金银花-连翘药对抗肠道病毒71型的初步探究
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摘要
目的采用体内体外的方法初步探讨金银花-连翘(以下简称"银翘")药对提取物对肠道病毒71型的抑制作用。方法采用MTT法检测"银翘"药对提取物对Hela细胞的毒性以及抗EV71作用;采用腹腔注射20μl EV71建立感染模型,观察小鼠的一般体征,并在光学显微镜下观察心脏组织病理学变化。结果 "银翘"药对提取物对Hela细胞的半数有毒浓度(TC_(50))为123.3 mg/ml,最大无毒浓度(TC_0)为15.6 mg/ml(按生药计)。无论同时加入药物和病毒,还是药物在病毒感染后加入均能够有效抑制病毒,并呈现出一定程度上的剂量依赖关系。体内抗病毒研究显示,中、高剂量组的"银翘"药对提取物均能显著改善小鼠的不良症状和心脏组织的病理变化,效果接近利巴韦林。结论 "银翘"药对提取物能有效地抑制EV71,为其临床应用提供理论依据。
Objective To investigate the antiviral activity of Flos Lonicerae-Fructus Forsythiae herb couples(F-FHC) against enterovirus 71(EV71) in vitro and in vivo.Methods The cytotoxicity and antiviral effect of F-FHC on EV71 infected Hela cells were detected by MTT assay.An infection model was established by intraperitoneal injection of 20 μl EV71.The general characteristics of mice and cardiac pathological morphology were observed under a light microscope.Results The TC_(50) of F-FHC on Hela cells was 123.3 mg/ml,and TC_0 was 15.6 mg/ml.Different concentrations of F-FHC could effectively inhibit EV71 after the EV71 infection or directly mixed with EV71,and showed certain dose-dependent relationship.In vivo antiviral studies showed both medium and high doses of F-FHC could significantly improve the adverse symptoms and cardiac pathological changes,and the effect was similar to that of ribavirin.Conclusion The F-FHC could effectively inhibit EV71,it can provide theoretical basis for its clinical application.
Objective To investigate the antiviral activity of Flos Lonicerae-Fructus Forsythiae herb couples(F-FHC) against enterovirus 71(EV71) in vitro and in vivo.Methods The cytotoxicity and antiviral effect of F-FHC on EV71 infected Hela cells were detected by MTT assay.An infection model was established by intraperitoneal injection of 20 μl EV71.The general characteristics of mice and cardiac pathological morphology were observed under a light microscope.Results The TC_(50) of F-FHC on Hela cells was 123.3 mg/ml,and TC_0 was 15.6 mg/ml.Different concentrations of F-FHC could effectively inhibit EV71 after the EV71 infection or directly mixed with EV71,and showed certain dose-dependent relationship.In vivo antiviral studies showed both medium and high doses of F-FHC could significantly improve the adverse symptoms and cardiac pathological changes,and the effect was similar to that of ribavirin.Conclusion The F-FHC could effectively inhibit EV71,it can provide theoretical basis for its clinical application.
引文
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[9] 陈丽娟,李仁清,张合润,等.EV71所致手足口病患儿及其密切接触儿童带毒状况的追踪观察[J].国际病毒学杂志,2011,18(5):134-138.
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[12] 熊田甜,孟瑜,余光清,等.EV-A71型手足口病重症病例发病危险因素[J].中国热带医学,2018,18(3):242-244,255.
[13] Drake JW,Holland JJ.Mutation rates among RNA viruses [J].Proceedings of the National Academy of Sciences of the United States of America,1999,96(24):13910-13913.
[14] 王剑,侯林,陈亚乔,等.黄芩提取物体外抗病毒药效学研究 [J].中成药,2017,39(9):1924-1927.
[15] Yeh CF,Wang KC,Lu CY,et al.Yakammaoto inhibits enterovirus 71 infection by reducing viral attachment,internalization,replication,and translation [J].Kaohsiung J Med Sci,2015,31:293-302.
[16] 张迎秋,杨怀义,杨倬,等.EV71 感染抗病毒药物及疫苗研究进展 [J].微生物学通报,2012,39(4):544-552.
[17] 薛白琳,姚志红,于荣敏.中药抗 EV71 病毒活性及其在治疗手足口病中的应用 [J].中国中药杂志,2011,36(23):3366-3370.
[18] 李立,杜雅薇,寇爽,等.金银花、连翘对甲型 H1N1 流感免疫调节通路影响的生物信息学分析[J].中国实验方剂学杂志,2017,23(10):201-204.
[19] 吴立夫,张向鹏.金银花连翘提取物对大肠杆菌热敏肠毒素的拮抗作用[J].畜牧兽医学报,1996,26(5):475-480.
[20] 夏伯候,王智民,林丽美,等.银翘药对的药效学研究进展[J].中国实验方剂学杂志,2009,15(3):80-82.
[21] 张莉,李园园,史梅,等.虎杖苷与白藜芦醇体外抗肠道病毒 71 型的初步研究 [J].临床检验杂志,2014,32(10):779-783.
[2] Xiang L,Yuanyuan L,Tingting W,et al.The antiviral effect of baicalin on enterovirus 71 in vitro [J].Viruses,2015,7(8):4756-4771.
[3] 吴嘉瑞,金燕萍,王凯欢,等.基于网络药理学的“金银花-连翘”药对作用机制分析 [J].中国实验方剂学杂志,2017,23(5):179-183.
[4] 罗旋,沈霞,胡本祥.基于分子对接方法进行金银花、连翘抗流感病毒 H7N9 的研究 [J].现代中医药,2016,36(2):75-78.
[5] 周伟.基于“银翘”药对的中药制剂生物有效性综合评价体系的构建与应用[D].南京:南京中医药学,2014.
[6] Ying Q,Lexun L,Yang C,et al.Curcumin inhibits the replication of enterovirus 71 in vitro [J].Acta Pharmaceutica Sinica B,2014,4(4):284-294.
[7] 张霞,任秋生,王海鹏,等.红景天苷体外抗 EV71 病毒的作用 [J].湖北农业科学,2013,52(15):3605-3608.
[8] 曹蓉.甘露消毒丹对感染小鼠血清酶活性的影响研究[M].长沙:湖南中医药大学,2015.
[9] 陈丽娟,李仁清,张合润,等.EV71所致手足口病患儿及其密切接触儿童带毒状况的追踪观察[J].国际病毒学杂志,2011,18(5):134-138.
[10] 刘金亭,刘力建.EV71-IgM检测在手足口病诊断中的价值[J].中国社区医师,2016,32(8):133,135.
[11] 王静凛.金银清瘟汤治疗EV71型手足口病肺脾湿热证临床研究[D].济南:山东中医药大学,2011.
[12] 熊田甜,孟瑜,余光清,等.EV-A71型手足口病重症病例发病危险因素[J].中国热带医学,2018,18(3):242-244,255.
[13] Drake JW,Holland JJ.Mutation rates among RNA viruses [J].Proceedings of the National Academy of Sciences of the United States of America,1999,96(24):13910-13913.
[14] 王剑,侯林,陈亚乔,等.黄芩提取物体外抗病毒药效学研究 [J].中成药,2017,39(9):1924-1927.
[15] Yeh CF,Wang KC,Lu CY,et al.Yakammaoto inhibits enterovirus 71 infection by reducing viral attachment,internalization,replication,and translation [J].Kaohsiung J Med Sci,2015,31:293-302.
[16] 张迎秋,杨怀义,杨倬,等.EV71 感染抗病毒药物及疫苗研究进展 [J].微生物学通报,2012,39(4):544-552.
[17] 薛白琳,姚志红,于荣敏.中药抗 EV71 病毒活性及其在治疗手足口病中的应用 [J].中国中药杂志,2011,36(23):3366-3370.
[18] 李立,杜雅薇,寇爽,等.金银花、连翘对甲型 H1N1 流感免疫调节通路影响的生物信息学分析[J].中国实验方剂学杂志,2017,23(10):201-204.
[19] 吴立夫,张向鹏.金银花连翘提取物对大肠杆菌热敏肠毒素的拮抗作用[J].畜牧兽医学报,1996,26(5):475-480.
[20] 夏伯候,王智民,林丽美,等.银翘药对的药效学研究进展[J].中国实验方剂学杂志,2009,15(3):80-82.
[21] 张莉,李园园,史梅,等.虎杖苷与白藜芦醇体外抗肠道病毒 71 型的初步研究 [J].临床检验杂志,2014,32(10):779-783.