Study On The Bioactivity And Preparation Of Periplaneta Americana Extract And Lysophosphatidylcholine Derivatives

Objective: To study the preparation process of Periplaneta americana extract,its antifungal activity and activation of NKT cells,and to study the immune activity of lysophosphatidylcholine derivatives on NKT cells.At the same time,compound Periplaneta americana tablets were prepared with Periplaneta americana as the main drug,and its quality standard was preliminarily studied.Methods: Different extraction methods and solvents were used to extract Periplaneta americana.The antifungal activity of Periplaneta americana extract was studied by microbiological assay.Enzyme linked immunosorbent assay(ELISA)was used to study the activation of NKT cells by different extracts of Periplaneta americana.The activation of NKT by lysophosphatidylcholine(LPC)derivatives was studied by enzyme-linked immunosorbent assay.Single factor and orthogonal experiments were used to optimize the molding process of compound Periplaneta americana tablets.The appearance,weight difference and disintegration time limit were examined according to the general principles of preparation in Chinese Pharmacopoeia;the TLC identification methods of Salvia miltiorrhiza,Hawthorn,Panax notoginseng and Periplaneta americana were established;the contents of TanshinoneⅡA and Salvianolic acid B were determined by HPLC,and the methodology was investigated.Results: Five different extracts of Periplaneta americana were obtained.Through antifungal activity study,it was found that Chloroform Methanol(1:1)extract,n-butanol extract,n-butanol extract with fluconazole had combined antibacterial effect on Candida albicans resistant strains.Enzyme linked immunosorbent assay(ELISA)showed that petroleum ether extract,Chloroform Methanol(1:1)extract and n-butanol extract of Periplaneta americana could activate NKT cells at a certain concentration.Lysophosphatidylcholine(LPC)derivatives can activate NKT cells.Compound Periplaneta americana tablets were prepared with Periplaneta americana as the main drug.The preparation process was as follows: Periplaneta americana and Salvia miltiorrhiza were taken,added 10 times of 70% ethanol,refluxed twice,1 hour each time,filtered;the filtrate was recovered with ethanol for standby;the filter residue was mixed with hawthorn,added 10 times of water,extracted twice,1.5 hours each time,filtered.After ethanol recovery,the filtrate and water extract were mixed and concentrated to a clear paste with a relative density of 1.25(80 ℃),mixed with Panax notoginseng fine powder,Pueraria lobata fine powder and 4% magnesium oxide(based on Periplaneta americana),dried and crushed into fine powder,and then 3% povidone K30 was added to make granules,3% talcum powder was added to mix well,and the tablets were obtained.According to the quality standard study,the disintegration time limit and weight difference meet the requirements of Pharmacopoeia;TLC identification meet the requirements;the content of Tanshinone ⅡA and Salvianolic Acid B in compound Periplaneta Americana tablets should not be less than 0.12 mg and 1.4mg respectively.Conclusion:The extracts of Periplaneta americana have antibacterial and immunomodulatory effects.Some of them can activate NKT cells and can be used as natural products of NKT cells.Lysophosphatidylcholine(LPC)derivatives can also activate NKT cells,and can be used as the chemosynthetic exogenous antigens of NKT cells.The study of NKT activity of these two kinds of substances will greatly promote the study of exogenous antigen of NKT cells.The results showed that the preparation process of compound Periplaneta americana tablets was stable and feasible,and the quality standard TLC identification and content determination method had good reproducibility,which could be used for the quality control of compound Periplaneta americana tablets.