21-清咽滴丸抗柯萨奇病毒CVB3,CVB5作用的研究

目的:探究清咽滴丸及其六种中药成分(甘草、青黛、冰片、诃子、牛黄和薄荷)提取物对柯萨奇病毒B3、B5的抑制作用。方法:采用MTT法检测活细胞生存率,结合观察CPE,在细胞水平上检验清咽滴丸及其六种成分对CVB3、CVB5增殖的抑制作用。取其有效成分后,进一步用血清药理学的方法,检验经机体吸收后药物是否仍然具有抗病毒的效果。结果:在细胞水平上,清咽滴丸及其六种主要成分都对细胞有明显毒性作用,但只有清咽滴丸、甘草、诃子可以抑制CVB3、CVB5在细胞内的增殖。血清药理学进一步验证清咽滴丸、甘草、诃子对CVB3、CVB5有抑制作用。结论:清咽滴丸对CVB3、CVB5有一定的抗感染能力和治疗效果,其中起主要作用的成分是甘草和诃子,且抑制作用呈一定的浓度、时间依赖性。     

Antiviral Activity of Hederasaponin B from Hedera helix against Enterovirus 71 Subgenotypes C3 and C4a

Enterovirus 71 (EV71) is the predominant cause of hand, foot and mouth disease (HFMD). The antiviral activity of hederasaponin B from Hedera helix against EV71 subgenotypes C3 and C4a was evaluated in vero cells. In the current study, the antiviral activity of hederasaponin B against EV71 C3 and C4a was determined by cytopathic effect (CPE) reduction method and western blot assay. Our results demonstrated that hederasaponin B and 30% ethanol extract of Hedera helix containing hederasaponin B showed significant antiviral activity against EV71 subgenotypes C3 and C4a by reducing the formation of a visible CPE. Hederasaponin B also inhibited the viral VP2 protein expression, suggesting the inhibition of viral capsid protein synthesis.These results suggest that hederasaponin B and Hedera helix extract containing hederasaponin B can be novel drug candidates with broad-spectrum antiviral activity against various subgenotypes of EV71.     

天然(口山)酮化合物抗柯萨奇病毒B_5的作用

目的 研究天然咄酮类化合物抗柯萨奇病毒B_5(CVB_5)的作用及其机理.方法 通过改变给药途径,观察病毒引起的细胞病变效应(CPE),MTT法检测细胞活性,评价酮化合物在Hep-2细胞培养中的抗CVB_5作用.结果 酮具有抗CVB_5的活性,对Hep-2细胞的TC_(50)为54.44 μg·mL~(-1),其EC_(50)分别为26.378、25.813、25.405 μg·mL~(-1),治疗指数(TI)分别为2.064、2.109、2.143;在32 μg·mL~(-1)时,对CVB_5的抑制率达50%.结论 酮化合物通过直接杀灭病毒、阻止病毒的吸附、抑制病毒的生物合成而发挥其抗CVB_5的作用;但需对结构做进一步的修饰,以降低其毒性.     

Antiviral activity of arbidol hydrochloride against herpes simplex virus I in vitro and in vivo

Herpes simplex virus type 1 (HSV-1) causes significant human diseases ranging from skin lesions to encephalitis, especially in neonates and immunocompromised hosts. The discovery of novel anti-HSV-1 drugs with low toxicity is required for public health. Arbidol hydrochloride (ARB) is an indole derivative molecule with broad-spectrum antiviral activity. In this study, the antiviral effects of ARB against HSV-1 infection were evaluated in vitro and in vivo. The results showed that ARB presents significant inhibitory effect on HSV-1 plaque formation and generation of progeny virus, with EC50 values (50% effective concentration) of 5.39?µg/mL (10.49?µM) and 2.26?µg/mL (4.40?µM), respectively. Moreover, time-of-addition and time-of-removal assays further suggested that ARB has viral inhibitory effects when added up to 12?h post-infection (p.i.), which could be further corroborated by determining the expression of viral immediate-early (ICP4, ICP22 and ICP27), early (ICP8 and UL42) and late (gB, gD, gH, VP1/2 and VP16) genes by real-time quantitative PCR as well as the expression of viral protein ICP4 and ICP8 at 6?h and 12?h p.i. Results of the in vivo study showed that ARB could reduce guinea pig skin lesions caused by HSV-1 infection. Conclusively, this report offers new perspectives in the search for therapeutic measures in the treatment of HSV-1 infection.     

清心饮对柯萨奇B3病毒感染心脏微血管内皮细胞的保护作用

目的 研究清心饮对柯萨奇B₃病毒（Coxsackievirus B3,CVB₃）感染心脏微血管内皮细胞（cardiac microvascular endothelial cells,CMVECs）的保护作用。方法 采用RT-PCR检测CVB₃ RNA,应用倒置显微镜观察CMVECs形态,CCK8检测细胞活性,选择清心饮最佳干预时间点和稀释浓度。在最佳条件下通过免疫荧光法、Western blot法检测内皮细胞标志物CD31,间充质细胞标志物α-SMA的表达。结果 CVB₃能在CMVECs中持续性复制,清心饮最佳干预时间点和浓度分别是72 h和20%含药血清。与空白组比较,CVB₃组CMVECs形态固缩、变异,CD31表达减弱,α-SMA表达增强;与空白血清组比较,清心饮血清组CMVECs形态趋于正常,CD31表达增强,α-SMA表达减弱。结论 清心饮可抑制CVB₃感染CMVECs发生内皮细胞转分化,对CMVECs具有保护作用。     

双黄连粉针剂抗柯萨奇病毒B3型效应的实验研究

目的：探讨双黄连在体外抗柯萨奇病毒（ＣＶＢ３）的效应。方法在Ｗｉｓｈ细胞上采用微量细胞病变抑制法。结果（１）３．０ｍｇ／ｍｌ、１．５ｍｇ／ｍｌ双黄连均可直接杀伤１００、１０ＴＣＩＤ５０病毒,且３．０ｍｇ／ｍｌ直接灭活ＣＶＢ３的产应明显。（２）０．５、０．２５、０．１２５ｍｇ／ｍｌ双黄连可分别阻断１００、１０、１ＴＣＩＤ５０病毒吸附细胞。（３）１．５、０．２５、０．２５ｍｇ／ｍｌ双黄连可分别抑制１０     

Antiviral activity of methyl helicterate isolated from Helicteres angustifolia (Sterculiaceae) against hepatitis B virus

The anti-HBV effect of methyl helicterate (MH), a triterpenoid isolated from the Chinese herb Helicteres angustifolia, was explored both in vitro and in vivo. In the HBV-transfected cell line HepG2.2.15, the secretion of HBsAg/HBeAg, the levels of HBV DNA and cccDNA, and the amount of viral RNA were significantly decreased after treatment with MH for 144 h. In addition, MH had no inhibitory effect on the mitochondrial DNA content. In DHBV-infected ducklings, MH significantly reduced the serum DHBV DNA, liver total viral DNA, and cccDNA levels. Furthermore, analysis of the liver pathological changes confirmed the hepatoprotective effect of MH. These results indicate that MH efficiently inhibits HBV replication both in vitro and in vivo and that MH may be a major bioactive ingredient in H. angustifolia.     

Sclerotiorin的胺类衍生物的潜在抗病毒活性及其初步构效关系研究

目的 为了评价天然产物(+)-sclerotiorin及其半合成衍生物的抗病毒活性。方法 通过CPE法评价(+)-Sclerotiorin (1)及其半合成衍生物(2-31)对RSV,EV71,HSV-1,H1N1和Cox-B3的抗病毒活性。结果 化合物3和11显示出对EV71的抗病毒活性,并且具有非常高的选择性指数值(SI值分别为37.8和38.0)。结论 化合物3和11是潜在的抗病毒先导化合物。     

Combination of soluble coxsackievirus-adenovirus receptor and anti-coxsackievirus siRNAs exerts synergistic antiviral activity against coxsackievirus B3

Coxsackievirus B3 (CVB-3) is a major causative agent of chronic heart muscle infections. The present study describes a cell culture system with an ongoing virus infection to evaluate two novel inhibitory strategies, either individually or combined: (1) RNA interference (RNAi) to degrade cytoplasmatic CVB-3 RNA and (2) a vector-delivered soluble variant of the coxsackievirus-adenovirus receptor fused to a human immunoglobulin (sCAR-Fc), which inhibits cellular uptake of CVB-3. Both approaches were capable of inhibiting CVB-3 in persistently infected human myocardial fibroblasts. The antiviral effect of a single treatment lasted for up to one week and could be extended by repeated applications. Each of the single treatments initially reduced the virus titer by approximately 1-log, whereas the combination of both approaches resulted in 4-log inhibition and retained substantial antiviral activity at later time points, when the effect of sCAR-Fc or siRNAs alone had already disappeared. Further analysis revealed that sCAR-Fc protects cells from virus-induced lysis but does not diminish the virus load. Reduction of the virus titer was only achieved with additional destruction of viral RNA by RNAi. Taken together, combination of RNAi and a protein-based antiviral strategy was found to result in a strong synergistic inhibition of an ongoing virus infection.     

苦参系列生物碱体外抗CVB_3病毒活性

目的研究苦参系列生物碱(Sophora flavescens alkaloids)对柯萨奇B3病毒(CVB3)所致的Vero细胞和大鼠原代心肌细胞病变抑制作用、病毒繁殖抑制反应,并测定其对大鼠原代心肌细胞培养上清液中心肌酶(LDH、CK MB)活性的影响。方法在Vero细胞、大鼠原代心肌细胞中加入不同稀释度苦参系列生物碱后,感染CVB3病毒,观察细胞病变,MTT染色比较各组间活细胞数的变化,病毒繁殖抑制实验比较半数组织感染量(TCID50)的差别;紫外分光光度法测定心肌细胞培养液上清中LDH和CK MB活性。结果槐果碱、苦参碱、槐啶碱在Vero细胞和原代乳鼠心肌细胞试验中可有效抑制CVB3病毒引起的细胞病变(P<0.05,P<0.01);槐果碱对病毒的繁殖有抑制作用(对数值升高一个单位),并可降低心肌细胞释放心肌酶(P<0.05,P<0.01)。结论槐果碱、苦参碱、槐啶碱对CVB3病毒感染的Vero细胞和原代乳鼠心肌细胞具有一定保护作用。对细胞病变(CPE)的抑制作用强弱顺序为:槐果碱>苦参碱>槐定碱>氧化槐果碱、苦豆碱、槐胺碱、槐醇碱。槐果碱对CVB3病毒的繁殖有抑制作用,在一定浓度下可降低心肌酶的释放,其作用机制尚需做进一步探讨。     

养心颗粒对柯萨奇病毒B_3感染小鼠心脏内细胞因子的调节作用研究

目的:研究养心颗粒对柯萨奇病毒B3感染小鼠心脏内肿瘤坏死因子(TNF)-α和干扰素(IFN)-γ的调节作用。方法:腹膜下注射柯萨奇病毒B3接种Balb/c小鼠。实验分为对照、黄芪提取物(6 g·kg-1)和养心颗粒高、低剂量(6、3 g·kg-1)组,分别于接种后第7、14、28天采集小鼠心脏标本,以酶联免疫吸附法测定心脏内TNF-α和IFN-γ的含量。结果:与对照组比较,第7天养心颗粒高、低剂量组感染小鼠心脏内IFN-γ水平显著升高,但养心颗粒高剂量组自第14天开始、低剂量组第28天开始,对感染小鼠心脏内TNF-α的释放显著减弱(P<0.05)。结论:养心颗粒对柯萨奇病毒B3感染小鼠心脏细胞因子TNF-α和IFN-γ的调节作用与黄芪提取物相似,并且其在感染后期对TNF-α和IFN-γ的抑制作用可能强于黄芪。     

大蒜多糖体外抗柯萨奇病毒B3作用

目的研究大蒜多糖体外抗柯萨奇病毒B3(Coxsackievirusg roup B type 3,CVB3)作用。方法观察大蒜多糖A,B,C的细胞毒性、对CVB3直接灭活作用、抗CVB3吸附作用及对CVB3生物合成抑制作用。结果大蒜多糖A,B,C对Hep-2细胞的半数中毒浓度(TC50)分别为1000,800,4000μg／mL;均无直接灭活CVB3及抗CVB3吸附作用;除A外,B和C均可剂量依赖性抑制CVB3生物合成。结论大蒜多糖B和C在体外通过抑制CVB3生物合成而发挥抗CVB3的作用。     

Spiramycin and azithromycin,safe for administration to children,exert antiviral activity against enterovirus A71 in vitro and in vivo

Hand-foot-mouth disease (HFMD) is a common viral disease in young children, mainly caused by enterovirus A71 (EV-A71) and coxsackievirus A16 (CV-A16). Specific antiviral agents are not commercially available yet. Here we report that the macrolide antibiotics spiramycin (SPM) and azithromycin (AZM) possess antiviral activities against EV-A71 and CV-A16. SPM significantly reduced EV-A71 RNA and protein levels, most likely through interfering with viral RNA replication. The SPM-resistant EV-A71 variants showed similar resistance to AZM, indicating a similar anti-EV-A71 mechanism by which these two drugs exert their functions. The mutations of these variants were reproducibly mapped to VP1 and 2A, which were confirmed to confer resistance to SPM. Animal experiments showed that AZM possesses stronger anti-infection efficacy than SPM, greatly alleviated the disease symptoms and increased the survival rate in a mouse model severely infected with EV-A71. In all, our work suggests that AZM is a potential treatment option for EV-A71-induced HFMD, whose proved safety for infants and children makes it even more promising.     

Phyllaemblicin B inhibits Coxsackie virus B3 induced apoptosis and myocarditis

Coxsackie virus B3 (CVB3) is believed to be a major contributor to viral myocarditis since virus-associated apoptosis plays a role in the pathogenesis of experimental myocarditis. In this study, we investigated the in vitro and in vivo antiviral activities of Phyllaemblicin B, the main ellagitannin compound isolated from Phyllanthus emblica, a Chinese herb medicine, against CVB3. Herein we report that Phyllaemblicin B inhibited CVB3-mediated cytopathic effects on HeLa cells with an IC₅₀ value of 7.75 ± 0.15 μg/mL. In an in vivo assay, treatment with 12 mg kg⁻¹ d⁻¹ Phyllaemblicin B reduced cardiac CVB3 titers, decreased the activities of LDH and CK in murine serum, and alleviated pathological damages of cardiac muscle in myocarditic mice. Moreover, Phyllaemblicin B clearly inhibited CVB3-associated apoptosis effects both in vitro and in vivo. These results show that Phyllaemblicin B exerts significant antiviral activities against CVB3. Therefore, Phyllaemblicin B may represent a potential therapeutic agent for viral myocarditis.     

广谱抗病毒抗生素17997体内外抗病毒活性研究

从我国云南省土壤中分离到１株放线菌，基们生的抗生素１７９９７具有广谱抗病毒作用在细菌培养内对单纯疱疹病毒１型，单纯疱疹病毒２型，人免疫缺陷病毒１型，水疱性口炎病毒及柯萨奇病毒Ｂ３均有显著抑制活性，ＩＣ５０在μｍｏｌ／Ｌ水平。其在小鼠体内对单纯疱疹病毒１型及２型所致疱疹性脑炎有确切治疗效果，对死亡保护率平均生存日与对照组相比有统计意义的显著差别。     

2—氯—5—（5，6—二氢—2—甲基—1，4—氧硫杂环己二烯—3—甲酰胺基）苯甲酸异丙酯类似物的合成和生物活性

2－氯－5－（5,6－二氢－2－甲基－1,4－氧硫杂环己二烯－3－甲酰胺基）苯甲酸异丙酯（UC84）具有显著的抗病毒活性。合成了该化合物并以其为先导物模型制备了十一个结构未经报道的类似物,活性筛选试验表明该化合物有明显的抗乙肝病毒（HBV）和单纯疱疹病毒（HSV）活性。