贵州老鹰茶提取物体外抗单纯疱疹病毒1型活性的实验研究

目的研究贵州老鹰茶甲醇提取物体外抗单纯疱疹病毒1型(HSV-1)活性。方法采用细胞病变效应(Cytopatho-genic effect,CPE)法与空斑减数实验(Plaque reduction assay,PRA)测定甲醇提取物抗HSV-1活性,计算其空斑抑制率和半数抑制浓度(50%inhibiting concentration,IC50),并从药物对病毒的直接灭活作用、对病毒吸附的影响及对病毒穿膜的影响3个方面初步探讨其抗HSV-1的活性机理。结果贵州老鹰茶甲醇提取物能明显抑制HSV-1的致病变作用,其IC50为12.02μg/ml。药物主要是影响病毒对细胞的吸附,对病毒也有一定的直接灭活作用。结论贵州老鹰茶甲醇提取物有显著的抗HSV-1活性,初步推测其抗HSV-1活性的机理是作用在HSV-1和受体结合,侵入Vero细胞阶段。     

桉叶水提物体外抗单纯疱疹病毒Ⅰ型活性的实验研究

目的：研究直干桉叶水提物体外抗单纯疱疹病毒1型（HSV-1）活性。方法：采用CPE观察实验与空斑减数实验测定直干桉叶水提物抗HSV-1活性，计算其空斑抑制率和IC50，并从药物对病毒的直接灭活作用、对病毒吸附的影响及对病毒穿膜的影响3个方面初探直干桉叶水提物抗HSV-1活性机理。结果：直干桉叶水提物能明显抑制HSV-1的致病变作用，其IC50为84．28μg／ml。药物主要是对病毒有直接灭活作用，也能影响对病毒对细胞的吸附。结论：直干桉叶水提物有显著的抗HSV-1活性，初步推测直干桉叶水提物抗HSV-1活性的机理是影响病毒的囊膜结构从而使之失活。     

裙带菜茎硫酸多糖体外抗单纯疱病毒Ⅰ型活性的实验研究

目的 研究裙带菜茎多糖体外抗单纯疱疹病毒1型(HSV-1)活性.方法 采用空斑减数实验测定裙带菜茎粗多糖及纯化后多糖抗HSV-1活性,计算它们的空斑抑制率和IC50,并从药物对病毒的直接灭活作用、对病毒吸附的影响及对病毒释放的影响3个方面初探裙带菜茎多糖抗单纯疱疹病毒活性机理.结果 裙带菜茎多糖能明显抑制HSV-1的致病变作用,IC50依次为 8.04,13.54 μg/ml.药物对病毒没有直接灭活作用,对病毒的释放也没有影响,主要影响了病毒的吸附.结论 裙带菜茎多糖有显著的抗HSV-1活性,且粗多糖比纯化后多糖活性要好,并且可以初步推测裙带菜茎多糖抗HSV-1活性是作用在病毒和受体结合、侵入Vero细胞阶段.     

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。     

直杆桉果实提取物体外抗单纯疱疹病毒和乙型肝炎病毒的实验研究

目的:研究3种不同溶剂的直杆桉果实提取物体外抗单纯疱疹病毒Ⅰ型(HSV-1)和乙型肝炎病毒(HBV)的活性。方法:采用MTT法检测药物毒性,CPE法与空斑减数实验检测药物抗HSV-1活性;采用ELISA法检测药物对HepG2.2.15细胞分泌的乙型肝炎表面抗原(HBsAg)和乙型肝炎E抗原(HBeAg)的抑制作用。结果:直杆桉叶果实的醋酸乙酯提取物(P18-E1)和甲醇提取物(P18-E2)无明显抗HSV-1的活性,水提物(P18-E3)能明显抑制HSV-1的致病变作用,其TC50为69.20 mg/mLI,C50为126.77μg/mL,治疗指数为545.87;3种不同溶剂的提取物对HepG2.2.15细胞分泌的HBsAg和HBeAg均无明显抑制作用。结论:直杆桉果实的水提物(P18-E3)具有显著的抗HSV-1活性,主要是对病毒有直接灭活作用,也能影响病毒对细胞的吸附。初步推测其抗HSV-1活性的机理是影响病毒的囊膜结构从而使之失活。     

5种南药的19种提取物体外抗甲型流感病毒活性的实验研究

目的探讨南药提取物体外抗甲型流感病毒的活性。方法采用细胞病变效应法(CPE法)检测19种南药提取物在MDCK细胞中的抗甲型流感病毒的活性,并通过不同的加药方式初步探索具有活性的提取物抗甲型流感病毒活性的机理。结果 在19种提取物中,直杆桉果实水提物和川楝子醇提取物具有抗甲型流感病毒的活性,初步作用机制研究表明直杆桉果实水提物对甲型流感病毒具有直接灭活的作用,而川楝子提取物则具有明显的预防效果。总体上直杆桉果实提取物的抗甲型流感病毒的活性更显著。结论 直杆桉果实水提取物和川楝子醇提取物具有抗甲型流感病毒的活性,且直杆桉果实水提物抗H1N1的活性更加显著。     

大青叶有效单体抗呼吸道合胞病毒作用的实验研究

目的 探讨中药大青叶单体1#,2#,3#,4#体外抗呼吸道合胞病毒(RSV)的作用.方法 通过观察病毒引起的细胞病变效应(Cytopathogenic effect,CPE)和MTT[Methyl Thiazolyl Tetrazolium,3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐]法检测药物抗RSV活性, 计算药物对病变的抑制率和半数抑制浓度(50% inhibiting concentration ,IC50),并从药物抗病毒吸附,药物对病毒的直接杀灭作用及抑制病毒在细胞内的生物合成3个方面初步探索大青叶单体抗RSV活性的作用机制.结果 大青叶单体1#,2#,3#,4#对RSV无直接灭活作用,也不能阻止RSV的吸附,而能抑制RSV在Hep-2细胞内的的生物合成,其半数抑制浓度(IC50)分别为32.57,26.37,31.60,29.61 μg/ml,治疗指数(TI) 分别为6.15,10.50,9.21,13.22,其抗病毒效果优于病毒唑(IC50=103.20 μg/ml, TI=5.21)和抗病毒口服液(IC50=105.21 μg/ml, TI=5.03) ,P<0.01.结论 大青叶单体1#,2#,3#,4#能安全高效地抑制RSV在Hep-2细胞中的增殖,抑制作用发生在病毒入侵细胞后.     

藏药紫金标抗HSV-1的作用机理研究

目的 :探讨紫金标抗单纯疱疹Ⅰ型病毒 (HSV-1)的作用及机理 ,为进一步开发该药提供理论依据。方法 :采用不同剂量的紫金标作用于适量HSV-1感染的Vero细胞 ,以 50%组织细胞感染量 (TCID₅₀ ) ,细胞病变效应(CPE) ,MTT法和核酸分子杂交作为评价指标。结果 :MTT法测得紫金标的 50 %抑制浓度 (IC₅₀ )为 29.46mg·L^-1,50%中毒浓度 (TC₅₀ )为 1 077mg·L^-1,治疗指数 (TI)为 36.56 ,结果表明紫金标有明显抑制HSV-1的作用 ,其作用强度和有效时间与药物浓度成正比。紫金标无直接灭活HSV-1的作用 ,也不能影响病毒的释放 ;但可干扰HSV -1对宿主细胞的吸附。不同浓度的紫金标能明显抑制HSV-1gD基因复制和mRNA表达。结论 :紫金标具有显著的抗HSV-1的作用 ,能抑制HSV-1对宿主细胞的吸附以及抑制HSV-1gD基因复制与转录。     

金银花体外抗呼吸道合胞病毒作用研究

目的:研究金银花体外抑制呼吸道合胞病毒作用的效果和强度。方法:采用细胞病变抑制实验,噻唑蓝(MTT)比色法检测细胞活性,观察金银花在人宫颈癌传代细胞(Hela)中对人呼吸道合胞病毒3型的抑制作用,以治疗指数(TI)为评价指标。结果:金银花对Hela细胞半数中毒浓度(TC50)为5 mg/mL,最大无毒浓度(TC0)为3.6 mg/mL。对呼吸道合胞病毒有直接灭活作用,其半数抑制率(IC50)为0.16 mg/mL,TI为31.2;在吸附阶段也有作用,其IC50为0.48 mg/mL,TI为10.5;同时金银花有抑制呼吸道合胞病毒生物合成作用,其IC50为1.0 mg/mL,TI为5.0;金银花不能阻止呼吸道合胞病毒侵入细胞。结论:金银花在体外主要通过直接灭活、阻止病毒吸附和抑制生物合成三种方式发挥抗呼吸道合胞病毒作用。     

藏药甘青乌头抗单纯疱疹病毒Ⅱ型体内外作用研究

 目的探讨藏药甘青乌头抗单纯疱疹病毒Ⅱ型(HSV-2)的作用和机制。方法MTT法测定甘青乌头对Vero细胞的毒性,采用细胞病变法和蚀斑法测定甘青乌头体外抗病毒活性,以半数抑制浓度和治疗指数为评价指标;定量荧光PCR法和蚀斑法考察甘青乌头体外抗病毒作用机制。用HSV-2建立小鼠脑炎模型,对甘青乌头体内抗单纯疱疹病毒的抗病毒活性进行评价。结果甘青乌头对Vero细胞的半数中毒浓度(CC₅₀)为5.57 g·L^-1。细胞病变法和蚀斑法测得甘青乌头的半数有效浓度(EC₅₀)分别为2.25,1.68 g·L^-1,治疗指数TI分别为2.47,3.32。LD₅₀值为0.85 g·kg^-1。甘青乌头延长了小鼠的平均存活时间,对小鼠的死亡显示出10%的保护率。甘青乌头能直接灭活HSV-2,能够显著抑制HSV-2的感染性;对病毒吸附到细胞没有抑制作用,能抑制HSV-2大分子的增值。抑制HSV-2 DNA合成的抑制倍数达102。结论甘青乌头体外具有较强的抗HSV-2的活性,抗病毒作用是通过抑制病毒复制循环的各个环节起作用的。     

壁虎醇提物对人食管鳞癌细胞EC9706的作用和体内抗肿瘤活性

目的:观察壁虎醇提物体内外抗肿瘤作用。方法:体外实验采用四噻唑蓝法(MTT)检测壁虎醇提物对人食管鳞癌细胞EC9706的增殖抑制作用;采用Hoechst33258荧光染色观察细胞凋亡的形态改变;采用TUNEL法检测细胞凋亡率;采用SP免疫组化法检测细胞内凋亡蛋白Bax和Bc l-2的表达。体内实验以小鼠S180肉瘤为模型,观察壁虎醇提物对肿瘤生长的抑制作用。结果:6~8 g.L-1壁虎醇提物作用于细胞24,48,72 h后能够明显抑制EC9706食管鳞癌细胞株的生长(P(0.01),抑制作用呈剂量和时间依赖性;Hoechst33258荧光染色可见凋亡细胞;6,7 g.L-1壁虎醇提物处理组细胞凋亡率高于阴性对照组(P0.05,P0.01),凋亡率分别为(12.73±3.84)%,(19.80±2.32)%;免疫组化结果显示细胞内Bc l-2蛋白无明显变化,而Bax蛋白表达升高(P(0.01),Bax/Bc l-2升高。不同剂量的壁虎醇提物(0.6,1.2,2.4 g.kg-1)均可抑制S180肉瘤在昆明小鼠体内生长,抑瘤率分别为44.88%,63.94%,69.53%。结论:壁虎醇提物体内体外具有抗肿瘤作用,体外诱导EC9706细胞凋亡的作用机制可能与凋亡相关蛋白Bax/Bc l-2升高有关。     

白背叶提取物A的体外抗肿瘤活性研究

目的观察白背叶提取物A对肿瘤细胞增殖的抑制作用。方法采用四唑盐比色试验(MTT法),以半数抑制浓度(IC50)为评价抗肿瘤活性强度的指标,分别对HL60人白血病细胞、Hela人宫颈癌细胞、A375人黑色素瘤细胞、MCF7人乳腺癌细胞4种常见的人恶性肿瘤细胞进行抗肿瘤活性的筛选。结果白背叶提取物A在浓度为10μg/ml时对HL60(P<0.05)、Hela(P<0.05)、A375(P<0.05)、MCF7(P<0.01)即有明显抑制作用(P<0.05或P<0.01),其IC50分别为29.64,30.36,36.65,42.81μg/ml。结论白背叶提取物A对以上4种肿瘤细胞的增殖均有抑制作用。     

Genotoxicity testing of Cecropia obtusifolia extracts in two in vivo assays: the wing somatic mutation and recombination test of Drosophila and the human cytokinesis-block micronucleus test

Cecropia obtusifolia Bertol. (Cecropiaceae) is a tree that grows in secondary vegetation in the tropical rain forest along both coasts of Mexico. Its leaves are used in folk medicine for the treatment of diabetes mellitus type 2. The aim of the present studies was the evaluation of possible genotoxic effects of the aqueous extract from the leaves of Cecropia obtusifolia by means of two different experimental assay models: the wing somatic mutation and recombination test in flies and the micronucleus test from lymphocytes obtained from patients treated with the extract. No toxicity was found to be induced by the leaves of Cecropia obtusifolia. The Drosophila wing somatic mutation and recombination test (SMART) was applied in the standard version with basal biotransformation activity as well as in a variant version with increased cytochrome P450-dependent bioactivation capacity. The ranges of exposure concentrations for these genotoxicity experiments were between 0.82 and 13.32 mg/ml. The extract did not produce any genotoxic effect; however it showed a non significant antigenotoxic effect. The human micronucleus assay in vivo was performed with cultured lymphocytes obtained from six diagnosed type 2 diabetic patients treated daily with 13.5 g of the aqueous extract between 32 and 85 days. No statistically significant increases in cytotoxicity and/or genotoxicity between control and diabetic blood samples were observed.     

Effect of Bacopa monniera Linn. extract on murine immune response in vitro

The study was to investigate and compare the effects of the Bacopa monniera Linn. extract and bacoside A on the ICR mice immune system in vitro. Splenocyte proliferation without or with mitogen (lipopolysaccharide, pokeweed mitogen, phytohaemagglutinin and concanavalin A) and phagocytic activity were assayed. The results showed that B. monniera extract at 0.001-1 mg/mL slightly suppressed splenocyte proliferation (SI 0.7) and decreased T-lymphocyte proliferation (SI 0.4) at 0.001 and 0.1 mg/mL with concanavalin A. Bacoside A at 0.001 mg/mL gave the highest splenocyte proliferation (SI 1.5) and strongly increased T-lymphocyte proliferation (SI 2.0) at 0.1 mg/mL with concanavalin A. Thus, it is possible to attribute the effect of B. monniera extract on splenocyte proliferation to the presence of bacoside A with other combined components. However, only B. monniera extract at 10 mg/mL produced a slight increase in lysosomal enzyme activity (PI 1.2), indicating a weak effect on phagocytic activation. It might be concluded that B. monniera manifests various effects on the murine immune system depending on the immune cell types, in accordance with its folklore uses. New assays are being carried out to study its mechanisms and to further investigate its applications in the treatment of human immune mediated diseases.     

墨旱莲对香烟提取物杀伤NHBE细胞的保护作用及机制

目的:研究墨旱莲提取物对香烟烟雾提取物(cigarette smoke extract,CSE)作用的正常肺细胞NHBE细胞的保护作用及其机制。方法:MTT法检测墨旱莲提取物对CSE诱导的NHBE细胞增殖的影响,DTNB比色法测定GSH含量,CDNB比色法检测GST活力,NADPH与2,6-二氯靛酚钠(DCIP)测试NQO1活力,Western blot测定蛋白的表达。结果:墨旱莲提取物可以降低CSE对NHBE细胞增殖的抑制作用;恢复由CSE引起的细胞内GSH水平下降,降低诱导细胞中GST与NQO1活力及NQO1蛋白表达。结论:墨旱莲能够减少CSE对NHBE细胞的损伤,其机制可能与II相解毒酶相关。     

In vitro immunomodulatory effect of Pouteria cambodiana (Pierre ex Dubard) Baehni extract

The methanolic stem bark extract from Pouteria cambodiana (Pierre ex Dubard) Baehni was evaluated for immunomodulating activity on BALB/c mice. The antioxidant effect was also assessed. The extract presented a good dose-response effect in the peritoneal macrophage phagocytosis assay with higher activity at 1mg/ml and an EC50 of 0.02 mg/ml and also activated lysosomal enzyme activity with an EC50 of 0.16 mg/ml. In the splenocyte proliferation assay, the extract without mitogen was active (EC50, 0.01 mg/ml) while the EC50 of the extract with lipopolysaccharide (LPS) and pokeweed mitogen (PWM) were 0.02 and 0.41 mg/ml, respectively. The extract showed low free radical scavenging activity in the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical assay with an IC50 of 0.24 mg/ml, being less active than ascorbic acid, butylated hydroxytoluene (BHT) and alpha-tocopherol which showed IC50 of 0.08, 0.10 and 0.11 mg/ml, respectively. The extract at doses up to 0.073 mg/ml had no effect on lipid peroxidation. The potent immunological but no antioxidant activity of the extract presented in this study can explain, at least in part, the Thai folklore application of this plant in the treatment of fever and skin eruption.     

Genotoxic and antigenotoxic properties of Baccharis trimera in mice

Ethnopharmacological relevance

Baccharis trimera (Less.) (Asteraceae) is a native plant from Brazil. Also known as “carqueja”, it is popularly used to treat liver diseases, diabetes, as well as digestive disorders, mainly by women with lower socioeconomic status.

Aim of the study

The aim of the present study was to investigate the in vivo genotoxic/antigenotoxic and mutagenic potential of this plant, using the comet and the micronucleus assays.

Material and methods

Female adult mice were treated with 500, 1000 or 2000 mg/kg of B. trimera aqueous extract (Bt-AE) by gavage for three consecutive days.

Results

Independently of the dose, no genotoxic effect was observed in blood and liver samples analyzed by the comet assay. Conversely, B. trimera showed an antigenotoxic effect in blood from treated mice, thus protecting cells against oxidative DNA damage induced by the ex vivo treatment with hydrogen peroxide. In addition, Bt-AE showed in vitro antioxidant activity, assessed by DPPH and xanthine oxidase assays, suggesting that the observed antigenotoxic effects might be related to its antioxidant properties.

Conclusions

However, the extract increased the frequency of micronucleus in bone marrow of treated mice, indicating a chromosomal mutagenic activity. Thus, medicines prepared from this plant should be used with caution, although the results also suggest antigenotoxic effects for B. trimera aqueous extract.     

金钗石斛中联苄类和酚酸类成分的抗氧化活性研究

 目的研究金钗石斛体积分数60%乙醇提取物中联苄类和酚酸类成分的抗氧化活性,探讨其抗氧化活性与结构的关系。方法综合运用现代色谱学和光谱学方法对金钗石斛体积分数60%乙醇提取物中的化学成分进行分离鉴定;采用1,1-苯基苦基苯肼(DPPH)自由基清除法和氧自由基清除能力法两种体外活性测试方法评价分得的16个化合物的抗氧化活性。结果从金钗石斛体积分数60%乙醇提取物中分离得到3个新的联苄类化合物和13个酚酸类化合物,将它们的结构鉴定为nobilinA(1),nobilinB(2),nobilinC(3),丁香酸(syringicacid)(4),2-羟基苯丙醇(2-hydroxyphenylpropanol)(5),香草醛(vanil-lin)(6),罗布麻宁(apocynin)(7),松柏醛(coniferylaldehyde)(8),丁香醛(syringaldehyde)(9),丁香乙酮(syringylethanone)(10),对羟基苯甲醛(p-hydroxybenzaldehyde)(11),3-羟基-4-甲氧基苯乙醇(3-hydroxy-4-methoxyphenylethanol)(12),α-羟基丁香丙酮(α-hydroxysyringylethanone)(13),二氢松柏醇(dihydroxyconiferylalcohol)(14),对羟基苯甲酸(p-hydroxybenzoicacid)(15),对羟基苯丙酸(p-hydroxyphenylpropionicacid)(16)。1-二苯基-2-苄基苯肼(DPPH)法活性测试中,化合物4的活性明显强于维生素C(VitC),化合物1,2,12,14显示出弱于VitC但强于2,6-二羟丁基对甲酚(BHT)的活性,化合物3和8只显示很弱的活性;氧自由基清除能力法(ORAC)法测试结果表明,化合物2～4,7～10,15显示出强于VitC的活性,化合物12,14,16活性较弱。结论金钗石斛中的联苄类和酚酸类化合物大多具有不同程度的抗氧化活性,活性的强弱与分子结构之间存在一定的相关性。