飞龙掌血中心血管活性成分的分离与鉴定

从飞龙掌血乙醇提取物的乙醚萃取部分分离到一晶体,药理实验表明该晶体具有心血管药理活性。经理化和波谱分析,鉴定其为异茴芹香豆素。     

飞龙掌血醇提物的抗炎镇痛作用

目的研究飞龙掌血醇提物的抗炎镇痛作用及其机制。方法大鼠用角叉菜胶致炎及福尔马林致痛,研究飞龙掌血醇提物的镇痛作用;ELISA试剂盒检测角叉菜胶实验大鼠血清中β内啡肽(β-EP)、前列腺素E2(PGE2)、5-羟色胺(5-HT)的含有量,炎症大鼠皮肤组织中肿瘤坏死因子-α(TNF-α)及白细胞介素-1β(IL-1β)的含有量,以及福尔马林实验小鼠血清中白三烯B4(LTB4)的含有量;免疫组化法观察大鼠脊髓FOS及P物质(SP)蛋白表达。结果飞龙掌血醇提物能明显降低足趾肿胀率;福尔马林实验中,飞龙掌血醇提物能明显减少舔足累积时间,降低PGE2的含有量,高剂量组尤为明显;3个剂量组均能降低血清中5-HT的含有量,高剂量和低剂量效果显著,高剂量组还能升高血清中β-EP的含有量。各剂量组均能降低皮肤组织中TNF-α和IL-1β的含有量,中、高剂量组抑制FOS蛋白表达;高剂量组明显降低血清中LTB4的含有量。结论飞龙掌血醇提物具有抗炎镇痛作用,其作用机制可能是与通过增加血清中β-EP,降低PGE2、5-HT、LTB4含有量,降低皮肤组织中TNF-α、IL-1β含有量,以及下调脊髓SP及FOS蛋白表达有关。     

飞龙掌血的化学成分、药理活性及质量评价研究进展

通过梳理近年来飞龙掌血的相关文献,对其化学成分、药理活性及毒理作用进行了总结。飞龙掌血现行的质量标准仅对白屈菜红碱进行了含量测定,无法全面评价其品质。不同产地、采收季节、炮制方法对飞龙掌血的有效成分也有影响,通过归纳近年来对飞龙掌血质量评价的相关文献,提出采用多种质量标志物对其质量进行评价,为建立更加全面的飞龙掌血标准提供依据。     

飞龙掌血提取物的镇痛作用及相关机制研究

  目的：对飞龙掌血的镇痛作用及相关机制进行实验探讨,为进一步研究开发镇痛中药提供参考  方法：采用醋酸扭体法和热板法考察飞龙掌血的提取物对于化学刺激和热刺激引起疼痛的镇痛效果;用酶联免疫法(ELISA法)检测小鼠血清中β内啡肽(β-EP)、前列腺素E₂(PGE₂)、五羟色胺(5-HT)、一氧化氮(NO)的含量;用Western blotting法检测小鼠脑组织中β-EP、PGE₂和阿片μ受体的蛋白表达  结果：飞龙掌血提取物能明显减少冰醋酸所致小鼠的扭体次数,且醇提物低于水提物(P<0.01);飞龙掌血提取物能提高小鼠热刺激痛阈值,醇提物与水提物之间无显著性差异(P>0.05);飞龙掌血提取物能增加小鼠血清中的β-EP的含量、降低PGE₂及NO的含量,且醇提物更明显(P<0.05),而5-HT的含量无变化(P>0.05);飞龙掌血的提取物还能上调小鼠脑组织中β-EP的蛋白,下调PGE₂的的蛋白表达(P<0.05)  结论：飞龙掌血具有镇痛作用,且醇提物效果优于水提物,高剂量组优于中、低剂量组,其镇痛机制可能是与通过增加血清中β-EP,降低PGE₂、NO的含量,上调脑组织中β-EP的蛋白表达、下调PGE₂的表达有关。     

飞龙掌血根皮的生药学鉴别与止血活性考察

目的:开展飞龙掌血根皮的组织切片、粉末显微方面的生药学鉴别研究,以及其止血活性与样品制备方法研究。方法:通过制作飞龙掌血全根横切片,飞龙掌血根皮与根心各自粉末的水装片、透化片、染色片,运用生药学传统鉴别手段完善飞龙掌血全根的组织切片特征,寻找飞龙掌血根皮与根心之间的粉末显微特征。采用小鼠断尾法、毛细玻管法,以出血时间、出血量、凝血时间为指标,考察不同提取方法、不同极性部位对飞龙掌血根皮止血活性的影响。结果:飞龙掌血根皮含有大量石细胞、草酸钙方晶、木栓细胞、油细胞等,飞龙掌血根心含木纤维、草酸钙方晶、网纹纹孔导管等。95%乙醇冷浸提取与50%乙醇回流提取都适合于飞龙掌血根皮提取;冷浸浸膏乙酸乙酯部位止血活性优于石油醚部位和正丁醇部位,冷浸浸膏乙酸乙酯部位按剂量1.50 g·kg-1给药,平均出血时间、出血量、凝血时间依次为(59.67±12.31)s,(4.42±1.67)mg,(79.67±5.57)s。结论:石细胞、木栓细胞、油细胞是飞龙掌血根皮的显微鉴别项目,木纤维是飞龙掌血根心的专有鉴别项目。飞龙掌血根皮的提取应结合乙醇冷浸法和回流法,飞龙掌血根皮具有良好的止血凝血效果,以冷浸提取乙酸乙酯部位止血效果最好。     

飞龙掌血乙醇提取物对白色念珠菌抑菌作用的研究

目的:研究飞龙掌血乙醇提取物体外抑制白色念珠菌生长作用,为进一步开发根管消毒新药提供理论依据.方法:微量肉汤稀释法确定飞龙掌血乙醇提取物体外抑制白色念珠菌生长的最小抑菌浓度(MIC)及最小杀菌浓度(MBC);常规扫描电镜制样,观察细菌细胞形态的变化;采用RT-PCR技术获得互补DNA,PCR技术扩增白色念珠菌毒力因子SNF2和PDE2基因片段,产物进行电泳分析.结果:飞龙掌血乙醇提取物对白色念珠菌的最小抑菌浓度为7.5 g·L-1,最小杀菌浓度为15.0 g·L-1;扫描电镜观察药物作用后菌体呈现不规则形态,表面有明显的凹陷、裂痕;随着药物浓度增加,毒力因子SNF2和PDE2的表达量呈明显下降趋势.结论:飞龙掌血乙醇提取物对白色念珠菌有较好的抑菌活性;推断该药物是通过抑制或干扰毒力因子SNF2和PDE2转录过程的表达,从而抑制致病性菌丝形态的形成及破坏细菌细胞壁的完整性,达到抑菌作用.     

两面针的化学成分和飞龙掌血酮内酯的晶体结构

目的 研究两面针Zanthoxylum nitidum的化学成分。方法 应用硅胶柱色谱和Sephadex LH-20凝胶柱色谱分离化合物,通过化合物的理化性质和光谱数据(包括单晶X射线衍射)鉴定结构。结果 从两面针中分离得到7个化合物,分别为2个香豆素：飞龙掌血酮内酯(Ⅰ)和飞龙掌血内酯(Ⅱ);5个生物碱：6-甲氧基-5,6-二羟基两面针碱(Ⅲ)、白鲜碱(Ⅳ)、岩椒碱(Ⅴ)、茵芋碱(Ⅵ)和5-甲氧基白鲜碱(Ⅶ)。对化合物Ⅰ进行了单晶X射线衍射实验。结论 化合物Ⅰ～Ⅲ、Ⅴ、Ⅶ为首次从该植物中分离得到;首次得到化合物Ⅰ的单晶X射线衍射晶体结构数据。     

苍术抗流感病毒有效成分的筛选

目的从苍术中筛选抗流感病毒的有效成分。方法采用水提、超临界二氧化碳流体(SFE-CO2)提取,并利用硅胶柱层析法进行成分分离,得到苍术的各种提取物和有效成分单体,再用流感病毒甲型H3N2、H5N1(禽流感病毒)和乙型流感病毒进行狗肾传代细胞株(MDCK)组织培养筛选。结果含苍术酮的苍术SFE-CO2提取物及苍术酮,对流感病毒有杀灭作用;而苍术素、苍术的水提液对流感病毒无抑制作用。结论苍术酮是苍术中抗流感病毒的有效成分。     

甘草抗病毒有效部位体外抗副流感病毒(Ⅲ型)作用的研究

目的：研究甘草抗病毒有效部位（GC3—1—4）体外抑制副流感病毒（Ⅲ型）的作用。方法：采用煎煮法和柱层析法提取分离得到甘草抗病毒有效部位（GC3—1—4）。以病毒唑为阳性对照药，采用细胞病变抑制实验观察GC3—1—4在Hela细胞中对副流感病毒（Ⅲ型）的抑制作用。结果GC3—1—4半数中毒浓度（TQ50）为144．17μg／ml；抑制副流感病毒（Ⅲ型）的半数有效浓度（EC50）为12．82μg／ml，治疗指数（TI）为11．25；GC3—1—4对副流感病毒（Ⅲ型）的抑制作用存在量效反应关系；在感染后6h以内给药，GC3—1—4对副流感病毒（Ⅲ型）均有抑制作用。结论：GC3—1—4在Hela细胞中对副流感病毒（Ⅲ型）有抑制作用。     

桂枝挥发油抗甲型流感病毒作用

目的:观察桂枝挥发油及其主要成分桂皮醛体外对甲型流感病毒A/PR/8/34(H1N1)增殖的影响及对该流感病毒株感染小鼠的治疗作用。方法:MTT法检测受试药物对狗肾传代细胞(MDCK)的半数中毒浓度(TC50)及最大无毒浓度(TC0);血凝法测定甲型流感病毒(H1N1)对MDCK细胞的感染性;MTT法和细胞病变法(CPE)测定两种药物体外抑制流感病毒增殖的有效浓度(IC50)和治疗指数(TI)。计算肺指数及HE染色观察药物对流感病毒(H1N1)感染小鼠的治疗作用。结果:桂枝挥发油和桂皮醛对MDCK细胞的TC50分别为5.38×10-3mg/ml和5.49×10-3mg/ml,TC0均为2.5×10-3mg/ml。桂枝挥发油和桂皮醛能抑制流感病毒在MDCK细胞内的增殖,其IC50分别为5.80×10-5mg/ml与5.31×10-5mg/ml,TI分别为92.82和103.35。桂枝挥发油0.174mg/kg和桂皮醛0.132mg/kg连续灌胃5d,明显降低病毒感染小鼠的肺指数,抑制率分别为26.7%和27.4%,并对病理组织形态有改善作用。结论:桂枝挥发油与桂皮醛体外明显抑制甲型流感病毒(H1N1)在MDCK细胞中的增殖,并对流感病毒株感染小鼠有较好的治疗作用。表明桂枝挥发油及桂皮醛具有抗甲型流感病毒作用,桂皮醛是桂枝挥发油抗病毒效应的主要有效成分之一。     

Spasmolytic activity of Toddalia asiatica Var. floribunda

The spasmolytic activity of the aerial parts of Toddalia asiatica var. floribunda (family Rutaceae) was evaluated. The ethanol extract exhibited significant spasmolytic activity and was then partitioned into five fractions. The activity was found to be concentrated only in the hexane and chloroform fractions. This activity was shown not to be due to the coumarins, toddalolactone and toddanone, as was previously thought.     

Antimicrobial activity of Ulopterol isolated from Toddalia asiatica (L.) Lam.: a traditional medicinal plant

Ethnopharmacological relevance

The leaves of Toddalia asiatica (L.) Lam. (Rutaceae) are widely used in folk medicine in India to treat various ailments like cough, malaria, indigestion, influenza lung diseases and rheumatism, fever, stomach ailments, cholera and diarrhea. In our earlier communication we have reported the antimicrobial study on the various extracts of the leaves and the isolation and identification of Flindersine, a quinolone alkaloid as the major active principle. In the present study, we report the antibacterial and antifungal activities of Ulopterol, a coumarin isolated as another major active antimicrobial principle.

Materials and methods

The leaves were successively extracted with hexane, chloroform, ethyl acetate, methanol and water. The extracts were studied for their antimicrobial activity against selected bacteria and fungi by using disc-diffusion method. The ethyl acetate extract which was found to possess highest antimicrobial activity was subjected to activity guided fractionation by column chromatography over silica gel. This resulted in the isolation of the coumarin, Ulopetrol, an active principle besides Flindersine which was reported by us earlier. The structure of the compound was elucidated using physical and spectroscopic data. Flindersine and Ulopterol were quantified by HPLC.

Results

Ulopterol showed activity against the bacteria viz. Staphylococcus epidermidis, Enterobacter aerogenes, Shigella flexneri, Klebsiella pneumoniae (ESBL-3967), Escherichia coli (ESBL-3984) and fungi viz. Aspergillus flavus, Candida krusei and Botrytis cinerea. Quantification by HPLC showed the content of Flindersine and Ulopterol to be 0.361% and 0.266% respectively on dry weight basis of the leaves.

Conclusions

Ethyl acetate extract (successive extraction) contained Ulopterol, a coumarin, besides Flindersine, a quinolone alkaloid, as a major active principle in the antimicrobial studies. This is the first report of the antimicrobial activity of Ulopterol and also its first report from the plant.     

In vitro antiviral activity of Phyllanthus orbicularis extracts against herpes simplex virus type 1

The antiviral activity of butanol- and acetic acid-soluble fractions, prepared from the leaves and stems of Phyllanthus orbicularis H.B.K., has been investigated against acyclovir-sensitive or -resistant herpes simplex virus type 1 (HSV-1) strains, using human foreskin fibroblast (HFF) and green ape kidney (Vero) cell lines. Both fractions showed antiviral selectivity indexes (SI) from 10.3 to 22.8, while their extracellular virucidal activities reached SI values ranging from 371 to 1,040. Time-addition experiments suggested that the active compounds present in the studied fractions acted on early steps of the virus replication cycle.     

Antiviral activity of puerarin as potent inhibitor of influenza virus neuraminidase

Puerarin is a major isofiavone compound isolated from the root of Pueraria lobata. It was reported that puerarin had antioxidant, antiinflammatory, antitumor, cholesterol lowering, liver protective, and neuroprotective properties. However, few studies have explored the antiviral effect of puerarin and its target mechanism related to influenza virus. Here, the antiinfluenza activity of puerarin in vitro and in vivo and its mode of action on the potential inhibition of neuraminidase (NA) were investigated. Puerarin displayed an inhibitory effect on A/FM/1/1947(H1N1) (EC₅₀ = 52.06 μM). An indirect immunofluorescence assay indicated that puerarin blocked the nuclear export of viral NP. The inhibition of NA activity confirmed that puerarin can block the release of newly formed virus particles from infected cells. Puerarin (100 and 200 mg/kg/d) exhibited effective antiviral activity in mice, conferring 50% and 70% protection from death against H1N1, reducing virus titers, and effectively alleviating inflammation in the lungs. The molecular docking results showed that puerarin had a strong binding affinity with NA from H1N1. The results of the molecular dynamics simulation revealed that puerarin had higher stable binding at the 150‐loop region of the NA protein. These results demonstrated that puerarin acts as a NA blocker to inhibit influenza A virus both in cellular and animal models. Thus, puerarin has potential utility for the treatment of the influenza virus infection.     

热毒宁注射液体外抑制甲型H1N1流感病毒的研究

目的：研究热毒宁注射液体外抑制甲型H1N1流感病毒的作用。方法以奥司他韦为阳性对照,采用CPE和MTT法观察热毒宁注射液对甲型H1N1流感病毒的抑制作用。结果 CPE法结果表明热毒宁注射液最大无毒浓度（TC0）为16.2 mg/mL,半数中毒浓度为（TC50）为（24.5±8.1）mg/mL;MTT法测定结果表明热毒宁注射液TC0为16.2 mg/mL,TC50为（21.7±9.4）mg/mL。热毒宁注射液体外抑制甲型H1N1流感病毒结果显示,CPE法和MTT法热毒宁注射液作用感染细胞1次组半数有效浓度（IC50）为（900.0±173.2）、（933.3±57.7）μg/mL,治疗指数（TI）为27.2、23.2;热毒宁注射液作用感染细胞3次组IC50为（666.7±115.5）、（866.7±208.1）μg/mL,TI为36.7、25.0。结论热毒宁注射液具有明显体外抗甲型H1N1流感病毒的作用。     

Evaluation of antiviral activity of South American plant extracts against herpes simplex virus type 1 and rabies virus

This paper describes the screening of different South American plant extracts and fractions. Aqueous and organic extracts were prepared and tested for antiherpetic (HSV-1, KOS and 29R strains) and antirabies (PV strain) activities. The evaluation of the potential antiviral activity of these extracts was performed by using an MTT assay for HSV-1, and by a viral cytopathic effect (CPE) inhibitory method for rabies virus (RV). The results were expressed as 50% cytotoxicity (CC(50)) for MTT assay and 50% effective (EC(50)) concentrations for CPE, and with them it was possible to calculate the selectivity indices (SI = CC(50)/EC(50)) of each tested material. From the 18 extracts/fractions tested, six extracts and four fractions showed antiviral action. Ilex paraguariensis, Lafoensia pacari, Passiflora edulis, Rubus imperialis and Slonea guianensis showed values of SI > 7 against HSV-1 KOS and 29-R strains and Alamanda schottii showed a SI of 5.6 against RV, PV strain.