病毒灵喷雾剂抗单纯疱疹病毒的实验研究

目的:研究病毒灵喷雾剂对单纯疱疹病毒(HSV)的作用。方法:采用细胞培养体外测定病毒灵喷雾剂对HSV-1或HSV-2感染Hep-2细胞的半数抑制浓度EC50和治疗指数(TI)。在豚鼠上观察病毒灵喷雾剂对HSV-1感染皮肤的治疗作用,在小鼠上观察对HSV-2感染阴道炎的治疗作用。结果:体外实验表明,病毒灵喷雾剂对HSV-1、HSV-2致细胞病变均有不同程度的抑制作用;体内实验表明,病毒灵喷雾剂对HSV-1感染豚鼠皮肤及HSV-2感染小鼠阴道所致病变均有不同程度的抑制作用。结论:病毒灵喷雾剂具有抗HSV的作用。     

灵芝中酸性蛋白结合多糖单用或与阿昔洛韦和阿糖腺苷合用的抗疱疹病毒活性

为寻找抗疱疹病毒活性成分,作者以生物活性为指导,从灵芝子实体水溶性物质中分得一棕色酸性蛋白结合多糖(APBP)。抗单纯疱疹病毒Ⅰ型(HSV-1)和Ⅱ型(HSV 2)的实验显示,APBP对Vero细胞中的HSV1和HSV-2均具强抗病毒活性,EC_(50)分别为     

广西苦丁茶提取物体外抗单纯疱疹病毒1型活性的实验研究

目的探讨不同苦丁茶提取物体外抗单纯疱疹病毒1型（HSV-1）的作用。方法采用细胞病变效应法（Cytopathogenic effect，CPE）和空斑减数实验（Plaque reduction assay）测定不同苦丁茶提取物抗HSV-1活性，计算药物对病变的抑制率和半数抑制浓度（50％ inhibiting concentration，IC50），并从药物对细胞的保护，对病毒增殖的影响及对病毒感染细胞的综合作用3个方面初步探索苦丁茶提取物抗HSV-1病毒活性的机理。结果苦丁茶水提物能明显抑制HSV-1的致病变作用，IC50为108．24μg／ml，药物对细胞的保护及对病毒感染细胞的综合作用中均表现出明显的活性。结论苦丁茶水提物具有显著的抗HSV-1活性，且初步推测其抗病毒活性是作用在病毒和受体结合，侵入Vero细胞阶段。     

夏枯草酶解水提液分级醇沉及其抗HSV-1活性

目的探究夏枯草酶解水提液分级醇沉部分的成分组成及其抗HSV-1活性。方法利用纤维素酶酶解法选择性提高夏枯草多糖的提取率,对其酶解水提液进行分级醇沉,用细胞病变效应(CPE)实验对各浓度的乙醇沉淀进行活性测试,根据细胞试验计算EC_(50)。筛选出夏枯草水提液中抗HSV活性最优的部分,采用大孔树脂D301对夏枯草酶解水提液分级醇沉中具有抗HSV-1活性的部分进行分离纯化,除去多酚及蛋白质类成分,得到多糖含量较高的部分,并测试该部分的抗HSV活性。结果夏枯草粉末经纤维素酶酶解后的水提液分级醇沉沉淀中,20%、30%乙醇的沉淀具有良好的抗HSV-1活性,其ED_(50)分别为60.79、42.45μg/mL,40%乙醇以上的分级沉淀无活性或活性较弱。经大孔树脂D301纯化后的夏枯草多糖纯度可达77.78%,但其抗HSV活性反而丧失。结论夏枯草多糖含量高低与其抗HSV活性强弱并不呈正相关,可为进一步研究研究其有效部位及构效关系提供参考。     

青羊参多糖硫酸酯体外抗单纯疱疹病毒Ⅱ型活性研究

目的 体外测定青羊参多糖硫酸酯抗单纯疱疹病毒Ⅱ型的活性。方法采用MTT法检测化合物对疱疹病毒感染Vero细胞的保护作用。结果青羊参多糖硫酸酯体外抗HSV—Ⅱ的EC50为69．9μg／ml，治疗指数为103。结论在体外，青羊参多糖硫酸酯有较显著的抗HSV-Ⅱ的作用。     

马齿苋提取物体外抗Ⅱ型单纯疱疹病毒的活性评价

目的利用体外细胞培养技术初步筛选马齿苋抗Ⅱ型单纯疱疹病毒（HSV-2）的有效部位。方法以选择性指数（Selectivity-Index,SI）为活性评价指标,通过细胞毒实验及抗病毒活性评价实验,对马齿苋的不同提取部位进行了体外抗HSV-2病毒活性的筛选研究。结果马齿苋50%乙醇提取部位显示了潜在的抗HSV-2病毒活性,IC50值为120μg/mL,SI值为15;水提取物以醋酸纤维素膜分离后的粗多糖部位则显示了较强的抗HSV-2病毒活性,IC50值为55μg/mL,SI值为34。结论马齿苋抗HSV-2病毒活性成分为多糖类成分。     

023 杉藻属植物Gigartina skottsbergii角叉菜聚糖抗小鼠腹腔单纯疱疹病毒的作用

先前研究表明，从杉藻属红藻类植物G．skottsber gii中分离的多种结构类型的天然角叉菜聚糖在体外实验中对单纯疱疹病毒（HSV）-1和-2复制具有选择性抑制活性，其中环化μ／v-角叉菜聚糖1C3活性较强而无细胞毒性。作者采用体内实验，首次研究了1C3对鼠腹膜内HSV-1感染的抗病毒活性。     

金线吊乌龟中生物碱的抗单纯疱疹病毒作用的研究

近10年来用于抗单纯疱疹病毒(HSV-1和HSV-2)的无环鸟苷及其它核苷衍生物已经出现了抗药性。本次旨在寻找新的抗HSV的药物,尤其对中药金线吊乌龟(Stephania cepharantha)中49种生物碱的抗HSV活性进行了研究。 实验材料:采用猴肾脏细胞。HSV-1、TK-HSV-1和HSV-2病毒均冷冻保存。     

夏枯草水提物体外抗单纯疱疹病毒1型作用的研究

目的:研究夏枯草水提物体外抗单纯疱疹病毒1型(HSV-1)作用。方法:以Vero细胞作为病毒感染靶细胞,给予不同浓度的药物处理,镜下观察细胞的形态变化,应用MTT法检测夏枯草水提物体外抗HSV-1的效果。并以不同浓度夏枯草水提物作用于HSV-1感染的不同阶段,探讨夏枯草水提物抗HSV-1的机制。结果:夏枯草水提物对HSV-1具有明显的抑制作用,随着浓度的增加,抗病毒作用增强,呈量效关系,TI(药物治疗指数)值为45.71。进一步实验发现夏枯草水提物6.25 mg生药/ml在体外即能抑制病毒吸附和生物合成,又能直接杀灭病毒。结论:夏枯草水提物具有显著的抗HSV-1作用,是对病毒感染不同阶段综合作用的结果。     

夏枯草抗疱疹病毒多糖的提取及纯化工艺研究

目的:优化夏枯草抗疱疹病毒多糖的提取及纯化工艺,为新药研究提供科学依据。方法:以多糖含量和浸膏得率为指标,采用L9(34)正交试验法对夏枯草多糖提取过程中的提取次数,加水倍量及提取时间3个因素进行优化研究;以多糖含量及体外抗单纯性疱疹病毒I型(HSV-1)活性为考察指标,对超滤和醇沉两种纯化技术进行比较。结果:最优水提工艺为第1次加水14倍,第2、3次加水12倍,提取3次,每次提取时间1.5 h;采用超滤和醇沉两种技术分别纯化所得多糖。通过MTT法测得醇沉多糖和超滤多糖抑制HSV-1的IC50分别为(93.99±13.12)μg/m L和(51.35±15.30)μg/m L,结果表明超滤技术制备的多糖提取物抗单纯性疱疹病毒活性显著高于醇沉技术。结论:采用超滤技术制备的夏枯草多糖具有较高的含量及抗单纯性疱疹病毒活性,可作为夏枯草多糖开发新药的纯化工艺。     

In vitro and in vivo activity of eugenol on human herpesvirus

Eugenol (4-allyl-1-hydroxy-2-methoxybenzene) was tested for antiviral activity against HSV-1 and HSV-2 viruses. In vitro, it was found that the replication of these viruses was inhibited in the presence of this compound. Inhibitory concentration 50% values for the anti-HSV effects of eugenol were 25.6 microg/mL and 16.2 microg/mL for HSV-1 and HSV-2 respectively, 250 microg/mL being the maximum dose at which cytotoxicity was tested. Eugenol was virucidal and showed no cytotoxicity at the concentrations tested. Eugenol-acyclovir combinations synergistically inhibited herpesvirus replication in vitro. Topical application of eugenol delayed the development of herpesvirus induced keratitis in the mouse model.     

Anti-viral activity of the extracts of a Kenyan medicinal plant Carissa edulis against herpes simplex virus

Herpes simplex virus (HSV) infection is a major opportunistic infection in immunosuppressed persons. It is therefore a serious disease in high HIV/AIDS prevalence areas as in sub-Saharan Africa where infections due to HSV have risen significantly. The development of resistant strains of HSV to the available drugs for infection management, as is evident in the first drug of choice acyclovir, has further compounded this situation. There is therefore an urgent need to identify and develop new alternative agents for management of HSV infections, more so, for those due to resistant strains. We report here on an aqueous total extract preparation from the roots of Carissa edulis (Forssk.) Vahl (Apocynaceae), a medicinal plant locally growing in Kenya that has exhibited remarkable anti-HSV activity in vitro and in vivo for both wild type and resistant strains of HSV. The extract significantly inhibited formation of plaques in Vero E6 cells infected with 100PFU of wild type strains of HSV (7401H HSV-1 and Ito-1262 HSV-2) or resistant strains of HSV (TK(-) 7401H HSV-1 and AP(r) 7401H HSV-1) by 100% at 50 microg/ml in vitro with minimal cell cytotoxicity (CC(50)=480 microg/ml). When the extract was examined for in vivo efficacy in a murine model using Balb/C mice cutaneously infected with wild type or resistant strains of HSV, the extract at an oral dose of 250 mg/kg significantly delayed the onset of HSV infections by over 50%. It also increased the mean survival time of treated infected mice by between 28 and 35% relative to the infected untreated mice (p<0.05 versus control by Student's t-test). The mortality rate for mice treated with extract was also significantly reduced by between 70 and 90% as compared with the infected untreated mice that exhibited 100% mortality. No acute toxicity was observed in mice at the oral therapeutic dose of 250 mg/kg. These results suggest that this herbal extract has potent anti-viral agents against herpes simplex viruses that can be exploited for development of an alternative remedy for HSV infections.     

大豆总甙对病毒的抑制作用及其临床应用的研究

大豆总甙（TotalSoyasaponinTS）对单纯疱疹病毒Ⅰ型（HSV－I）、柯萨奇Bs（CoxBs）病毒的复制有明显的抑制作用；同时体现出对病毒感染细胞很强的保护作用。其作用机理与TS对病毒的直接杀伤作用有密切关系。临床应用研究表明，对疱疹性口唇炎和口腔溃疡效果显著，有效率达88．8％和76．9％。本研究为应用TS治疗病毒性疾病提供了一定的实验依据。     

Anti-herpes simplex virus activities of Eugenia caryophyllus (Spreng.) Bullock & S. G. Harrison and essential oil, eugenol

In this study, an extract from the flower buds of Eugenia caryophyllus (Spreng.) Bullock & S. G. Harrison and the essential oil, eugenol, were evaluated for their anti-herpes simplex virus properties on standard HSV-1(F), standard HSV-2(G) and ten HSV isolates. The plaque reduction assay showed that HSV-1(F), HSV-2(G), two HSV-1 isolates (2, 30) and four HSV-2 isolates (1, 2, 3, 21) were inhibited by E. caryophyllus. Only HSV-1 isolates 1 and 30 were inhibited by eugenol. Thus, strains or isolates of viruses may affect the range of inhibition. Moreover, particles of HSV standard strains were directly inactivated by E. caryophyllus and eugenol. The total virus yield of HSV standard strains and isolates at 30 h also declined after treatment with E. caryophyllus and eugenol. The E. caryophyllus extract exerted higher antiviral replication on HSV-2(G) than on HSV-1(F). The inhibition of the viral yield of HSV-1 isolates was higher than standard HSV-1(F) and standard HSV-2(G) was also inhibited more than most of the HSV-2 isolates. The anti-HSV activity of eugenol against HSV-1(F) and HSV isolates was stronger than with the E. caryophyllus crude extract. However, the percentage inhibition was more pronounced on HSV-1(F) than on HSV-2(G). Moreover, HSV-1(1) and HSV-2(1, 32) could not replicate when eugenol was included in the assay.     

Possible mode of antiviral activity of acidic protein bound polysaccharide isolated from Ganoderma lucidum on herpes simplex viruses

Two protein bound polysaccharides, a neutral protein bound polysaccharide (NPBP) and an acidic protein bound polysaccharide (APBP), were isolated from water soluble substances of Ganoderma lucidum by EtOH precipitation and DEAE-cellulose column chromatography. Their antiviral activities against herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) were then investigated by plaque reduction assay. APBP exhibited more potent HSV-1 and HSV-2 antiviral activity than NPBP with 50% effective concentration (EC(50)) of 300-520 microg/ml. In order to examine the possible mode of the antiviral activity of APBP its virucidal effect, antiviral activity in preincubation, attachment and penetration assay were tested with HSV-1 and HSV-2. APBP was found to have a direct virucidal effect on HSV-1 and HSV-2. APBP did not induce IFN or IFN-like materials in vitro and is not expected to induce a change from a normal state to an antiviral state. APBP in concentrations of 100 and 90 microg/ml inhibited up to 50% of the attachment of HSV-1 and HSV-2 to Vero cells and was also found to prevent penetration of both types of HSV into Vero cells. These results show that the antiherpetic activity of APBP seems to be related to its binding with HSV-specific glycoproteins responsible for the attachment and penetration, and APBP impedes the complex interactions of viruses with cell plasma membranes.     

In vivo antiviral activity of Stephania cepharantha against herpes simplex virus type-1

The antiviral activity of a MeOH extract of Stephania cepharantha (root tubers), its CHCl3-soluble fraction (alkaloid fraction) and the major alkaloid FK-3000 (1) were investigated in BALB/c mice cutaneously infected with HSV-1 strain 7401H. At doses of 125 and 250 mg/kg body weight, p.o., the MeOH extract significantly delayed skin lesion on score 2 (vesicles in the local region), limited the development of further lesions on score 6 (mild zosteriform lesion) and prolonged the mean survival time of HSV-1 infected mice. After p.o. administration of the CHCl3-soluble fraction at doses of 25 and 50 mg/kg or FK-3000 (1) at 10 and 25 mg/kg, similar results were obtained. Although the alkaloid improved the survival of infected mice, it had a narrow therapeutic index.     

滨蒿总黄酮提取物的抗流感病毒作用

目的:观察滨蒿总黄酮提取物(滨蒿总黄酮)在体内外的抗流感病毒作用。方法:体外抗病毒实验观察样品对狗肾传代细胞(MDCK)的毒性,观察病毒致细胞病变(CPE)法测定样品在体外抑制甲、乙型流感病毒的半数抑制浓度(IC50),并计算选择指数(SI)。体内抗病毒实验用流感病毒鼠肺适应株(PR8)滴鼻感染小鼠建立流感病毒性肺炎模型,以死亡保护率、生命延长率和肺指数等为指标,评价样品的保护作用。结果:滨蒿总黄酮对甲、乙型流感病毒致细胞病变的IC50分别为74.6μg/mL、98.5μg/mL,SI分别为16.8、12.7;滨蒿总黄酮以200、400mg/kg剂量灌胃给药对小鼠流感病毒性肺炎模型有明显的保护作用,其死亡保护率分别为36.4%、54.5%;生命延长率分别为29.0%、38.2%;滨蒿总黄酮400mg/kg剂量对流感病毒性肺炎小鼠肺指数有明显的抑制作用,抑制率为25.9%。结论:滨蒿总黄酮在体内外均有明显的抗病毒作用。     

Proanthocyanidin-enriched extract from Myrothamnus flabellifolia Welw. exerts antiviral activity against herpes simplex virus type 1 by inhibition of viral adsorption and penetration

Aim of the study

Extracts from the aerial parts of the South African resurrection plant Myrothamnus flabellifolia Welw. have been used traditionally against infections of the upper respiratory tract and skin diseases. A polyphenol-enriched extract was investigated for potential antiviral effects against herpes simplex virus type 1 (HSV-1) and adenovirus, and the underlying mode of action was to be studied.

Materials and methods

Antiviral effects of an acetone-water extract (MF) from Myrothamnus flabellifolia on HSV-1 and adenovirus type 3 were tested in infected Vero cells by plaque reduction assay, MTT test and immunofluorescence. The influence of the extract on the HSV-1 envelope glycoprotein D was shown by Western blot. Organotypic full thickness skin models consisting of multilayer skin equivalents were used for the investigation of MF effects on HSV-1 replication.

Results

MF exhibited strong antiviral activity against HSV-1. The HSV-1-specific inhibitory concentration (IC₅₀) was determined as 0.4 μg/mL and the cytotoxic concentration (CC₅₀) against Vero cells as 50 μg/mL. A selectivity index (SI) (ratio of CC₅₀ to IC₅₀) of approximately 120 was calculated when MF was added to the virus inoculum for 1 h at 37 °C prior to infection. The replication of adenovirus 3 was not affected by MF.MF abolished virus entry into the host cell by blocking viral attachment to the cell surface. When added after attachment at a concentration of >6 μg/mL, the extract also inhibited penetration of HSV-1 into the host cell. Polyphenolic compounds from MF directly interacted with viral particles, leading to the oligomerisation of envelope proteins as demonstrated for the essential viral glycoprotein D (gD). Using organotypic full thickness tissue cultures, it was shown that treatment of HSV-1 infected cultures with the MF resulted in reduced viral spread.

Conclusions

A polyphenol-enriched extract from Myrothamnus flabellifolia strongly acts against HSV-1 by blocking viral entry into the cells.     

蚬壳花椒果皮多糖抗肿瘤活性研究

目的:研究蚬壳花椒果皮多糖的体内外抗肿瘤活性。方法:采用水提醇沉法制备蚬壳花椒果皮多糖,以MTT法和移植性H22实体荷瘤小鼠为模型,检测不同剂量蚬壳花椒果皮多糖的体内外抗肿瘤作用。结果:当质量浓度为62.5～1 000μg/mL时,其对体外培养的2株人肿瘤细胞Hela和SMMC-7721的增殖表现出显著的抑制作用,IC50依次为460.9μg/mL、425.6μg/mL。体内实验表明,蚬壳花椒果皮多糖高(1.5g/kg)、中(0.75g/kg)、低(0.375g/kg)剂量组均能抑制荷瘤小鼠瘤块的生长,抑瘤率依次为41.55%、34.96%、33.71%;通过肿瘤组织病理学切片观察,同样显示蚬壳花椒果皮多糖具有较好的抗肿瘤作用。比较给药后荷瘤小鼠的肝脏指数、脾脏指数和胸腺指数后发现,蚬壳花椒果皮多糖毒副作用小,增强免疫功能。结论:蚬壳花椒果皮多糖在体内外均有明显的抗肿瘤活性。     

喷昔洛韦对单纯疱疹病毒感染小鼠的治疗作用

 目的研究喷昔洛韦(PCV)对单纯疱疹病毒感染小鼠的疗效。方法用病毒滴度为100 LD₅₀/0.1 mL HSV-1，HSV-2每只分别腹腔注射0.03 mL，24 h后皮下注射给药，PCV剂量分别为20，50，100，250 mg·kg^-1·d^-1,疗程5d。观察动物的反应，出现死亡时间和死亡数，所有动物用药后观察15 d。同时收集各组动物治疗后的标本，进行病毒分离，按常规测定病毒滴度。另设病毒对照组、0.9% NaCl对照组、阳性药阿昔洛韦对照组(ACV)。结果 PCV对HSV感染鼠有保护作用，50 mg·kg^-1·d^-1时，HSV-1和HSV-2感染鼠的存活率分别为60%和75%，平均存活天数分别为(11.3±3.6)和(12.9±2.8)d，随着药物剂量增加，疗效增强，该药还能降低组织内病毒滴度，阻止体内病毒复制，抗HSV作用类似于相同剂量的ACV。结论PCV有抗HSV作用。