败酱草多糖体外抗呼吸道合胞病毒作用的研究

目的: 了解败酱草多糖(AP4)对呼吸道合胞病毒的体外抑制作用.方法: 经提取、沉淀、离心、大孔吸附树脂两次层析后,得败酱草抗病毒多糖AP4.通过细胞培养法检测呼吸道合胞病毒对Hela细胞的致病变作用及AP4对呼吸道合胞病毒感染Hela细胞的治疗作用.结果: AP4半数中毒浓度(TC50)为11.07mg/ml,抑制呼吸道合胞病毒的半数有效浓度(EC50)为0.097mg/ml,治疗指数(TI)为114;病毒唑半数中毒浓度(TC50)2.087 mg/ml,抑制呼吸道合胞病毒的半数有效浓度(EC50)为0.0385 mg/ml,治疗指数(TI)为54.结论: 败酱草抗病毒有效部位AP4在Hela细胞中对呼吸道合胞病毒有明显的抑制作用.     

抗病毒1号中药有效部位体外对乙型流感病毒的抑制作用

为开发研制安全有效的抗流感病毒新药,本研究采用细胞培养技术,以病毒唑为阳性对照药,观察抗病毒1号中药有效部位(ADl)在MDCK细胞中对乙型流感病毒的抑制作用。结果表明ADl半数中毒浓度(TC_(50))为19.8mg/ml;抗乙型流感病毒的半数有效浓度(EC_(50))为1.47mg/ml;治疗指数(TI)为13.5;染毒后0-8小时分别给药均能抑制病毒增值,且对乙型流感病毒的抑制作用存在明显的量效反应关系。     

硫酸卡那霉素体外抑制呼吸道合胞病毒作用的研究

目的评价硫酸卡那霉素体外抗呼吸道合胞病毒作用机制。方法采用细胞培养技术观察硫酸卡那霉素对细胞的毒性和抗呼吸道合胞病毒作用。结果用空斑减数实验测得半数有效剂量(EC50)为(1.71±0.23)mg/ml,MTT法测定其半数中毒浓度(CC50)为(12.36±0.85)mg/ml,SI为7.2,SI>4(有意义);在不同时间给药对病毒的抑制实验中,呼吸道合胞病毒感染喉癌上皮细胞(Hep-2)细胞后1、2、4、6、8和10h给药均有抑制作用(P<0.05),穿入和吸附抑制实验表明其对病毒穿入过程有明显地抑制作用(P<0.05),但是对吸附过程没有抑制作用。硫酸卡那霉素对RSV病毒没有直接的灭活作用。结论硫酸卡那霉素在体外实验中对呼吸道合胞病毒穿入细胞及在细胞内复制的过程均有抑制作用。     

小儿宝泰康颗粒对呼吸道病毒感染的抗病毒作用研究

目的 观察小儿宝泰康颗粒对呼吸道合胞病毒和腺病毒感染细胞的抑制作用,以便为呼吸道病毒感染的治疗提供新的药物。方法 通过细胞培养技术观察不同浓度的小儿宝泰康颗粒对HEp-2细胞的细胞毒作用,采用MTT法并结合CPE法检测病毒感染细胞存活率和病毒抑制率,以利巴韦林作为阳性药物对照,评价该药物对呼吸道合胞病毒和腺病毒感染细胞的抗病毒活性。结果 小儿宝泰康颗粒对HEp-2细胞的半数细胞毒性浓度（TC₅₀）为4.4 mg·mL^-1,对呼吸道合胞病毒的半数有效浓度（EC₅₀）为2.03 mg·mL^-1,而腺病毒的EC₅₀为0.65 mg·mL^-1,抗病毒指数分别为2.17和6.75。在对两种病毒的直接灭活作用中,EC₅₀分别为4.13和8.99 mg·mL^-1。阳性对照药物病毒唑在128 g·mL^-1时,对呼吸道合胞病毒和腺病毒的抑制率分别为92.7%和80.16%。结论 小儿宝泰康颗粒是一种对呼吸道合胞病毒和腺病毒感染有抗病毒作用的药物,有临床应用前景和进一步研究的价值。     

痰热清抗呼吸道合胞病毒作用体外实验研究

目的研究中草药注射液痰热清对呼吸道合胞病毒(RSV、Long株)的抑制作用。方法以病毒唑为阳性对照药,采用细胞培养技术,观察不同药物浓度及不同给药方式下RSV攻击后各组Hep-2细胞的病变效应(CPE),在此基础上采用MTT比色法,测定各组细胞的病毒抑制率。以CPE法计算药物的半数中毒浓度TC50,分别以CPE法及MTT法计算药物的半数有效浓度EC50及治疗指数TI,比较不同药物浓度及不同给药方式下热毒宁对呼吸道合胞病毒(RSV)的抑制作用效果。结果痰热清注射液半数中毒浓度TC50为3.972g/L,预防给药方式、细胞外直接灭活及治疗给药方式均有抗RSV作用,其抗RSV的半数有效浓度(EC50)分别为0.428/1.160/1.189g/L,治疗指数TI分别是9.28、3.42和3.34,痰热清对RSV的抑制作用存在着明显的量效反应关系。结论痰热清对RSV有直接灭活作用,对RSV侵入Hep-2细胞有阻断作用,对RSV在Hep-2细胞内增殖有抑制作用,相同浓度下,以预防作用更显著。     

单磷酸阿糖腺苷 病毒唑 柴胡及其联合应用对呼吸道合胞病毒的抑制作用

目的研究单磷酸阿糖腺苷(Ara-AMP)、病毒唑、柴胡及其联合应用对呼吸道合胞病毒(RSV)的抑制作用。方法将Ara-AMP、病毒唑及柴胡分别稀释成不同的浓度,用细胞培养法观察药物的细胞毒性反应;采用Vero细胞先感染病毒2h后给药法观察细胞病变(CPE)和药物对RSV的抑制作用。结果Ara-AMP、病毒唑及柴胡对Vero细胞的毒性剂量(TD)分别为0.25、0.2、2mg/ml;当Ara-AMP、病毒唑分别与柴胡联用时并未增加对细胞的毒性,而Ara-AMP与病毒唑联用时则对细胞的毒性增加;3种药物各自在Vero细胞上对RSV都有抑制作用,Ara-AMP和病毒唑的有效剂量均为25滋g/ml,柴胡为125滋g/ml。当3种药物配伍联合应用时,均可产生协同作用。结论Ara-AMP、病毒唑及柴胡各自对RSV具有显著的抑制作用,它们的联合应用可产生显著协同作用,既可降低用药量,减少毒副反应,又可有效抑制RSV的感染。     

菌毒清颗粒体外抗病毒的药效学研究

目的:研究菌毒清颗粒体外抗病毒的药效学。方法:将柯萨奇病毒B族3型(Cox B3)Nancy株、呼吸道合胞病毒(RSV)分别接种于人喉癌上皮Hep2细胞以复制细胞病毒模型。菌毒清颗粒原液(2 mg/ml)与氯原酸原液(2 mg/ml)以1∶2~1∶256倍比稀释后作用于病毒模型细胞,以Reed-Muench法计算50%细胞毒性浓度(TC50)和最大无毒浓度(TC0)。以1.25、0.625、0.312 5、0.15625 mg/ml菌毒清颗粒溶液与0.5、0.25、0.125、0.062 5 mg/ml绿原酸溶液培养病毒模型细胞,显微镜下评价细胞病变程度。以1.25、0.625、0.312 5、0.156 25、0.078 125 mg/ml菌毒清颗粒溶液与0.5、0.25、0.125、0.062 5、0.031 25 mg/ml绿原酸溶液作用于病毒模型细胞,Reed-Muench法计算对RSV、Cox B3细胞的50%抑制浓度(IC50)和治疗指数(TI)。结果:菌毒清颗粒TC50、TC0分别为1.79、1.25mg/ml,绿原酸TC50、TC0分别为0.71、0.5 mg/ml。当菌毒清颗粒溶液质量浓度为1.25 mg/ml时病毒感染细胞生长正常。菌毒清颗粒IC50均为0.22 mg/ml,TI均为8.14;绿原酸IC50分别为0.18、0.36 mg/ml,TI分别为3.94、1.97。结论:菌毒清颗粒具有体外抗Cox B3和RSV病毒的作用。     

小儿双金清热口服液体外抗病毒作用研究

目的观察小儿双金清热口服液体内外的抗病毒作用。方法设置双金药物组、双黄连对照组、利巴韦林对照组、病毒对照组及空白对照组,采用人喉癌细胞培养法检测双金体外抗呼吸道合胞病毒、肠道病毒、柯萨奇B3型病毒、口唇疱疹病毒1型;犬肾细胞培养法检测双金抗甲型流感病毒H1N1病毒作用,采用MTT法检测细胞OD值并计算半数有效浓度、半数中毒浓度及治疗指数。结果体外试验表明双金可抑制呼吸道合胞病毒、肠道病毒、柯萨奇B3型病毒、口唇疱疹病毒1型、甲型流感H1N1病毒的繁殖,半数有效浓度分别为0.003 8、0.006 1、0.003 6、0.008 6、0.001 0 mg·m L-1,半数中毒浓度为对人喉癌细胞半数中毒浓度为0.064 mg·m L-1,对犬肾细胞半数中毒浓度为0.032 mg·m L-1,治疗指数TI分别为16.7、10.4、17.8、32.3。对口唇疱疹病毒1型病毒的繁殖没有抑制作用,TI=7.4;体外试验表明,与双黄连比较,双金的大、中剂量组具有相同的抗甲型流感病毒H1N1、呼吸道合胞病毒的作用。结论双金清热口服液有较好的体外抗呼吸道合胞病毒、肠道病毒、柯萨奇B3型病毒、口唇疱疹病毒1型、甲型流感H1N1病毒作用。     

重组人干扰素γ体外抗呼吸道合胞病毒的作用观察

目的:观察国产重组人干扰素γ(rHuIFN-γ)体外抗呼吸道合胞病毒(respiratory syncytial virus,RSV)的作用。方法:RSV在96孔板中通过Hela细胞进行增殖,不同浓度的IFN-γ加入孔板中,测定药物对RSV感染所致细胞病变的抑制作用。结果:IFN-γ在HeLa细胞上的半数中毒浓度(TC_(50))为2．80万IU·mL~(-1),在HeLa细胞中抗RSV的半数有效浓度(EC_(50))为4．78 IU·mL~(-1),治疗指数(TI)为5857．74。结论:IFN-γ在体外对RSV有显著抑制作用,并呈明显的量效关系。     

病毒唑体外抗副流感病毒(Ⅲ型)作用的实验研究

目的:观察病毒唑体外对副流感病毒( 型)的抑制作用。方法:采用细胞病变抑制实验,观察病毒唑在Hela细胞中对副流感病毒( 型)的抑制作用。结果:病毒唑半数中毒浓度(TC50 )为1783.86μg/ m l;抗副流感病毒( 型)的半数有效浓度(EC50 )为11.5 3μg/ ml,治疗指数(TI)为15 4 .71;病毒唑对副流感病毒( 型)的抑制作用存在明显的量效反应关系(P<0 .0 1) ;在感染病毒后0～2 4 h,每隔2小时加一次药,病毒唑的5 0、2 5、12 .5、6 .2 5 μg/ ml组对副流感病毒( 型)均有抑制作用(P<0 .0 1) ;病毒唑对副流感病毒( 型)有预防和中和作用(P <0 .0 1)。结论:病毒唑在Hela细胞中对副流感病毒( 型)有明显的抑制作用,其作用机制是多途径的。     

热毒宁对流感病毒FM_1的抑制作用

目的:研究中草药热毒宁注射液对流感病毒FM_1的抑制作用。方法:以利巴韦林为阳性对照药,采用细胞培养技术及四氮唑蓝(MTT)比色法,观察在热毒宁干预下,受FM_1感染的犬肾传代细胞(MDCK细胞)的病变效应(CPE)及光密度值(D),计算各组细胞的病毒抑制率。比较不同药物剂量及3种不同给药方式(预防给药、直接灭活及治疗给药)下热毒宁对流感病毒FM_1的抑制作用。结果:热毒宁注射液半数中毒浓度(TC_(50))为10.11g·L~(-1)。预防给药、直接灭活及治疗给药方式下,CPE法测得其抗流感病毒FM_1的半数有效浓度(EC_(50))分别为0.625、0.725、0.629 g·L~(-1),治疗指数(TI)分别为16.17、13.94、16.07;MTT比色法测得其抗流感病毒FM_1的EC_(50)分别为1.135、0.770、0.661 g·L~(-1),TI分别为8.91、13.12和15.29。3种方式给药,MTT法测定时,热毒宁对流感病毒FM_1的抑制作用无差异;CPE法测定时,热毒宁浓度在0.812 51 g·L~(-1)及以上对流感病毒FM_1的抑制作用无差异。结论:热毒宁对流感病毒FM_1有直接灭活及抑制增殖作用,对其感染靶细胞有阻断作用,且呈明显量效关系。     

黄芪多种成分抗人疱疹病毒的初步实验研究

目的：为了开发利用黄芪的抗病毒性质，我们进行了此项研究。方法：我们以阿普洛韦（ACV)为阳性对照，采用对病毒所致细胞病变的抑制及空斑减数实验，观察了黄芪总皂苷、总多糖、总黄酮抗HSV药效。结果：在Hep-2细胞系统中，ACV对HSV－1 HS－1株直接杀灭、感染阻断、增殖抑制的ED50为30．83ug/ml,16.04ug/ml,20.04ug/ml；对HSV－2333株的相应ED50为16.45ug/ml,18.62ug/ml,10.85ug/ml,黄芪总皂苷对HSV－1 HS－1株相应ED50为1．68ug/ml，1．72ug/ml，1．95ug/ml；对HSV－2333株相应ED50为1．73ug/ml，2．70ug/ml，2．74ug/ml，黄芪总多糖对HSV－1 HS－1株的相应ED50为4．03ug/ml，5．33ug/ml，4．90ug/ml；对HSV－2333株相应ED50为6．04ug/ml，5．43ug/ml，7．50ug/ml，黄芪总黄酮对HSV－1 HS－1株的相应ED50为4．95ug/ml，2．75ug/ml，3．49ug/ml；对HSV－2333株的相应ED50为3．56ug/ml，3．93ug/ml，6．27ug/ml。结论：黄芪对HSV－1的治疗指数为ACV的10．3，4．1，5．5倍；对HSV－2的治疗指数为ACV的3．7，2．7，1．0，1．6倍。总多糖对HSV－1的治疗指数为ACV的10．3，4．1，5．5倍；对HSV－2的治疗指数为ACV的3．7，2．7，3．3倍。总黄酮的治疗指数为ACV相近，值得开发利用。     

异莲心碱的体外抗氧化活性

目的研究异莲心碱对氧自由基的清除作用及其对脂质过氧化反应(LPO)的抑制作用。方法羟自由基由Fenton体系产生,超氧阴离子自由基由邻苯三酚自氧化法产生,通过比色法测定LPO的终产物 丙二醛(MDA)的相对含量来反映异莲心碱对LPO的影响。结果异莲心碱对羟自由基和超氧阴离子自由基有较强的清除作用,达到5 0 %清除率所需药物浓度(EC50 )分别为0 .90 ,1.82mg/ml,可抑制小鼠肝匀浆脂质过氧化反应,EC50 为0 .6 7mg/ml。结论异莲心碱对氧自由基有清除作用,对脂质过氧化有抑制作用,其活性与剂量呈正相关     

体外筛选具有抗柯萨奇病毒作用的中药提取物

目的通过建立体外抗病毒药物筛选平台,寻找具有抗柯萨奇病毒的中药。方法根据文献报道,候选39种可能具有抗病毒作用的中药,对其有效部位进行水体物和醇提物,共获得160种中药有效部位。培养Hela细胞,接种病毒和药物,通过以噻唑蓝(MTT)染色方法为主的细胞病变抑制实验和病毒繁殖抑制实验对上述药物进行筛选,之后通过公式计算有明确抗病毒作用药物的半数有效剂量(IC50)和半数致死量(CC50)。结果在候选药物的有效部位中,黄芪甲苷和红景天苷具有显著抗病毒作用,细胞病变(CPE)抑制实验结果显示黄芪甲苷的半数有效量(IC50)为(6.4±0.5)μg/ml、红景天苷IC50值为:(9.6±1.4)μg/ml;黄芪甲苷半数毒性剂量(CC50)为(1287±182)μg/ml、红景天苷CC50值为:(952±83)μg/ml。2种提取物的的治疗指数(SI)分别为:200和99。结论黄芪水体物和醇提物、红景天醇提物具有抗病毒作用,其细胞毒性均低于对照药利巴韦林,其中黄芪醇提物的体外抗病毒效果与利巴韦林接近。     

Prophylaxis of Rift Valley fever with antiviral drugs, immune serum, an interferon inducer, and a macrophage activator

Rift Valley fever virus (RVFV), a member of the family Bunyaviridae, extended its range from sub-Saharan Africa into Egypt in 1977. Its clinical spectrum is recognized to include severe manifestations such as hemorrhagic fever and encephalitis. For these reasons, as well as the limited knowledge of specific therapy for Bunyaviridae infections, we investigated several prophylactic regimens for RVF in a mouse model. Rimantadine, thiosemicarbazone, and inosiplex were ineffective. Pretreatment with glucan was of some use, but the most encouraging results were obtained with the antiviral drug ribavirin, passive antibody, or an interferon inducer polyriboinosinic-polyribocytidylic acid complexed with poly-L-lysine and carboxymethylcellulose (poly[ICLC]). Ribavirin and poly(ICLC) were also shown to be efficacious in preventing disease in hamsters. Ribavirin (loading dose of 50 mg/kg followed by 10 mg/kg at 8-h intervals for 9 days) suppressed viremia in RVF-infected rhesus monkeys. Ribavirin also reduced virus yield in infected cell cultures; sensitivity varied markedly with cell type but not with virus strain. Immune mouse ascitic fluid, with a plaque reduction neutralization titer of 1:1024, was effective in a dose of 4 ml/kg, a volume approximately equivalent to administration of a unit of convalescent plasma to a human. Poly(ICLC) may well have functioned through interferon induction, since RVFV was shown to be sensitive to interferon in cell culture, and since another macrophage activator (glucan) was only marginally effective. These studies suggest that ribavirin, poly(ICLC), and convalescent plasma may have a role in prevention or therapy of human RVF.     

Chromosomal instability and cytotoxicity induced by ribavirin: comparative analysis in cell lines with different drug-metabolizing profiles

Ribavirin is an important component of the treatment for hepatitis C virus (HCV) infection and, in combination with the new direct-acting antiviral (DAA) agents, comprises the major current therapeutic regimens. This study evaluated the cytotoxicity and chromosomal instability induced by ribavirin using the in vitro cytokinesis-block micronucleus cytome (CBMN-Cyt) assay in two cell lines with different expression levels of drug-metabolizing enzymes: human hepatocellular carcinoma cells (HepG2) and Chinese hamster ovary (CHO-K1) cells. HepG2 cells were treated with nine concentrations (from 15.3?μg/ml to 3.9?mg/ml) and CHO-K1 cells were exposed to eight concentrations (from 15.3?μg/ml to 1.9?mg/ml) of ribavirin for 24?h. Ribavirin inhibited cell proliferation in both cell lines, but at different concentrations: 3.9?mg/ml in HepG2 and 244.2?μg/ml in CHO-K1 cells. No significant differences were observed regarding aspects of cell death in HepG2 and CHO-K1 cells, reflecting the absence of cytotoxic effects associated to ribavirin. Ribavirin did not increase the frequency of nucleoplasmic bridges (NPBs) and nuclear bud (NBUD). However, when compared to the negative control, a significant increase in micronuclei (MNi) frequency was observed in both cell lines. However, chromosomal instability was induced by higher concentrations of ribavirin in HepG2 cells (from 61.1 to 976.8?μg/ml), compared with CHO-K1 cells (15.3 and 30.5?μg/ml). These results demonstrate the potential of ribavirin to promote chromosomal instability, and suggest that cells with different expressions of drug-metabolizing enzymes show different susceptibility to ribavirin effects.     

Inhibition of hepatitis delta virus RNA replication in primary woodchuck hepatocytes

Cell cultures of primary woodchuck hepatocytes can be infected with hepatitis delta virus (HDV) as demonstrated by the appearance of genomic HDV RNA 7 days after inoculation. This tissue culture system was used to study the effect of antiviral substances. Ribavirin inhibited HDV replication at a concentration of 10 micrograms/ml, if added up to three days post infection. Suramin had an inhibitory effect only when added simultaneously with the virus, at a concentration of 200 micrograms/ml. This concentration had no toxic effect on primary woodchuck hepatocytes. alpha-Amanitin showed a weak inhibitory effect only at the highest nontoxic concentration of 0.1 microgram/ml. Acyclovir had no effect.     

Antiviral activity of hop constituents against a series of DNA and RNA viruses

We investigated whether crude hop extracts and purified hop components representing every major chemical class of hop compound have antiviral activity. These hop constituents were tested for antiviral activity against bovine viral diarrhea virus (BVDV) as a surrogate model of hepatitis C virus (HCV), human immunodeficiency virus (HIV), influenza A virus (FLU-A), influenza B virus (FLU-B), rhinovirus (Rhino), respiratory syncytial virus (RSV), yellow fever virus (YFV), cytomegalovirus (CMV), hepatitis B virus (HBV), and herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2). The extracts all failed to prevent the replication of HIV, FLU-A, FLU-B, RSV and YFV. A xanthohumol-enriched hop extract displayed a weak to moderate antiviral activity against BVDV (therapeutic index (TI)=6.0), HSV-2 (TI=>5.3), Rhino (TI=4.0) and HSV-1 (TI=>1.9) with IC(50) values in the low microg/ml range. Pure iso-alpha-acids demonstrated low to moderate antiviral activity against both BVDV (TI=9.1) and CMV (TI=4.2) with IC(50) values in the low microg/ml range. No antiviral activity was detected using beta-acids or a hop oil extract. Ultra-pure preparations (>99% pure) were used to show that xanthohumol accounted for the antiviral activity observed in the xanthohumol-enriched hop extract against BVDV, HSV-1 and HSV-2. Xanthohumol was found to be a more potent antiviral agent against these viruses than the isomer iso-xanthohumol. With Rhino, the opposite trend was observed with iso-xanthohumol showing superior antiviral activity to that observed with xanthohumol. Xanthohumol also showed antiviral activity against CMV, suggesting that it might have a generalized anti-herpesvirus antiviral activity. Again, superior antiviral activity was observed with the xanthohumol isomer against CMV. In summary, iso-alpha-acids and xanthohumol were shown to have a low-to-moderate antiviral activity against several viruses. These hop constituents might serve as interesting lead compounds from which more active anti-HCV, anti-Rhino and anti-herpesvirus antiviral agents could be synthesized.