘止咳的功能,用于治疗虚喘气促，劳嗽咳血，阳痿遗精等症。长期以来，红点蛤蚧是否具有与黑点蛤蚧相似的药效存在着不同的看法。鉴于近年来广西黑点蛤蚧野生数量的骤减，产自东南亚地区的红点蛤蚧是目前蛤蚧药材市场上的主要流通品种，对红、黑点蛤蚧的药效，特别是红蛤蚧的药效进行科学评估显得十分必要。用SELDI蛋白图谱技术对蛤蚧进行真伪鉴定，该方法首先用30只黑点蛤蚧和30只红点蛤蚧的蛋白指纹图谱建立分类树模型，该模型经41例灰黑点斑蛤蚧和红点蛤蚧的盲筛以验证其可用性，结果表明，由1191 Da、1483 Da、6683 Da和25558 Da四个标志蛋白谱峰建立的模型对黑点蛤蚧诊断的准确性为100.000%，对红点蛤蚧诊断的准确性为95.455%，诊断的特异度和灵敏度的ROC值均为0.999，这为用蛋白质谱技术对动物中药进行规范化检测服务的可行性提供了参考。通过ELISA法测定生理盐水对照组、地塞米松干预组、黑点蛤蚧干预组和红点蛤蚧干预组（每组10只）的哮喘模型小鼠的肺泡灌洗液（bronchoalveolar lavage fluid，BALF）中IgE的变化、细胞染色计数法计算BALF中嗜酸性粒细胞（EOS）占白细胞比例变化以及哮喘症状变化情况，比较黑点蛤蚧与红点蛤蚧对哮喘的疗效，结果显示黑点蛤蚧对哮喘小鼠的药效高于红点蛤蚧。借鉴目前植物药材上“谱效”分析方法:灰色关联度分析、相关分析、回归分析等方法初步探索出一种适合动物药材蛤蚧“谱效”分析的方法，从而进一步评估了红、黑蛤蚧药用价值。