Andrographolide as An Anti-H1N1 Drug and the Mechanism Related to Retinoic Acid-Inducible Gene-I-Like Receptors Signaling Pathway

【摘要】 Objective:To observe the anti-virus effects of andrographolide(AD) on the retinoic acid-inducible gene-l(RIG-l)-like receptors(RLRs) signaling pathway when immunological cells were infected with H1N1.Methods:Leukomonocyte was obtained from umbilical cord blood by Ficoll density gradient centrifugation,and immunological cells were harvested after cytokines stimulation.Virus infected cell model was established by H1N1 co-cultured with normal human bronchial epithelial cell line(16HBE).The optimal concentration of AD was defined by methyl-thiazolyl-tetrazolium(MTT) assay.After the virus infected cell model was established,AD was added into the medium as a treatment intervention.After 24-h co-culture,cell supernatant was collected for interferon gamma(IFN-γ) and inteiieukin-4(IL-4) enzyme-linked immunosorbent assay(ELISA) detection while immunological cells for real-time polymerase chain reaction(RT-PCR).Results:The optimal concentration of AD for anti-virus effect was 250 μg/mL IL-4 and IFN-γ in the supernatant and mRNA levels in RLRs pathway increased when cells was infected by virus,RIG-I,IFN-β promoter stimulator-1(IPS-1),interferon regulatory factor(IRF)-γ,IRF-3 and nuclear transcription factor κB(NF-κB) mRNA levels increased significantly(P<0.05).When AD was added into co-culture medium,the levels of IL-4 and IFN-γ were lower than those in the non-interference groups and the mRNA expression levels decreased,RIG-I,IPS-1,IRF-γ,IRF-3 and NFk B decreased significantly in each group with significant statistic differences(P<0.05).Conclusions:The RLRs mediated viral recognition provided a potential molecular target for acute viral infections and andrographolide could ameliorate H1N1 virus-induced cell mortality.And the antiviral effects might be related to its inhibition of viral-induced activation of the RLRs signaling pathway.更多还原

【Abstract】 Objective:To observe the anti-virus effects of andrographolide(AD) on the retinoic acid-inducible gene-l(RIG-l)-like receptors(RLRs) signaling pathway when immunological cells were infected with H1N1.Methods:Leukomonocyte was obtained from umbilical cord blood by Ficoll density gradient centrifugation,and immunological cells were harvested after cytokines stimulation.Virus infected cell model was established by H1N1 co-cultured with normal human bronchial epithelial cell line(16HBE).The optimal concentration of AD was defined by methyl-thiazolyl-tetrazolium(MTT) assay.After the virus infected cell model was established,AD was added into the medium as a treatment intervention.After 24-h co-culture,cell supernatant was collected for interferon gamma(IFN-γ) and inteiieukin-4(IL-4) enzyme-linked immunosorbent assay(ELISA) detection while immunological cells for real-time polymerase chain reaction(RT-PCR).Results:The optimal concentration of AD for anti-virus effect was 250 μg/mL IL-4 and IFN-γ in the supernatant and mRNA levels in RLRs pathway increased when cells was infected by virus,RIG-I,IFN-β promoter stimulator-1(IPS-1),interferon regulatory factor(IRF)-γ,IRF-3 and nuclear transcription factor κB(NF-κB) mRNA levels increased significantly(P<0.05).When AD was added into co-culture medium,the levels of IL-4 and IFN-γ were lower than those in the non-interference groups and the mRNA expression levels decreased,RIG-I,IPS-1,IRF-γ,IRF-3 and NFk B decreased significantly in each group with significant statistic differences(P<0.05).Conclusions:The RLRs mediated viral recognition provided a potential molecular target for acute viral infections and andrographolide could ameliorate H1N1 virus-induced cell mortality.And the antiviral effects might be related to its inhibition of viral-induced activation of the RLRs signaling pathway.更多还原