10种中成药体外抗流感病毒活性研究

流感是一种高发病率和高死亡率的上呼吸道传染病, 我国是流感高发区。国际上预防与治疗流感的主要措施是疫苗接种和抗病毒化学药物治疗。除此之外我国临床上还利用传统中药进行抗流感病毒治疗。本研究采用细胞病变 (CPE) 法, 对10种中成药进行体外抗流感病毒活性检测。厂家H生产的清热解毒口服液体外对A/广东罗湖/219/2006 (H1N1) 具有较强的抑制活性; 银黄口服液对A/汉防/359/95和A/粤防/243/72, 厂家G生产的清开灵口服液对A/济防/15/90, 厂家H生产的清热解毒口服液对A/济防/15/90、A/粤防/243/72和B/深圳/155/2005具有较强的抑制活性。     

双黄连口服液抗呼吸道合胞病毒的实验研究

目的:评价双黄连口服液抗呼吸道合胞病毒(RSV)的活性。方法:以病毒唑为对照,通过体外观察病毒致细胞病变效应(CPE)、病毒滴度、抗病毒指数,在体内观察其对染毒小鼠的死亡保护作用,以及对小鼠心、脑、肺内病毒增殖的影响,从而判定双黄连口服液抗RSV作用。结果:双黄连口服液能抑制RSV的增殖,其有效浓度为12ug/mL,并显示出病毒的感染性滴度随药物浓度的增加而下降;治疗RSV感染鼠结果发现,双黄连口服液对RSV感染鼠有保护作用,6.7g.k-g 1.d-1时,RSV感染鼠的存活率为75%,平均存活天数为(12.9±2.8)d,随着药物剂量增加,疗效增强;该口服液还能降低组织内病毒滴度,阻止体内病毒复制,抗RSV作用类似于同剂量的病毒唑。结论:双黄连口服液是一种有效的抗RSV中药复方制剂。     

腔肠素抗流感病毒研究

目的 对海洋来源的化合物腔肠素进行体外抗流感病毒活性研究。方法 细胞病变效应(CPE)抑制实验评价腔肠素对不同流感病毒株的抗病毒作用。CPE实验检测腔肠素细胞毒性和抗病毒作用方式并结合血凝抑制实验、神经氨酸酶(NA)活性实验及微基因组实验初步探究腔肠素的作用靶点。结果 腔肠素体外对不同流感病毒株均有良好抗病毒作用,其中对PR8的抑制作用最强,IC₅₀为5.0μmol·L^-1且细胞毒较小,CC₅₀为144.1μmol·L^-1。通过不同作用方式以及不同作用时间的实验显示,腔肠素预先处理流感病毒以及在病毒感染后加入均有良好抗病毒作用,并且在病毒吸附后0～3 h的抑制效果较好。用腔肠素预处理病毒后可以呈剂量依赖性地抑制血凝素(HA)的活力。故腔肠素可能是通过与病毒HA相互作用来阻断流感病毒的进入,其作用位点可能为HA蛋白HA2链的72、74位谷氨酸。结论 腔肠素具有良好的体外抗甲型流感病毒效果,为海洋来源的小分子化合物的开发与改造提供了借鉴,为寻找新型抗流感病毒药物提供新思路。     

复方抗病毒制剂体外抗流感病毒和呼吸道合胞病毒作用及毒性研究

目的 测定复方抗病毒制剂体外对流感病毒和呼吸道合胞病毒的抗病毒作用. 方法 用鸡红细胞血凝素滴定法测定不同浓度复方抗病毒制剂在狗肾传代细胞（MDCK）中对流感病毒的抑制作用;用中和实验测定复方抗病毒制剂在Hep 2细胞中对呼吸道合胞病毒的活性. 结果 复方抗病毒制剂浓度＜1.5 mg•mL^-1时对MDCK细胞及Hep 2细胞无明显毒性. 浓度＞0.15 mg•mL^-1能抑制流感病毒的血凝活性和呼吸道合胞病毒对靶细胞的攻击作用. 结论 复方抗病毒制剂体外对流感病毒和呼吸道合胞病毒具有较强的抑制作用,且有较大的安全性.     

双黄连口服液中黄芩甙含量测定方法比较

双黄连口服液中黄芩甙含量测定方法比较李及鲁静１（成都市药品检验所６１００００；１中国药品生物制品检定所）双黄连口服液为临床常用的治疗呼吸道疾病的中成药，具有辛凉解表、清热解毒作用。目前已有多家药厂生产，质量标准收载于卫生部部颁标准〔１〕，其含量测定项...     

扎那米韦等对流感病毒神经氨酸酶的抑制作用

目的:观察扎那米韦等6种临床抗流行性感冒或抗病毒上呼吸道感染药物对流感病毒神经氨酸酶(neuraminidase ,NA)的抑制作用.方法:应用荧光测定法进行体外酶活性测定.结果:扎那米韦对我国分离的3株流感病毒粤防72 243、济防90 15、四川2000 38的神经氨酸酶均有抑制作用,其半效抑制浓度(IC50)分别为0.40,0.23,2.57 nmol•L 1.金刚烷胺、金刚乙胺、利巴韦林(RBV)、α 干扰素(α IFN)和中药抗病毒口服液对流感病毒神经氨酸酶无抑制作用.结论:扎那米韦对我国流感病毒神经氨酸酶具有抑制活性.     

桑白皮抗病毒有效成分的提取分离及体外抗病毒活性研究

目的研究桑白皮中化学成分的抗病毒作用。方法利用细胞病变方法进行体外抗病毒筛选试验。结果桑白皮中分离得到的化合物1和化合物3在体外有较好的抑制副流感病毒、流感病毒的致病作用;化合物1具有抗呼吸道合胞病毒作用,延缓腺病毒Ⅲ、HSV Ⅰ致病作用;化合物2和化合物5也有部分抗病毒作用;化合物4能抑制腺病毒Ⅲ、柯萨奇病毒B3 、HSV Ⅰ、副流感病毒的致细胞病变作用。结论桑白皮提取物对呼吸道常见病毒有抑制作用。     

双黄连颗粒体外抗呼吸道合胞病毒作用的实验研究

目的研究双黄连颗粒体外抑制呼吸道合胞病毒(RSV)的作用。方法通过细胞培养,观察细胞病变(CPE),观察双黄连颗粒在人喉癌上皮细胞株(Hep-2)中抑制RSV的作用。结果双黄连颗粒能抑制RSV复制,具有明显的抗病毒作用。结论双黄连颗粒具有抑制RSV在Hep-2细胞内复制的作用。     

黄芩苷与利巴韦林联用体内外抗流感病毒作用

目的评价黄芩苷与利巴韦林联用体内外抗流感病毒作用,为其临床上联用提供实验依据。方法 CPE法和MTT法测定药物单用及联用对感染3种不同亚型流感病毒MD-CK细胞的保护效果,分别应用合并指数法(combination in-dex,CI)和MacSynergyⅡ分析法,分析黄芩苷与利巴韦林体外联用的相互作用。神经氨酸酶(NA)抑制试验测定黄芩苷对流感病毒NA活性的抑制作用。应用H1N1亚型流感病毒(FM1株)人工感染BALB/c小鼠模型,评价黄芩苷与利巴韦林联用对感染小鼠的保护作用。结果体外黄芩苷与利巴韦林联用对H1N1和H5N1病毒的抑制表现为相加作用,对H9N2病毒的抑制表现为协同作用。黄芩苷对3种亚型流感病毒NA的半数抑制浓度(IC50)在0.29～0.49 mmol.L-1范围。利巴韦林与黄芩苷联合给药与单独给药相比,提高感染小鼠的存活率,延长其存活时间。结论黄芩苷对流感病毒NA活性具有抑制作用。黄芩苷与利巴韦林在体外联用,对不同亚型流感病毒的抑制表现为相加或协同作用;二者联用对感染流感小鼠的保护效果明显优于单用。     

双黄连与阿米卡星注射液存在配伍禁忌

双黄连口服液是近年来广泛应用于临床的纯中药复方制剂,为中药金银花、连翘、黄芩等提取有效成份制成,具有抗菌消炎,清热解毒之功效。临床上双黄连口服液常与抗生素配合使用,治疗流行性感冒及上呼吸道感染、病毒性肺炎,以及球菌的各种感     

海洋来源的吲哚生物碱衍生物抗流感病毒作用研究

目的 对海洋来源的吲哚生物碱类化合物HDYL-GQQ-1932进行体外抗流感病毒活性研究。方法 CPE抑制实验测定HDYL-GQQ-1932对多种病毒株的增殖抑制作用。CPE实验结合血凝抑制和神经氨酸酶活性实验探究HDYL-GQQ-1932 作用病毒入侵细胞的方式以及细胞毒性,并且初步探究HDYL-GQQ-1932的作用靶点。 结果 HDYL-GQQ-1932体外对多种流感病毒株均有增殖抑制作用,其中对H1N1的抑制作用最强,IC₅₀为26.1μM ,且无明显的细胞毒性,CC₅₀为3313.2μM。通过不同作用方式以及不同作用时间的实验显示,感染后加入 HDYL-GQQ-1932能够很好的抑制病毒增值,且用HDYL-GQQ-1932预处理细胞后可明显降低病毒增殖。此外结果显示HDYL-GQQ-1932在病毒吸附后6-9 h有很好的抑制作用,且剂量依赖性地抑制神经氨酸酶(NA)的活力。故 HDYL-GQQ-1932可能是通过结合病毒神经氨酸酶并抑制其活性来阻断IAV的释放过程。结论 HDYL-GQQ-1932体外对甲型流感具有较好的抑制效果。本研究为海洋来源的该类小分子化合物的药物开发与理性改造提供了思路与方向。     

In vitro anti-influenza virus and anti-inflammatory activities of theaflavin derivatives

The theaflavins fraction (TF80%, with a purity of 80%) and three theaflavin (TF) derivatives from black tea have been found to exhibit potent inhibitory effects against influenza virus in vitro. They were evaluated with a neuraminidase (NA) activity assay, a hemagglutination (HA) inhibition assay, a real-time quantitative PCR (qPCR) assay for gene expression of hemagglutinin (HA) and a cytopathic effect (CPE) reduction assay. The experimental results showed that they all exerted significant inhibitory effects on the NA of three different subtypes of influenza virus strains [A/PR/8/34(H1N1), A/Sydney/5/97(H3N2) and B/Jiangsu/10/2003] with 50% inhibitory concentration (IC(50)) values ranging from 9.27 to 36.55 μg/mL, and they also displayed an inhibitory effect on HA; these inhibitory effects might constitute two major mechanisms of their antiviral activity. Time-of-addition studies demonstrated that TF derivatives might have a direct effect on viral particle infectivity, which was consistent with the inhibitory effect on HA. Subsequently, the inhibitory effect of TF derivatives on the replication of the viral HA gene as assayed by qPCR and on the nuclear localization of the influenza virus vRNP further demonstrated that they may primarily act during the early stage of infection. Interestingly, besides the activity against functional viral proteins, TF derivatives also decreased the expression level of the inflammatory cytokine IL-6 during viral infection, expression of which may result in serious tissue injury and apoptosis. Our results indicated that TF derivatives are potential compounds with anti-influenza viral replication and anti-inflammatory properties. These findings will provide important information for new drug design and development for the treatment of influenza virus infection.     

In vitro Antiviral Effects and 3D QSAR Study of Resveratrol Derivatives as Potent Inhibitors of Influenza H1N1 Neuraminidase

The anti-influenza virus activities of 50 resveratrol (RV: 3, 5, 4′-trihydroxy-trans-stilbene) derivatives were evaluated using a neuraminidase (NA) activity assay. The results showed that 35 compounds exerted an inhibitory effect on the NA activity of the influenza virus strain A/PR/8/34 (H1N1) with 50% inhibitory concentration (IC₅₀) values ranging from 3.56 to 186.1 μm . Next, the 35 RV derivatives were used to develop 3D quantitative structure–activity relationship (3D QSAR) models for understanding the chemical–biological interactions governing their activities against NA. The comparative molecular field analysis (CoMFA r² = 0.973, q² = 0.620, q_test² = 0.661) and the comparative molecular similarity indices analysis (CoMSIA r² = 0.956, q² = 0.610, q_test² = 0.531) were applied. Afterward, molecular docking was performed to study the molecular interactions between the RV derivatives and NA. Finally, a cytopathic effect (CPE) reduction assay was used to evaluate the antiviral effects of the RV derivatives in vitro. Time-of-addition studies demonstrated that the RV derivatives might have a direct effect on viral particle infectivity. Our results indicate that the RV derivatives are potentially useful antiviral compounds for new drug design and development for influenza treatment.     

Anti-influenza virus activities of flavonoids from the medicinal plant Elsholtzia rugulosa

ELSHOLTZIA RUGULOSA (Lamiaceae), a common Chinese herb, is widely used in the treatment of cold and fever. In order to elucidate the action mechanism and the active principles from the plant against anti-influenza virus, the influenza virus neuraminidase (NA) activity assay and IN VITRO antiviral activity assay were established, and the isolation of the active principles was guided by NA activity. Finally, 5 active constituents were obtained, namely apigenin ( 1), luteolin ( 2), apiin ( 3), galuteolin ( 4) and luteolin 3'-glucuronyl acid methyl ester ( 5), respectively. They all belong to the flavonoids. The IN VITRO antiviral assay using a cytopathic effect (CPE) reduction method showed that they all possessed anti-influenza virus activity. Among them, apigenin and luteolin exhibited the highest activities against influenza virus (H3N2) with IC (50) values of 1.43 microg/mL and 2.06 microg/mL, respectively. The structure-activity relationship (SAR) of these flavonoids with different chemical structures and their anti-influenza virus activities was addressed in this study.     

Patchouli alcohol: in vitro direct anti-influenza virus sesquiterpene in <Emphasis Type="Italic">Pogostemon cablin</Emphasis> Benth.

During the screening of anti-influenza virus substances from traditional herbal medicines, the methanol extract from the leaves of Pogostemon cablin Benth. showed potent in vitro antiviral activity (99.8% inhibition at a concentration of 10 μg/mL) against influenza virus A/PR/8/34 (H1N1). The anti-influenza virus principle was isolated from the hexane-soluble fraction, through solvent fractionation, repeated silica gel column chromatography, and reversed-phase HPLC. The major active principle was a volatile substance that was identified as a sesquiterpene, patchouli alcohol (1), on the basis of its spectral analyses. When anti-influenza virus activity against A/PR/8/34 was evaluated by the plaque forming assay, patchouli alcohol reduced the number of plaques by 75% at 2 μg/mL and 89% at 10 μg/mL. Patchouli alcohol showed dose-dependent anti-influenza virus activity, and its IC₅₀ value was estimated to be 2.635 μM. Although 11 different sesquiterpenes were tested for antiviral activity against influenza virus A/PR/8/34, no or negligible activity was observed except for patchouli alcohol. Patchouli alcohol did not show anti-influenza virus activity against A/Guizhou/54/89 (H3N2), but showed weak activity against B/Ibaraki/2/85 (IC₅₀ = 40.82 μM). Patchouli alcohol did not show inhibitory activity against influenza virus neuraminidase.     

利巴韦林注射液体内外抗流感病毒作用研究

利巴韦林具有广谱抗病毒活性, 对多种DNA或RNA病毒体内外都有抑制作用。本文对利巴韦林注射液的体内外抗流感病毒活性进行研究, 体外实验采用细胞病变 (CPE) 法研究了利巴韦林注射液对流感病毒甲乙型的活性, 体内实验采用小鼠模型研究了利巴韦林注射液对流感病毒A/FM/1/47 (H1N1) 鼠肺适应株感染小鼠的保护作用。结果显示, 在体外实验中, 利巴韦林注射液对所检测的7种病毒株均有显著的抑制活性; 在体内实验中, 利巴韦林注射液可以显著提高感染小鼠的存活率和平均生活日, 显著改善感染小鼠的肺病变和肺指数。     

Inhibition of influenza virus internalization by (−)-epigallocatechin-3-gallate

(−)-Epigallocatechin-3-gallate (EGCG), one of the major flavonoid components of green tea, is known to have a broad antiviral activity against several enveloped viruses, including the influenza virus. However, its mode of action and the mechanism that allows it to target influenza virus molecules have not been fully elucidated. Thus, this study investigated the molecular mechanism by which EGCG suppresses influenza virus infections. EGCG was found to block an early step in the influenza viral life cycle, but it did not affect viral adsorption to target cells or viral RNA replication. However, EGCG inhibited hemifusion events between virus particles and the cellular membrane by reducing the viral membrane integrity, thereby resulting in the loss of the cell penetration capacity of the influenza virus. EGCG also marginally suppressed the viral and nonviral neuraminidase (NA) activity in an enzyme-based assay system. In conclusion, it is suggested that the anti-influenza viral efficacy of EGCG is attributable to damage to the physical properties of the viral envelope and partial inhibition of the NA surface glycoprotein. These results may facilitate future investigations of the antiviral activity of EGCG against other enveloped viruses as well as influenza virus.     

芩连胶囊抗流感病毒活性的体内外实验

目的研究芩连胶囊在体内外的抗流感病毒活性。方法采用MTF法检测芩连胶囊对体外模型NDCK细胞的毒性浓度。采用CPE法测定芩连胶囊体外抑制流感有效浓度。动物模型为昆明种小鼠,口服给药,qd,共给药6 d。通过观察小鼠病死率、生命延长率及肺病变程度指标判定其抗流感病毒作用。结果芩连胶囊在体外能有效抑制流感A3病毒CPE的产生;口服芩连胶囊,能减少流感病毒FM1感染小鼠的病死率,延长存活时间,减轻小鼠肺组织病变程度。结论芩连胶囊在体内外均有良好的抗流感病毒活性。     

我院2004年～2005年门诊中成药应用分析

目的：分析门诊中成药在我院临床的应用情况,为临床合理用药提供参考。方法：利用医院数据库,统计2004年～2005年使用数据,通过用药金额、用药频度等情况分析药品应用情况。结果及结论：用药情况符合我院以治疗肝炎为主的传染病专科医院的特点,用药以口服剂型为主,注射剂呈上升趋势,用药品种较为集中,且金额低的药物使用多,用药基本合理。护肝宁片、养血饮口服液、复方鳖甲软肝片、软肝颗粒、亮菌口服液在肝炎治疗中使用广泛。我院自制制剂软肝颗粒有开发的前景。