清透邪热方对甲型H1N1流感病毒感染Ana-1细胞TLR-7、MyD88及NF-κB mRNA及蛋白表达的影响

冯丽谦; 杨丽丽; 李明真; 孙雨; 刘涛

清透邪热方对甲型H1N1流感病毒感染Ana-1细胞TLR-7、MyD88及NF-κB mRNA及蛋白表达的影响

1.
南京中医药大学基础医学院，江苏南京 210023
2.
南京中医药大学第一临床医学院，江苏南京 210023
3.
南京中医药大学护理学院，江苏南京 210023

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出版历程
- 刊出日期: 2017-05-10

Effects of Qingtouxiere Decoction on Expression of TLR-7， MyD88 and NF-κB mRNA and Protein in Ana-1 Cells of Influenza A H1N1 Influenza Virus

1.
The Basic Medical College, Nanjing University of Chinese Medicine, Nanjing, 210023, China
2.
The First Clinical Medical School, Nanjing University of Chinese Medicine, Nanjing, 210023, China
3.
School of Nursing, Nanjing University of Chinese Medicine, Nanjing, 210023, China

摘要

摘要: 目的研究清透邪热方体外抗甲型H1N1流感病毒效应机制。方法将清透邪热方制备成0.388、0.194、0.097、0.0485、0.02425mg/mL不同浓度，用H1N1甲型流感病毒感染小鼠Ana-1巨噬细胞，给予不同浓度清透邪热方及达菲进行干预，检测肿瘤坏死因子α（TNF-α）、干扰素诱导蛋白-10（IP-10）和白细胞介素-6（IL-6）表达，Toll样受体7（TLR7）及其下游髓样分化因子88(MyD88)、核因子-κB（NF-κB）mRNA和蛋白表达。结果清透邪热方能够抑制甲型H1N1流感病毒感染Ana-1细胞TNF-α、IP-10、IL-6的表达；其中清透邪热方0.02425mg/mL组降低H1N1病毒感染组TNF-α、IL-6显著（P＜0.01）；清透邪热方0.097mg/mL组降低IP-10显著（P＜0.01）。清透邪热方能够抑制H1N1流感病毒感染后Ana-1细胞后TLR-7、MyD88及NF-κB mRNA及蛋白表达升高；其中清透邪热方0.0485mg/mL组抑制H1N1病毒感染组TLR-7 mRNA、MyD88 mRNA表达显著（P＜0.01）；清透邪热方0.02425mg/mL组抑制H1N1病毒感染组NF-κB mRNA表达显著（P＜0.01）；清透邪热方0.0485mg/mL组抑制H1N1病毒感染组TLR-7蛋白表达升高显著（P＜0.01），清透邪热方0.388mg/mL组抑制H1N1病毒感染组MyD88蛋白、NF-κB蛋白表达显著（P＜0.01）。结论清透邪热方通过抑制甲型H1N1流感病毒感染Ana-1细胞TLR-7、MyD88及NF-κB mRNA及蛋白表达升高，减少TNF-α、IP-10和IL-6表达，起到抗病毒作用。
- 清透邪热方 /
- 甲型H1N1流感病毒 /
- 小鼠Ana-1巨噬细胞 /
- TLR-7/MyD88/NF-κB通路
Abstract: OBJECTIVE To observe the effect mechanism of Qingtouxiere Decoction on anti-influenza A H1N1 influenza virus in vitro. METHODS The macrophages were stimulated with FM1 in the lungs of influenza A virus， respectively， with different concentrations of 0.388mg/mL， 0.194mg/mL， 0.097mg/mL， 0.0485mg/mL and 0.02425mg/mL. The expression of TNF-α， IP-10 and IL-6， TLR-7， mRNA and protein expression of MyD88 and NF-κB. RESULTS The expression of TNF-α， IP-10 and IL-6 in macrophages of Ana-1 mice was inhibited by Qingtouxiere Decoction， and the infection of H1N1 virus was decreased in 0.02425mg/mL group (P<0.01). The level of TNF-α was significantly lower in the group of Qingtouxiere Decoction 0.097mg/mL (P <0.01). The expression of TLR-7， MyD88 and NF-κB mRNA and protein were increased after Ana-1 macrophage infection in H1N1 influenza virus group. The expression of TLR-7， MyD88 and NF-κB mRNA was increased by 0.0485mg/mL group (P<0.01). The expression of MyD88 mRNA in the H1N1 infected group was significantly higher than that in the control group (P <0.01). The expression of TLR-7 mRNA was significantly inhibited The expression of NF-κB mRNA in H1N1 virus group was significantly higher than that in H1N1 virus group (P<0.01). The expression of TLR-7 protein in H1N1 virus group was significantly higher than that in H1N1 virus group(P<0.01). The expression of NF-κB in the H1N1 infected group was significantly higher than that in the H1N1 virus group (P<0.01). The expression of NF-κB in the H1N1 infected group was significantly higher than that in the control group (P<0.01). CONCLUSION The activation of TLR-7/MyD88/NF-κB signaling pathway induced by H1N1 influenza virus can inhibit the expression of downstream inflammatory factors and play the antiviral effect.
- Qingtouxiere Decoction /
- H1N1 influenza virus /
- mouse Ana-1 cells /
- TLR-7/MyD88/NF- B pathway