不同炮制工艺制备广地龙可溶性蛋白对HaCaT细胞的影响  

Effects of soluble proteins of Pheretima aspergillum prepared by different concoctions on HaCaT cells

作　　者：宁盈悦 田林华 姚琳 冯丽娜 高辛 王伟明 董坤 NING Ying-yue;TIAN Lin-hua;YAO Lin;FENG Li-na;GAO Xin;WANG Wei-ming;DONG Kun(Heilongjiang Academy of Chinese Medicine Sciences,Harbin 150036,China)

机构地区：[1]黑龙江省中医药科学院,黑龙江哈尔滨150036

出　　处：《时珍国医国药》2024年第6期1402-1404,共3页Lishizhen Medicine and Materia Medica Research

基　　金：国家中药材产业技术体系专项项目(CARS-21);黑龙江省自然科学基金(LH2019H091)。

摘　　要：目的以鲜广地龙及其不同炮制品包括净制品、炒制品及低温炮制品为研究对象,进一步考察不同炮制工艺对广地龙中可溶性蛋白含量、种类及HaCaT细胞增殖的影响。方法采用Bradford法和SDS-PAGE法,检测广地龙不同炮制品中蛋白质的含量和种类;用CCK-8法进行活性筛选,测定其对HaCaT细胞增殖的影响结果4种广地龙炮制样品可溶性蛋白含量由高到低排序为:鲜品>低温炮制品>净制品>炒制品。鲜品的蛋白条带数目最多,主要集中在27.75~145.07KD低分子量区;低温炮制品与鲜品的蛋白质主带基本一致,净制品和炒制品低分子量区蛋白条带数目明显少于其他两种炮制品。不同广地龙样品给药48 h,净制品组HaCaT细胞基本死亡,炒制品组HaCaT细胞全部死亡,鲜品组和低温炮制组均能促进HaCaT细胞增殖,且鲜品组促细胞增殖效果更显著。结论广地龙中促进HaCaT细胞增殖的主要活性成分是存在于低分子量区的蛋白质,其促增殖活性与蛋白质的含量和种类呈正相关。Objective To investigate the effects of different processing techniques on the content,types of soluble protein and proliferation of HaCaT cells in fresh Pheretima aspergillum and its different prepared products,including purified,fried and low-temperature prepared products.Methods The content and types of proteins in different concoctions of Pheretima aspergillum were detected by Bradford's method and SDS-PAGE,and then screened for activity by CCK-8 method to determine their effects on the proliferation of HaCaT cells.Results The soluble protein contents of the four samples were analyzed by Bradford's method in the following order:fresh,low temperature processed,purified and fried.SDS-PAGE analysis showed that the fresh product had the highest number of protein bands,which were mainly concentrated in the range of 27.75 KD~145.07 KD,and it was assumed that the active ingredient mainly existed in the low molecular weight region.The main protein bands of low temperature artillery products and fresh products were basically the same,and the number of protein bands of net products and fried products was obviously less than that of the first two artillery processes.After 48h administration of different Pheretima aspergillum samples,HaCaT cells basically died in the purified product group,and all HaCaT cells died in the fried product group.Both the fresh product group and the low temperature processing group could promote HaCaT cell proliferation,and the fresh product group had a more significant effect on cell proliferation.Conclusion The main active components in Pheretima aspergillum that promote the proliferation of HaCaT cells are proteins present in the low molecular weight region,and their proliferative activities are positively correlated with the content and type of proteins.

关 键 词：广地龙 炮制方法 可溶性蛋白 HACAT细胞 

分 类 号：R283[医药卫生—中药学]