山柰酚通过下调NF-κB信号通路减轻猪源甲型H9N2流感病毒所致小鼠急性肺损伤
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摘要
目的:探讨山柰酚是否通过下调转录因子NF-κB信号通路的表达而保护感染猪源甲型H9N2流感病毒引起急性肺损伤的小鼠。方法:猪源甲型H9N2流感病毒感染BALB/c小鼠建立急性肺损伤模型,山柰酚干预后检测肺湿重与干重比,观察肺组织的病理学变化,检测支气管肺泡灌洗液内炎性细胞数量以及肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)和白细胞介素1β(IL-1β)含量同时检测肺组织匀浆中超氧化物歧化酶(SOD)和髓过氧化物酶(MPO)活性以及丙二醛(MDA)含量,Western blot检测小鼠肺内NF-κB P65的表达,ELISA检测小鼠肺组织匀浆细胞核提取物中NF-κB P65和NF-κB P50的核转位。结果:山柰酚能降低小鼠死亡率,改善肺组织的病理学变化和肺水肿程度,并能降低肺内巨噬细胞、淋巴细胞和中性粒细胞等炎性细胞的数量同时降低TNF-α、IL-6、IL-1β和MDA的含量,抑制MPO的活性并升高SOD的活性。另外,山柰酚可以下调NF-κB P65的表达增加细胞核提取物中NF-κB P65和NF-κB P50的核转位。结论:山柰酚通过下调NF-κB信号通路的表达从而降低猪源甲型H9N2流感病毒所致急性肺损伤小鼠的炎症程度和氧化应激损伤,最终减轻流感病毒所致的急性肺损伤。
AIM:To investigate whether kaempferol protects against acute lung injury induced by swine-origin influenza A H9N2 virus via down-regulation of NF-κB signaling pathway.METHODS:BALB/c mice were used to establish the animal model of acute lung injury by nasal inoculation of swine-origin influenza A H9N2 virus.After the intervention with kaempferol,the pulmonary edema was evaluated by determining the lung wet weight/dry weight(W/D) ratio,the pathological changes of the lung tissues were observed,the concentrations of TNF-α,IL-1β and IL-6 in the bronchoalveolar lavage fluid(BALF) were measured,and superoxide dismutase(SOD) activity,myeloperoxidase(MPO) activity and MDA content in the homogenate of the lung tissues were detected.NF-κB P65 levels were determined by Western blot,and the NF-κB P65 and NF-κB P50 nuclear translocation in the nuclear extracts from mouse lung tissue homogenate was detected by ELISA.RESULTS:Treatment with kaempferol decreased the morality of infected mice,and significantly prolonged the survival time of the infected mice.Kaempferol also relieved the pathological changes of the lung tissues,the lung W/D ratio and the lung index in swine-origin influenza A H9N2 virus-infected mice.Treatment with kaempferol significantly decreased the infiltration of inflammatory cells including macrophages,lymphocytes and neutrophils in the BALF.The levels of TNF-α,IL-6,IL-1β and MDA and the activity of MPO were also decreased.Treatment with kaempferol also significantly increased the SOD activity.NF-κB P65 levels were decreased,and the NF-κB P65 and NF-κB P50 nuclear translocation in the nuclear extracts from the mouse lung tissue homogenate were also decreased by treatment with kaempferol.CONCLUSION:The protective effect of kaempferol on the mice with acute lung injury induced by swine-origin influenza A H9N2 virus is related to suppression of the oxidative stress and inflammatory responses by down-regulation of NF-κB signaling pathway.
AIM:To investigate whether kaempferol protects against acute lung injury induced by swine-origin influenza A H9N2 virus via down-regulation of NF-κB signaling pathway.METHODS:BALB/c mice were used to establish the animal model of acute lung injury by nasal inoculation of swine-origin influenza A H9N2 virus.After the intervention with kaempferol,the pulmonary edema was evaluated by determining the lung wet weight/dry weight(W/D) ratio,the pathological changes of the lung tissues were observed,the concentrations of TNF-α,IL-1β and IL-6 in the bronchoalveolar lavage fluid(BALF) were measured,and superoxide dismutase(SOD) activity,myeloperoxidase(MPO) activity and MDA content in the homogenate of the lung tissues were detected.NF-κB P65 levels were determined by Western blot,and the NF-κB P65 and NF-κB P50 nuclear translocation in the nuclear extracts from mouse lung tissue homogenate was detected by ELISA.RESULTS:Treatment with kaempferol decreased the morality of infected mice,and significantly prolonged the survival time of the infected mice.Kaempferol also relieved the pathological changes of the lung tissues,the lung W/D ratio and the lung index in swine-origin influenza A H9N2 virus-infected mice.Treatment with kaempferol significantly decreased the infiltration of inflammatory cells including macrophages,lymphocytes and neutrophils in the BALF.The levels of TNF-α,IL-6,IL-1β and MDA and the activity of MPO were also decreased.Treatment with kaempferol also significantly increased the SOD activity.NF-κB P65 levels were decreased,and the NF-κB P65 and NF-κB P50 nuclear translocation in the nuclear extracts from the mouse lung tissue homogenate were also decreased by treatment with kaempferol.CONCLUSION:The protective effect of kaempferol on the mice with acute lung injury induced by swine-origin influenza A H9N2 virus is related to suppression of the oxidative stress and inflammatory responses by down-regulation of NF-κB signaling pathway.
引文
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[11]Zhang A,Wang S,Zhang J,et al.Genipin alleviates LPS-induced acute lung injury by inhibiting NF-κB and NLRP3 signaling pathways[J].Int Immunopharmacol,2016,38:115-119.
[12]Wu H,Zhao G,Jiang K,et al.Plantamajoside ameliorates lipopolysaccharide-induced acute lung injury via suppressing NF-κB and MAPK activation[J].Int Immunopharmacol,2016,35:315-322.
[13]Kasahara DI,Poynter ME,Othman Z,et al.Acrolein inhalation suppresses lipopolysaccharide-induced inflammatory cytokine production but does not affect acute airways neutrophilia[J].J Immunol,2008,181(1):736-745.
[14]Chen X,Yang X,Liu T,et al.Kaempferol regulates MAPKs and NF-κB signaling pathways to attenuate LPSinduced acute lung injury in mice[J].Int Immunopharmacol,2012,14(2):209-216.
[15]Garcia-Mediavilla V,Crespo I,Collado PS,et al.The anti inflammatory flavones quercetin and kaempferol cause inhibition of inducible nitric oxide synthase,cyclooxygenase-2 and reactive C-protein,and down-regulation of the nuclear actor kappaB pathway in Chang Liver cells[J].Eur J Pharmacol,2007,557(2-3):221-229.
[16]何煜舟,丁美萍,汪云开,等.山萘酚对缺血再灌注脑损伤大鼠的保护作用[J].中华中医药学刊,2009,27(8):1673-1675.
[17]Kim JM,Lee EK,Kim DH,et al.Kaempferol modulates pro-inflammatory NF-κB activation by suppressing advanced glycation endproducts-induced NADPH oxidase[J].Age(Dordr),2010,32(2):197-208.
[18]Ehrhardt C,R(u|¨)ckle A,Hrincius ER,et al.The NF-κB inhibitor SC75741 efficiently blocks influenza vims propagation and confers a high barrier for development of viral resistance[J].Cell Microbiol,2013,15(7):1198-1211.
[19]Mazur I,Wurzer WJ,Ehrhardt C,et al.Acetylsalicylic acid(ASA)blocks influenza vims propagation via its NF-κB inhibiting activity[J].Cell Microbiol,2007,9(7):1683-1694.
[2]Wei D,Huang ZH,Zhang RH,et al.Roles of p38MAPK in the regulation of the inflammatory response to swine influenza virus-induced acute lung injury in mice[J].Acta Virol,2014,58(4):374-379.
[3]Zhang RH,Cui HY,Xu MJ,et al.Molecular characterization and pathogenicity of swine influenza H9N2 subtype vims A/swine/HeBei/012/2008/(H9N2)[J].Acta Virol,2011,55(3):219-226.
[4]Salomon R,Hoffmann E,Webster RG.Inhibition of the cytokine response does not protect against lethal H5N1 influenza infection[J].Proc Natl Acad Sci U S A,2007,104(30):12479-12481.
[5]王国华,王存连,田树飞,等.肌肽对H9N2亚型猪流感病毒诱导的小鼠急性肺损伤的抗氧化效果[J].中国病理生理杂志,2013,29(12):2235-2239.
[6]张瑞华,王存连,徐彤,等.N-乙酰半胱氨酸对H9N2猪流感病毒所致急性肺损伤小鼠的保护作用[J].中国病理生理杂志,2014,30(4):698-705.
[7]Nirmala P,Ramanathan M.Effect of kaempferol on lipid peroxidation and antioxidant status in 1,2-dimethyl hydrazine induced colorectalcarcinoma in rats[J].Eur J Pharmacol,2011,654(1):75-79.
[8]Hamalainen M,Nieminen R,Vuorela P,et al.Anti-inflammatory effects of flavonoids:genistein,kaempferol,quercetin,and daidzein inhibit STAT-1 and NF-κB activations,whereas flavone,isorhamnetin,naringenin,and pelargonidin inhibit only NF-κB activation along with their inhibitory effect on iNOS expression and NO production in activated macrophages[J].Mediators Inflamm,2007,2007:45673.
[9]Tsai FJ,Lin CW,Lai CC,et al.Kaempferol inhibits enterovirus 71 replication and internal ribosome entry site(IRES)activity through FUBP and HNRP proteins[J].Food Chem,2011,128(2):312-322.
[10]Yu X,Yu S,Chen L,et al.Tetrahydroberberrubine attenuates lipopolysaccharide-induced acute lung injury by down-regulating MAPK,AKT,and NF-κB signaling pathways[J].Biomed Pharmacother,2016,82:489-497.
[11]Zhang A,Wang S,Zhang J,et al.Genipin alleviates LPS-induced acute lung injury by inhibiting NF-κB and NLRP3 signaling pathways[J].Int Immunopharmacol,2016,38:115-119.
[12]Wu H,Zhao G,Jiang K,et al.Plantamajoside ameliorates lipopolysaccharide-induced acute lung injury via suppressing NF-κB and MAPK activation[J].Int Immunopharmacol,2016,35:315-322.
[13]Kasahara DI,Poynter ME,Othman Z,et al.Acrolein inhalation suppresses lipopolysaccharide-induced inflammatory cytokine production but does not affect acute airways neutrophilia[J].J Immunol,2008,181(1):736-745.
[14]Chen X,Yang X,Liu T,et al.Kaempferol regulates MAPKs and NF-κB signaling pathways to attenuate LPSinduced acute lung injury in mice[J].Int Immunopharmacol,2012,14(2):209-216.
[15]Garcia-Mediavilla V,Crespo I,Collado PS,et al.The anti inflammatory flavones quercetin and kaempferol cause inhibition of inducible nitric oxide synthase,cyclooxygenase-2 and reactive C-protein,and down-regulation of the nuclear actor kappaB pathway in Chang Liver cells[J].Eur J Pharmacol,2007,557(2-3):221-229.
[16]何煜舟,丁美萍,汪云开,等.山萘酚对缺血再灌注脑损伤大鼠的保护作用[J].中华中医药学刊,2009,27(8):1673-1675.
[17]Kim JM,Lee EK,Kim DH,et al.Kaempferol modulates pro-inflammatory NF-κB activation by suppressing advanced glycation endproducts-induced NADPH oxidase[J].Age(Dordr),2010,32(2):197-208.
[18]Ehrhardt C,R(u|¨)ckle A,Hrincius ER,et al.The NF-κB inhibitor SC75741 efficiently blocks influenza vims propagation and confers a high barrier for development of viral resistance[J].Cell Microbiol,2013,15(7):1198-1211.
[19]Mazur I,Wurzer WJ,Ehrhardt C,et al.Acetylsalicylic acid(ASA)blocks influenza vims propagation via its NF-κB inhibiting activity[J].Cell Microbiol,2007,9(7):1683-1694.