Effects of soluble proteins of Pheretima aspergillum prepared by different concoctions on HaCaT cells
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 as-sumed that the active ingredient mainly existed in the low molecular weight region.The main protein bands of low temperature ar-tillery 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 prod-uct 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.
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