Isolation And Anti-inflammation Activity Of Diterpenoid Alkaloids From Tibetan Herbal Medicine Ponka

Ponka is a kind of commonly used Tibetan medicine,referring to the dried whole plant of Aconitum tanguticum(Maxim.)Stapf or Aconitum navicuare(Bruhl.)Stapf.It has the effects of clearing away heat and detoxifying,and is often used for treating fever caused by infectious diseases,flu and food poisoning,etc.Modern studies showed that Ponka has anti-inflammatory,antibacterial,antiviral activities.It mainly contains diterpenoid alkaloids,flavonoids and phenolic acids.Firstly,this paper explore the active components and mechanism of antiinfluenza virus,anti-inflammation and anti-Alzheimer’s Disease activities of Ponka by network pharmacological analysis.As a result,total of 73 active components,20 core ingredients,841 action targets,94 intersection targets of Ponka and influenza virus,142 intersection targets of Ponka and anti-inflammation,227 intersection targets of Ponka and anti-Alzheimer’s Disease were retrieved.GO functional analysis revealed 181 biological processes,42 molecular functions and 25 cell components of Ponka against influenza virus.KEGG enrichment analysis obtained 84 anti-influenza virus signaling pathways,mainly involving TNF,Hepatitis C and cancer signaling pathways.Go functional analysis revealed 232 biological processes,52 molecular functions and 27 cell components of Ponka against inflammation.KEGG enrichment analysis showed 86 anti-inflammatory signaling pathways,mainly involving NFKappa B,TNF and HIF-1 signaling pathways.Go functional analysis revealed 333 biological processes,82 molecular functions and 59 cell components of Ponka against Alzheimer’s Disease.KEGG enrichment analysis showed 105 anti-Alzheimer’s Disease signaling pathways,mainly involving Alzheimer’s Disease and ErbB signaling pathways.The Potential active components are diterpenoid alkaloids and phenolic acids.Therefore,on the basis of the previous research,this paper continued to explore the diterpenoid alkaloids of Ponka.By using a variety of modern chromatographic and spectral techniques and comparison with literature values,total of 7 compounds were isolated and identified,and they were Tanguticinine F(1),Tanguticinine G(2),5-deoxyanthoroidine B(3),N-oxide 5-deoxyanthoroidine B(4),Navirine D(5),anthoroidine B(6)and 6-acetyl heteratisine(7).Among them,compounds 1～5 were new compounds.Using the RAW264.7 cells induced by lipopolysaocharides(LPS)as the in vitro anti-inflammatory activity screening model,the inhibitory effects of the seven diterpene alkaloids and the other fifteen diterpenoid alkaloids prepared before,on the release of nitric oxide(no)and TNF-α(tumor necrosis factor)were studied.The results showed that compound 1,3,5,6 had significant inhibition effect on the NO production by LPS-induced macrophage RAW264.7 cell,with EC50 of 344.579,676.408,257.214 and 357.680μM respectively.But none of the seven compounds showed significant inhibition on the TNF-αsecretion.The results of this study not only elucidate the basis of Ponka against heat and toxic materials,but also enrich the structure types of natural products and provide a basis for the research and development of new drugs.