三白草水提液抗肠道病毒71型作用初探
|
摘要
目的:初步探讨三白草水提液抗肠道病毒71型(EV71)的活性及其机制。方法:采用EV71感染非洲绿猴肾上皮(Vero细胞)模型,以不同浓度的三白草水提液提前预处理细胞,结合MTT方法检测细胞活力,采用TCID50的方法检测子代病毒的释放,评价三白草水提物抗EV71病毒能力。应用Realtime PCR的方法检查三白草水提物对炎症因子产生的作用。以免疫印迹法检测三白草水提物对NF-κB信号通路的影响。利巴韦林为阳性对照。结果:三白草水提物0. 1、0. 03、0. 015 mg/ml的浓度下能够显著抑制EV71感染诱导的细胞死亡,并且这种抑制作用具有剂量依赖性。与病毒组相比,三白草水提液在0. 1、0. 03以及0. 015 mg/ml的浓度下分别将病毒滴度降低了1. 60 logs、1. 85 logs和3. 45 logs。利巴韦林将病毒滴度降低了3. 54 logs。Realtime PCR的结果显示,三白草水提液能够明显降低EV71诱导的IL-1β、IL-6以及IL-8 m RNA的表达。提取核蛋白,免疫印迹检测结果显示,三白草水提物能够阻断EV71诱导的NF-κB核转移。结论:三白草水提液在体外具有抗EV71病毒复制的活性,并且可能通过抑制NF-κB核转移抑制炎症因子的产生。
Objective: To evaluate the antiviral activity of water extracts of Saururus chinensis( Lour.) Baill on enterovirus 71 infection. Methods: Vero cells were infected with EV71. Then the cells were treated with or without water extracts of Saururus chinensis( Lour.) Baill. Ribavirin was served as a positive reference drug. Cell viability was monitored by MTT assay and virus titers were tested by TCID50 method. The expression of cytokines was detected by Realtime PCR,while nuclear translocation of p-65/NF-κB were examined by Western blot assay. Results: Water extracts of Saururus chinensis( Lour.) Baill significantly inhibited EV71 induced cell death in a dose dependent way. Compared with the virus group,water extract at 0. 1 mg/ml,0. 03 mg/ml and 0. 015 mg/ml decreased the virus titers by 1. 60 logs,1. 85 logs and 3. 45 logs,respectively,while Ribavirin downregulated the virus titer by 3. 54 logs. Moreover,water extract of Saururus chinensis( Lour.) Baill inhibited the m RNA level of IL-1β,IL-6 and IL-8. The results also showed that Water extracts of Saururus chinensis( Lour.) Baill can notably block the neuclear translocation of p-65/NF-κB,suggesting that EV71 blocked the expression of cytokines by bloking NF-κB signal pathway. Conclusion: Water extract of Saururus chinensis( Lour.) Baill possessed anti-EV71 activity and inhibited the expressions of cytokines by blocking the nuclear translocation of p-65/NF-κB.
Objective: To evaluate the antiviral activity of water extracts of Saururus chinensis( Lour.) Baill on enterovirus 71 infection. Methods: Vero cells were infected with EV71. Then the cells were treated with or without water extracts of Saururus chinensis( Lour.) Baill. Ribavirin was served as a positive reference drug. Cell viability was monitored by MTT assay and virus titers were tested by TCID50 method. The expression of cytokines was detected by Realtime PCR,while nuclear translocation of p-65/NF-κB were examined by Western blot assay. Results: Water extracts of Saururus chinensis( Lour.) Baill significantly inhibited EV71 induced cell death in a dose dependent way. Compared with the virus group,water extract at 0. 1 mg/ml,0. 03 mg/ml and 0. 015 mg/ml decreased the virus titers by 1. 60 logs,1. 85 logs and 3. 45 logs,respectively,while Ribavirin downregulated the virus titer by 3. 54 logs. Moreover,water extract of Saururus chinensis( Lour.) Baill inhibited the m RNA level of IL-1β,IL-6 and IL-8. The results also showed that Water extracts of Saururus chinensis( Lour.) Baill can notably block the neuclear translocation of p-65/NF-κB,suggesting that EV71 blocked the expression of cytokines by bloking NF-κB signal pathway. Conclusion: Water extract of Saururus chinensis( Lour.) Baill possessed anti-EV71 activity and inhibited the expressions of cytokines by blocking the nuclear translocation of p-65/NF-κB.
引文
[1]谢金炎,程安春,汪铭书.肠道病毒与宿主固有免疫系统相互作用的研究进展[J].中国免疫学杂志,2017,33(8):1263-1266.Xie JY,Cheng AC,Wang MS. Research progress on the interaction of Enterovirus and host innate immune system[J]. Chin J Immunol,2017,33(8):1263-1266.
[2] Chung BS,Shin MG. Dictionary of Korean Folk Medicine[M].Young Lim Sa:Seoul,Korea,1990.
[3] Zhang ZL,Zuo YM,Lu XU,et al.Studies on chemical components of flavonoids in aerial part of Saururus chinensis[J].Chin Tradition Herbal Drugs,2011,42(8):1490-1493.
[4] Bang YH,Lee JH,Nam JB,et al.Lignans from Saururus chinensis,inhibiting the transcription factor NF-κB[J]. Phytochemistry,2003,64(3):765-771.
[5] Khan MT,Ather A,Thompson KD,et al. Extracts and molecules from medicinal plants against herpes simplex viruses[J]. Antivir Res,2005,67(2):107-119.
[6] Su YL,Rhim JY,Park WB. Antiherpetic activities of flavonoids against herpes simplex virus type 1(HSV-1)and type 2(HSV-2)in vitro[J].Archives Pharmacal Res,2005,28(11):1293-301.
[7] Mosmann T. Rapid colorimetric assay for cellular growth and survival:application to proliferation and cytotoxicity assays[J]. J Immunol Methods,1983,65(1/2):55.
[8] Chen X,Wang C,Xu L,et al.A laboratory evaluation of medicinal herbs used in china for the treatment of hand,foot,and mouth disease[J]. Evid Based Complement Alternat Med, 2013,2013:504563.
[9] Lin TY,Hsia SH,Huang YC,et al.Proinflammatory cytokine reactions in enterovirus 71 infections of the central nervous system.[J].Clin Infect Dis,2003,36(3):269-274.
[10] Xun G,Zhou J,Zhu W,et al.Excessive proinflammatory cytokine and chemokine responses of human monocyte-derived macrophages to enterovirus 71 infection[J]. BMC Infect Dis,2012,12(1):224-224.
[11] Chen CS,Yao YC,Lin SC,et al. Retrograde axonal transport:a major transmissionroute of ente rovirus 71 in mice[J]. J Virol,2007,81(17):8996-9003.
[12] Lin TY,Chang LY,Hsia SH,et al.The 1998 enterovirus 71 outbreak in Taiwan:pathogenesis and management[J]. Clin Infect Dis,2002,34(Suppl 2):S52-S57.
[13]赵瑞玲,毛秀娣,张丽锋.利巴韦林在儿科临床应用的安全性评价[J].国际药学研究杂志,2016,43(4):615-620.Zhao RL,Mao XT,Zhang LF. Safety evaluation of ribavirin in clinical practice in paediatrics[J]. J Int Pharmaceut Res,2016,43(4):615-620.
[14]田蕾,李晓艳,徐云霞,等.三白草抗单纯疱疹病毒作用及机制[J].中国中药杂志,2012,37(11):1642-1645.Tian L,Li XY,Xu YX,et al. Eeffect and mechanism of Saururus chinensis against herpes dimplex virus[J].China J Chin Material Medica,2012,37(11):1642-1645.
[15] Khong WX,Foo DG,Trasti SL,et al.Sustained high levels of interleukin-6 contribute to the pathogenesis of enterovirus 71 in a neonate mouse model[J].J Virol,2011,85(7):3067-3076.
[2] Chung BS,Shin MG. Dictionary of Korean Folk Medicine[M].Young Lim Sa:Seoul,Korea,1990.
[3] Zhang ZL,Zuo YM,Lu XU,et al.Studies on chemical components of flavonoids in aerial part of Saururus chinensis[J].Chin Tradition Herbal Drugs,2011,42(8):1490-1493.
[4] Bang YH,Lee JH,Nam JB,et al.Lignans from Saururus chinensis,inhibiting the transcription factor NF-κB[J]. Phytochemistry,2003,64(3):765-771.
[5] Khan MT,Ather A,Thompson KD,et al. Extracts and molecules from medicinal plants against herpes simplex viruses[J]. Antivir Res,2005,67(2):107-119.
[6] Su YL,Rhim JY,Park WB. Antiherpetic activities of flavonoids against herpes simplex virus type 1(HSV-1)and type 2(HSV-2)in vitro[J].Archives Pharmacal Res,2005,28(11):1293-301.
[7] Mosmann T. Rapid colorimetric assay for cellular growth and survival:application to proliferation and cytotoxicity assays[J]. J Immunol Methods,1983,65(1/2):55.
[8] Chen X,Wang C,Xu L,et al.A laboratory evaluation of medicinal herbs used in china for the treatment of hand,foot,and mouth disease[J]. Evid Based Complement Alternat Med, 2013,2013:504563.
[9] Lin TY,Hsia SH,Huang YC,et al.Proinflammatory cytokine reactions in enterovirus 71 infections of the central nervous system.[J].Clin Infect Dis,2003,36(3):269-274.
[10] Xun G,Zhou J,Zhu W,et al.Excessive proinflammatory cytokine and chemokine responses of human monocyte-derived macrophages to enterovirus 71 infection[J]. BMC Infect Dis,2012,12(1):224-224.
[11] Chen CS,Yao YC,Lin SC,et al. Retrograde axonal transport:a major transmissionroute of ente rovirus 71 in mice[J]. J Virol,2007,81(17):8996-9003.
[12] Lin TY,Chang LY,Hsia SH,et al.The 1998 enterovirus 71 outbreak in Taiwan:pathogenesis and management[J]. Clin Infect Dis,2002,34(Suppl 2):S52-S57.
[13]赵瑞玲,毛秀娣,张丽锋.利巴韦林在儿科临床应用的安全性评价[J].国际药学研究杂志,2016,43(4):615-620.Zhao RL,Mao XT,Zhang LF. Safety evaluation of ribavirin in clinical practice in paediatrics[J]. J Int Pharmaceut Res,2016,43(4):615-620.
[14]田蕾,李晓艳,徐云霞,等.三白草抗单纯疱疹病毒作用及机制[J].中国中药杂志,2012,37(11):1642-1645.Tian L,Li XY,Xu YX,et al. Eeffect and mechanism of Saururus chinensis against herpes dimplex virus[J].China J Chin Material Medica,2012,37(11):1642-1645.
[15] Khong WX,Foo DG,Trasti SL,et al.Sustained high levels of interleukin-6 contribute to the pathogenesis of enterovirus 71 in a neonate mouse model[J].J Virol,2011,85(7):3067-3076.