【目的】为探讨连翘苷等6种中药成分抗猪传染性胃肠炎病毒(TGEV)的作用机制,明确其体外抗病毒作用效果,为抗TGEV的药物筛选提供依据。【方法】利用动物细胞培养技术,采用MTT比色法和细胞病变(CPE)观察法相结合,测定连翘苷、连翘苷元、...【目的】为探讨连翘苷等6种中药成分抗猪传染性胃肠炎病毒(TGEV)的作用机制,明确其体外抗病毒作用效果,为抗TGEV的药物筛选提供依据。【方法】利用动物细胞培养技术,采用MTT比色法和细胞病变(CPE)观察法相结合,测定连翘苷、连翘苷元、绿原酸、咖啡酸、丁香酚、丹皮酚6种中药成分不同浓度下对猪睾丸细胞(ST)的毒性作用,通过观察细胞CPE情况,确定药物对细胞作用的最大安全浓度;同时测定病毒的TCID_(50),用细胞维持液配成100·TCID_(50)病毒悬液备用;将6种药物在最大安全浓度范围内连续2倍倍比稀释后,分别采用先感染病毒后加药、先加药后感染病毒、药与病毒混合后感染细胞3种不同作用方式进行体外增殖抑制试验,各试验组均设置正常细胞对照组和病毒对照组,每组设置3个重复,通过酶标仪测得630 nm处OD值,计算出不同作用方式下中药成分分别对TGEV的抑制率,筛选出抗TGEV活性较好的中药成分,并分别记录抗病毒效果最佳时药物的浓度;6种中药成分与病毒作用后,测定TGEV的病毒滴度并进行比较分析、RT-PCR鉴定各个药物单体对病毒RNA合成的抑制情况,进一步明确各中药成分对TGEV的抑制作用。【结果】连翘苷、连翘苷元、绿原酸、咖啡酸、丁香酚、丹皮酚的最大安全浓度分别为320、200、80、125、100、200μmol·L^(-1);抗病毒效果最佳时药物的浓度分别为:160、100、20、62.5、25、100μmol·L^(-1)。根据Karber法计算出初始TGEV的TCID_(50)为10^(-6.25)/0.1 m L;6种中药成分对TGEV在ST细胞上均有一定的增殖抑制作用,其中咖啡酸浓度为62.5μmol·L^(-1)时与100·TCID_(50)病毒混合作用后的病毒增殖抑制效果最好,相互作用72 h时,细胞形态依然能够保持圆滑、无固缩、完整,且细胞间轮廓清晰,仅有少量细胞脱落、死亡;此时测上清病毒的TCID_(50)为10^(-3.75)/0.1 m L,结果显示咖啡酸组病毒含量比病�展开更多
Objective：To explore the potential effects of berberine on influenza virus infection both in vitro and in vivo.Methods：In vitro anti-influenza virus assays were performed by cytopathogenic effect and neuraminidase a...Objective：To explore the potential effects of berberine on influenza virus infection both in vitro and in vivo.Methods：In vitro anti-influenza virus assays were performed by cytopathogenic effect and neuraminidase assays in Madin Darby canine kidney cells.In vivo anti-influenza virus assays were performed on the viral pneumonia model of mice.The numbers of mice that died within day 2 to day 14 postinfection were recorded to calculate the mortality.On days 2,4,and 6,the viral titers in the lungs were determined by hemagglutination assay;hematoxylin/eosin staining was used to assess the pathogenic changes of lung tissues; the concentrations of tumor necrosis factor-alpha（TNF-α） and monocyte specific chemoattractant molecule （MCP-1） were measured by radio immunoassay or enzyme-linked immunosorbent assay;the concentrations of nitric oxide（NO） and inducible nitric oxide synthetase（iNOS） were detected by colorimetric method;reverse transcription polymerase chain reaction was used to detect the mRNA level of TNF-αand MCP-1.Results： Berberine showed inhibitory effects on cytopathogenic effects and neuraminidase activity of virus,with the therapeutic index 9.69.In vivo,berberine decreased mice mortality from 90%to 55%,reduced virus titers in the lungs on day 2 postinfection（P0.05）.The lung histology scores were 1.50±0.67,4.50±1.00,and 5.50±1.00 in the berberine group on days 2,4,and 6,respectively,which were significantly reduced compared to 2.17±0.22,6.83±0.44,and 8.50±0.33 in the infected group（P0.05）.The productions of NO and iNOS were repressed by berberine compared with those in the infected group（P0.01）.The transcription and expression of TNF-αwere inhibited by berberine on day 4（P0.01） and day 6（P0.05）,and those of MCP-1 were inhibited on day 6（P0.01） compared with the infected group.Conclusions：Berberine exhibited antiviral effects on the influenza virus both in vitro and in vivo.The possible therapeutic mechanism of berberine on influenza-induced viral pneu展开更多
Antiviral effects of a synthetic Aluminum-Magnesium Silicate (AMS) were tested on Fowl Pox Virus (FPV). Five batches of the Nigerian brand of FPV vaccine were used as sources of the virus. The reconstituted vaccines w...Antiviral effects of a synthetic Aluminum-Magnesium Silicate (AMS) were tested on Fowl Pox Virus (FPV). Five batches of the Nigerian brand of FPV vaccine were used as sources of the virus. The reconstituted vaccines were mixed with The Synthetic AMS on equal volume to weight basis and incubated at room temperature for one hour. They were centrifuged for 10 minutes at 2000 revolutions per minute. The incubation and centrifugation were repeated on a portion of each vaccine supernatant. The two sets of supernatants were tested by the Modified Passive Haemagglutination test, for FPV titres. Portions of the vaccines, not incubated with the AMS, were served as controls. Fowl Pox Virus titres of the vaccines increased from a mean of 2.8 ± 1.10 to 11.2 ± 4.38 when incubated with the AMS once. When incubation with the AMS was repeated, the titres reduced (P< 0.05) to zero in each sample.展开更多
[Objective] To investigate the mechanism of compound traditional Chinese medicine （TCM） on Newcastle disease virus （NDV） and to provide a scientific basis for the reasonable usage of antiviral drugs in clinic. [Me...[Objective] To investigate the mechanism of compound traditional Chinese medicine （TCM） on Newcastle disease virus （NDV） and to provide a scientific basis for the reasonable usage of antiviral drugs in clinic. [Method] The compound TCM was composed of Hedyotis diffusa, Lonicera japonica Thunb, Radix astragali and Glycyrrhiza uralensis. Different dilutions of fluid extract were prepared. Its antiviral effects on NDV were observed through three inoculation ways, first, inoculation with the medicine and NDV mixture which had been incubated at 37 ℃; second, incubating chicken embryo fibroblasts （CEF） with the medicine followed by inoculation with NDV; third, inoculation with N DV followed by incubating CEF with the medicine. The A,= was determined by M]-r [ 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide） ~ method. Therapeutic indexes were used to evaluate the antiviral effects. [ Result] The minimum effective concentration of the compound TCM which acted through the three ways was 1.0 × 2^-10 1.0 × 2^-8 and 1.0 × 2^-7 g/ml, respectively. The antiviral effects of the compound TCM were the best through inoculation with the incubated medicine and NDV mixture, followed by the second method and the third method. [ Conclusion] The compound TCM can not only kill NDV directly in vitro but also inhibit viral propagation.展开更多
文摘【目的】为探讨连翘苷等6种中药成分抗猪传染性胃肠炎病毒(TGEV)的作用机制,明确其体外抗病毒作用效果,为抗TGEV的药物筛选提供依据。【方法】利用动物细胞培养技术,采用MTT比色法和细胞病变(CPE)观察法相结合,测定连翘苷、连翘苷元、绿原酸、咖啡酸、丁香酚、丹皮酚6种中药成分不同浓度下对猪睾丸细胞(ST)的毒性作用,通过观察细胞CPE情况,确定药物对细胞作用的最大安全浓度;同时测定病毒的TCID_(50),用细胞维持液配成100·TCID_(50)病毒悬液备用;将6种药物在最大安全浓度范围内连续2倍倍比稀释后,分别采用先感染病毒后加药、先加药后感染病毒、药与病毒混合后感染细胞3种不同作用方式进行体外增殖抑制试验,各试验组均设置正常细胞对照组和病毒对照组,每组设置3个重复,通过酶标仪测得630 nm处OD值,计算出不同作用方式下中药成分分别对TGEV的抑制率,筛选出抗TGEV活性较好的中药成分,并分别记录抗病毒效果最佳时药物的浓度;6种中药成分与病毒作用后,测定TGEV的病毒滴度并进行比较分析、RT-PCR鉴定各个药物单体对病毒RNA合成的抑制情况,进一步明确各中药成分对TGEV的抑制作用。【结果】连翘苷、连翘苷元、绿原酸、咖啡酸、丁香酚、丹皮酚的最大安全浓度分别为320、200、80、125、100、200μmol·L^(-1);抗病毒效果最佳时药物的浓度分别为:160、100、20、62.5、25、100μmol·L^(-1)。根据Karber法计算出初始TGEV的TCID_(50)为10^(-6.25)/0.1 m L;6种中药成分对TGEV在ST细胞上均有一定的增殖抑制作用,其中咖啡酸浓度为62.5μmol·L^(-1)时与100·TCID_(50)病毒混合作用后的病毒增殖抑制效果最好,相互作用72 h时,细胞形态依然能够保持圆滑、无固缩、完整,且细胞间轮廓清晰,仅有少量细胞脱落、死亡;此时测上清病毒的TCID_(50)为10^(-3.75)/0.1 m L,结果显示咖啡酸组病毒含量比病�
基金Supported by the National Natural Science Foundation of China (No.30772871)
文摘Objective：To explore the potential effects of berberine on influenza virus infection both in vitro and in vivo.Methods：In vitro anti-influenza virus assays were performed by cytopathogenic effect and neuraminidase assays in Madin Darby canine kidney cells.In vivo anti-influenza virus assays were performed on the viral pneumonia model of mice.The numbers of mice that died within day 2 to day 14 postinfection were recorded to calculate the mortality.On days 2,4,and 6,the viral titers in the lungs were determined by hemagglutination assay;hematoxylin/eosin staining was used to assess the pathogenic changes of lung tissues; the concentrations of tumor necrosis factor-alpha（TNF-α） and monocyte specific chemoattractant molecule （MCP-1） were measured by radio immunoassay or enzyme-linked immunosorbent assay;the concentrations of nitric oxide（NO） and inducible nitric oxide synthetase（iNOS） were detected by colorimetric method;reverse transcription polymerase chain reaction was used to detect the mRNA level of TNF-αand MCP-1.Results： Berberine showed inhibitory effects on cytopathogenic effects and neuraminidase activity of virus,with the therapeutic index 9.69.In vivo,berberine decreased mice mortality from 90%to 55%,reduced virus titers in the lungs on day 2 postinfection（P0.05）.The lung histology scores were 1.50±0.67,4.50±1.00,and 5.50±1.00 in the berberine group on days 2,4,and 6,respectively,which were significantly reduced compared to 2.17±0.22,6.83±0.44,and 8.50±0.33 in the infected group（P0.05）.The productions of NO and iNOS were repressed by berberine compared with those in the infected group（P0.01）.The transcription and expression of TNF-αwere inhibited by berberine on day 4（P0.01） and day 6（P0.05）,and those of MCP-1 were inhibited on day 6（P0.01） compared with the infected group.Conclusions：Berberine exhibited antiviral effects on the influenza virus both in vitro and in vivo.The possible therapeutic mechanism of berberine on influenza-induced viral pneu
文摘Antiviral effects of a synthetic Aluminum-Magnesium Silicate (AMS) were tested on Fowl Pox Virus (FPV). Five batches of the Nigerian brand of FPV vaccine were used as sources of the virus. The reconstituted vaccines were mixed with The Synthetic AMS on equal volume to weight basis and incubated at room temperature for one hour. They were centrifuged for 10 minutes at 2000 revolutions per minute. The incubation and centrifugation were repeated on a portion of each vaccine supernatant. The two sets of supernatants were tested by the Modified Passive Haemagglutination test, for FPV titres. Portions of the vaccines, not incubated with the AMS, were served as controls. Fowl Pox Virus titres of the vaccines increased from a mean of 2.8 ± 1.10 to 11.2 ± 4.38 when incubated with the AMS once. When incubation with the AMS was repeated, the titres reduced (P< 0.05) to zero in each sample.
文摘[Objective] To investigate the mechanism of compound traditional Chinese medicine （TCM） on Newcastle disease virus （NDV） and to provide a scientific basis for the reasonable usage of antiviral drugs in clinic. [Method] The compound TCM was composed of Hedyotis diffusa, Lonicera japonica Thunb, Radix astragali and Glycyrrhiza uralensis. Different dilutions of fluid extract were prepared. Its antiviral effects on NDV were observed through three inoculation ways, first, inoculation with the medicine and NDV mixture which had been incubated at 37 ℃; second, incubating chicken embryo fibroblasts （CEF） with the medicine followed by inoculation with NDV; third, inoculation with N DV followed by incubating CEF with the medicine. The A,= was determined by M]-r [ 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide） ~ method. Therapeutic indexes were used to evaluate the antiviral effects. [ Result] The minimum effective concentration of the compound TCM which acted through the three ways was 1.0 × 2^-10 1.0 × 2^-8 and 1.0 × 2^-7 g/ml, respectively. The antiviral effects of the compound TCM were the best through inoculation with the incubated medicine and NDV mixture, followed by the second method and the third method. [ Conclusion] The compound TCM can not only kill NDV directly in vitro but also inhibit viral propagation.
