Anti-influenza Effects Of Rosemary Acid Glycosides Extacted From Sarcandra Glabra In Vitro

ObjectiveTo evaluate the anti-influenza effects of rosemary acid glycosides extacted from Sarcandra glabra before,researches were performed in vitro.Methods1.Cytotoxic effects of rosemary acid glycosides(RAG)extracted from Sarcandra glabra on MDCK and A549 cells were determined using CCK8(Cell Counting Kit-8)method.2.Oseltamivir was used as positive reference drug,anti-H1N1 activity of RAG were examined by NA assay(NA-FIuorTM Influenza Neuraminidase Assay Kit)on A549 cells in 4 different treat models.3.Oseltamivir was used as positive reference drug,IL-6 and IP-10 ELISA kit were used to measure the levels of inflammatory cytokine in A549 cells treated with RAG after viral infection.4.Oseltamivir was used as positive reference drug,anti-H1N1 and H9N2 activity of RAG on A549 cells were examined by using CCK8 and NA assay.5.Oseltamivir was used as positive reference drug,anti-H1N1 activity of RAG on MDCK and A549 cells were detected by CCK8 and NA assay.6.Ribavirin was used as positive reference drug,anti-H1N1 activity of RAG on embryonated eggs was evaluated by observing survival rate and hemagglutination test.7.To understand the mechanisms involved in the antiviral effect,anti-H1N1 activity of RAG were examined by NA assay kit on A549 cells in time-of-addition(-2h～-1h,-1h～0h,0h～2h,2h～4h,4h～6h)assay.Results1.RAG extracted from Sarcandra glabra shows no cytotoxicity on MDCK and A549 cells,and the 50%toxic concentration(TC50)values are higher than 250μM.2.RAG showed effective anti-H1N1 activity on A549 cells in treatment model,with 50%inhibiting concentration(IC50)value of 102.2μM.It showed some inhibitory effect in premix model and no anti-H1N1 activity in prevention model.3.RAG inhibited the expression of IL-6 and IP-10 in A549 cells infected by H1N1 influenza virus.4.RAG showed some anti-H9N2 activity on A549 cells in treatment model.5.RAG showed effective anti-HIN1 activity on A549 cells and MDCK cells in treatment model,with 50%inhibiting concentration(IC50)value of 102.2μM and 85.37μM respectively.6.RAG showed potential anti-HIN1 activity on embryonated eggs.7.RAG did not exert antiviral activity when it was added before infection.A significant inhibition of virus replication was observed when RAG was added to the cells within the time frame of Oh to 2h.ConclusionIn 0.39μM～200μM concentration,anti-influenza effect of RAG can be obsearved in different administration modes,and influenza virus replication activity is ininhibited most when RAG is added after infection.Under this mode,RAG shows effective anti-H9N2 activity on A549 cells and effective anti-H1N1 activity on MDCK cells.Potential anti-H1N1 activity is also obsearved on embryonated eggs.The result of time-of-addition assay shows that RAG is effective during early stage like viral endocytosis,and further study should be taken to clear its targets.