姜黄素对5种非白念珠菌菌丝及生物膜形成的抑制作用

目的观察姜黄素对5种非白念珠菌菌丝及生物膜形成的抑制作用。方法测定姜黄素对热带念珠菌Candidatropicalis、光滑念珠菌C.glabrata、近平滑念珠菌C.parapsilokis、克柔念珠菌C.Krusei、季也蒙念珠菌C.guilliermondii的最小抑菌浓度(MIC)以及抑制50%生物膜浓度(SMIC50),利用光学显微镜观察姜黄素对非白念珠菌菌落形态的影响,利用倒置显微镜与扫描电镜分别观察姜黄素对非白念珠菌菌丝形成以及生物膜形成的影响。结果姜黄素对热带念珠菌、光滑念珠菌、近平滑念珠菌、克柔念珠菌、季也蒙念珠菌的MIC分别为64、128、256、256、128μg/m L;SMIC50分别为512、512、512、512、512μg/m L;倒置显微镜与扫描电镜观察发现姜黄素对5种非白念珠菌菌丝以及生物膜的形成均有抑制作用。光学显微镜观察发现姜黄素能抑制这5种非白念株菌在固体培养基上菌落周边菌丝的形成。结论姜黄素对5种非白念珠菌菌丝及生物膜的形成具有抑制作用。     

黄芩苷体外抗白念珠菌生物膜作用的研究

目的研究黄芩苷对体外白念珠菌生物膜的影响。方挂采用XTT减低法评价黄芩苷对白念珠菌的生物膜及黏附性的影响；镜下观察该药对白念珠菌生物膜的形态学影响；细胞毒试验检测该药的毒副作用。结果 各黄芩苷对白念珠菌生物膜的SMIC50、SMIC80分别是1000μg／ml、2000μg／ml;2000μg／ml及200μg／ml的黄芩苷对白念珠菌的早期黏附及菌丝生长有抑制作用；黄芩苷对人细胞毒性较弱。结论 黄芩苷对体外白念珠菌生物膜有较强的抑制作用。     

黄苓苷体外抗白念球菌生物膜作用的研究

目的 研究黄芩苷对体外白念珠菌生物膜的影响.方法 采用XTT减低法评价黄芩苷对白念珠菌的生物膜及黏附性的影响;镜下观察该药对白念珠菌生物膜的形态学影响:细胞毒试验检测该药的毒副作用.结果 各黄芩苷对白念珠菌生物膜的SMIC50、SMIC80分别是1 000μg/ml、2 000 μg/ml;2 000μg/ml及200μg/ml的黄芩苷对白念珠菌的早期黏附及菌丝生长有抑制作用:黄芩苷对人细胞毒性较弱.结论 黄芩苷对体外白念珠菌生物膜有较强的抑制作用.     

黄芩苷、黄芩素对变形链球菌生物膜抑制作用的研究

目的探讨黄芩苷、黄芩素对变形链球菌生物膜形成的影响,为中药生物膜抑制剂的研究开发提供一定的基础。方法以变形链球菌10556为研究对象,以氯已定为阳性对照药,测试黄芩苷、黄芩素对变形链球菌的最低抑制浓度(MIC);采用倍比稀释法对生物膜内活菌菌落计数;以原子力显微镜和扫描电镜观察其对生物膜形态结构的影响;结晶紫染色法测定对生物膜的抑制率。结果形态学观察可见生物膜被破坏,基质样物变稀疏,细菌群聚大为降低;同时,黄芩苷、黄芩素对变形链球菌生物膜形成具有明显的抑制作用。结论黄芩苷、黄芩素具有明显的体外抑菌作用,该结果可为中药黄芩抗菌作用的物质基础研究提供依据。     

黄芩苷体外对白念珠菌凋亡的影响

目的探讨中药有效成分黄芩苷对白念珠菌凋亡的影响。方法采用Hoechst33258染色荧光显微镜检测白念珠菌细胞凋亡的形态;Rh123染色流式细胞仪检测细胞线粒体膜电位(mitochondrial membrane potential,MMP)变化;DHR染色流式细胞仪检测细胞内活性氧(reactive oxygen species,ROS)水平。结果 1 000,100μmol/L的黄芩苷能诱导白念珠菌细胞核固缩与碎裂,并降低白念珠菌MMP;1 000,100,10μmol/L的黄芩苷均能提高细胞内ROS水平。结论黄芩苷可通过线粒体途径诱导白念珠菌凋亡。     

没食子酸抑制白念珠菌生物膜作用的研究

目的:研究没食子酸对体外白念珠菌生物膜的影响.方法:采用XTT减低法评价没食子酸对白念珠菌的生物膜及黏附性的影响;镜下观察没食子酸对白念珠菌生物膜的形态学影响;细胞毒试验检测该药的毒副作用.结果:没食子酸抑制白念珠菌生物膜最低药物浓度SMIC_(50),SMIC_(80)分别是500,1 000 mg·L~(-1);100,1 000 mg·L~(-1) 的没食子酸对白念珠菌的早期黏附及菌丝生长有抑制作用;没食子酸对人细胞毒性较弱.结论:没食子酸对体外白念珠菌生物膜有较强的抑制作用.     

黄芩苷、黄芩素对变形链球菌生物膜抑制作用的研究北大核心CSCD

目的探讨黄芩苷、黄芩素对变形链球菌生物膜形成的影响,为中药生物膜抑制剂的研究开发提供一定的基础。方法以变形链球菌10556为研究对象,以氯已定为阳性对照药,测试黄芩苷、黄芩素对变形链球菌的最低抑制浓度(MIC);采用倍比稀释法对生物膜内活菌菌落计数;以原子力显微镜和扫描电镜观察其对生物膜形态结构的影响;结晶紫染色法测定对生物膜的抑制率。结果形态学观察可见生物膜被破坏,基质样物变稀疏,细菌群聚大为降低;同时,黄芩苷、黄芩素对变形链球菌生物膜形成具有明显的抑制作用。结论黄芩苷、黄芩素具有明显的体外抑菌作用,该结果可为中药黄芩抗菌作用的物质基础研究提供依据。     

黄芩苷对白色念珠菌酶活性的影响

目的运用分光光度法检测黄芩苷作用白色念珠菌后,其琥珀酸脱氢酶(SDH)、细胞色素C氧化酶(CCO)、Ca2 -Mg2 -ATPase活性的改变,以探讨黄芩苷抗白色念珠菌作用靶位,为探索中药活性成分抗真菌作用机理提供依据。方法收集经黄芩苷作用了相应时间的白色念珠菌,制备菌悬液,反复冻融研磨,差速离心法提取白色念珠菌线粒体;Lowry法检测线粒体蛋白质含量;分光光度法检测SDH酶、CCO酶、Ca2 -Mg2 -ATPase活力。结果①0.25,0.5,1mg/ml黄芩苷浓度组白色念珠菌SDH活力明显低于0mg/ml黄芩苷浓度组(P<0.05),且出现黄芩苷浓度越高SDH活力越低的趋势。不同作用时间组间白色念珠菌SDH活力差异无统计学意义(P>0.05)。②白色念珠菌经不同浓度黄芩苷作用不同时间后,其CCO酶活力均无明显差异(P>0.05)。③0.25,0.5,1mg/ml黄芩苷浓度组与0mg/ml黄芩苷浓度组比较,Ca2 -Mg2 -ATPase活力差异显著,黄芩苷浓度越高,Ca2 -Mg2 -ATPase活力越低,作用时间对Ca2 -Mg2 -AT-Pase活力没有明显影响(P>0.05)。结论①黄芩苷具有明显的抗白色念珠菌作用;②黄芩苷可降低白色念珠菌SDH酶活力;③黄芩苷对白色念珠菌CCO酶活力未见明显影响;④黄芩苷可降低白色念珠菌Ca2 -Mg2 -ATPase活力。     

黄芩苷抗生物膜作用研究进展

随着抗生素耐药率的不断上升,加速了后抗生素时代的到来,生物膜的形成是细菌/真菌产生多药耐药的主要原因之一.黄芩苷是黄芩的主要活性成分,具有抗病毒、抗菌、抗过敏、抗肿瘤、保肝、利胆、免疫调节等多种药理作用,尤以抗真菌/细菌作用最为显著.大量研究表明黄芩苷可抑制生物膜的形成和发育,本文将近年来对黄芩苷抗生物膜作用及其作用机...     

黄芩苷对白念珠菌芽管形成及黏附性的影响

目的:研究黄芩苷对白念珠菌芽管形成、对白念珠菌黏附人口腔上皮细胞及阴道上皮细胞的影响.方法:将100,50,10 mg·L-1黄芩苷分别与白念珠菌悬液、白念珠菌与口腔上皮细胞混合液、白念珠菌与阴道上皮细胞混合液共同孵育,显微镜下计数芽管形成率、白念珠菌对口腔上皮细胞及阴道上皮细胞的黏附百分率.结果:100,50,10 mg·L-1黄芩苷均能抑制白念珠菌芽管形成及对口腔上皮细胞和阴道上皮细胞的黏附;白念珠菌标准株与临床株之间无显著性差异.结论:黄芩苷对体外白念珠菌的芽管形成及黏附性有较强抑制作用.     

In vitro activity of Tridax procumbens against promastigotes of Leishmania mexicana

Ethnopharmacological relevance

Tridax procumbens is an active herb against leishmaniasis.

Aim of the study

Leishmaniasis is a group of diseases caused by Leishmania protozoa. We investigated the antileishmanial activity of Tridax procumbens extracts and a pure compound against promastigotes of Leishmania mexicana, the causative agent of cutaneous leishmaniasis in the New World.

Materials and methods

Extracts and (3S)-16,17-didehydrofalcarinol (1) were obtained by chromatographic methods from Tridax procumbens, and the latter identified by spectroscopic analysis. The effect of these extracts and 1 on the growth inhibition of promastigotes of Leishmania mexicana was evaluated. In order to test the safety of extracts and 1, mammalian cells were treated with them, and cell viability was assessed using trypan blue and MTT.

Results

We demonstrated that extracts of Tridax procumbens and 1 showed a pronounced activity against Leishmania mexicana. The methanol extract inhibited promastigotes growth of Leishmania mexicana with a 50% inhibitory concentration (IC₅₀) of 3 μg/ml, while oxylipin 1 exhibited the highest inhibition at IC₅₀ = 0.478 μg/ml.

Conclusions

In this study we report the biological activity of extracts and (3S)-16,17-didehydrofalcarinol (1), obtained from Tridax procumbens, on the promastigote form of Leishmania mexicana, with no effect upon mammalian cells.     

In vitro antigonococcal activity of Coscinium fenestratum stem extract

Ethnopharmacological relevance

Gonorrhea is a sexually transmitted disease (STD), which originates from bacteria, Neisseria gonorrhoeae. It is still one of the major health problems worldwide in both men and women. Many medicinal plants have been recorded in Thai folk medicine for relieving STD but there is no scientific report of these plants for antigonococcal activity.

Aim of the study

This study was conducted to evaluate antigonococcal activity of 22 Thai medicinal plants selected from the plants popularly used in Thai folk medicine for treatment of gonorrhea.

Materials and methods

This study was performed by disc diffusion and agar dilution methods against Neisseria gonorrhoeae. Active compound was investigated by bioautographic assay.

Results

Among the selected plants, Coscinium fenestratum (Gaertn.) Colebr. extract showed the most effective activity against Neisseria gonorrhoeae ATCC 49226 with MIC value of 47.39 μg/ml. Bioautographic assay revealed that berberine was the active compound of Coscinium fenestratum against Neisseria gonorrhoeae. The average MIC values of purified berberine against Neisseria gonorrhoeae ATCC 49226 and 11 clinical isolates were 13.51 and 17.66 μg/ml, respectively while average MIC value of the crude extract of Coscinium fenestratum against all clinical isolates was about 56.39 μg/ml. There was no acute toxicity detected at the dose of 5 g of Coscinium fenestratum crude extract per kilogram.

Conclusions

These results provide theoretical support for ethnopharmacological relevance of antigonococcal activity of Coscinium fenestratum and its active compound.     

In vitro cytotoxic activity of nine plants used in Mayan traditional medicine

Ethnopharmacological relevance

Plants have been used in folk medicine by Mayan ancient people from the Yucatan Peninsula, Mexico, to treat some diseases considered as cancer diseases such as chronic wounds or tumors.

Aim of the study

We collected a selection of nine plants in order to investigate their cytotoxic activity against cancer cell lines.

Materials and methods

Methanolic extracts were tested for their cytotoxicity using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay on four cancer cell lines; nasopharynx carcinoma (KB), laryngeal carcinoma (Hep-2), cervix adenocarcinoma (HeLa), and cervix squamous carcinoma cells (SiHa) and one normal cell line; canine kidney (MDCK).

Results

All species exhibited some degree of cytotoxic activity. The root bark extract of Hamelia patens exhibited the highest cytotoxic activity on HeLa cells with a CC₅₀ of 13 μg/mL and selectivity index of 13.3, higher than docetaxel. Gossypium schottii and Dioon spinulosum showed similar good cytotoxic activity and selectivity index on HeLa and Hep-2 cells, respectively.

Conclusions

Hamelia patens, Dioon spinulosum and Gossypium schottii demonstrated promising cytotoxic activity and have been selected for future bio-guided fractionation and isolation of active cytotoxic compounds.     

In vitro and in vivo antimalarial and cytotoxic activity of five plants used in congolese traditional medicine

Aim of the study

The in vitro antiplasmodial activity and cytotoxicity of methanolic and dichloromethane extracts from five Congolese plants were evaluated. The plants were selected following an ethnobotanical survey conducted in D.R. Congo and focusing on plants used traditionally to treat malaria. The in vivo antimalarial activity of aqueous and methanolic extracts active in vitro was also determined in mice infected by Plasmodium berghei berghei.

Materials and methods

The growth inhibition of Plasmodium falciparum strains was evaluated using the measurement of lactate dehydrogenase activity. The extracts (aqueous, CH₃OH, EtOH and CH₂Cl₂) were prepared by maceration and tested in vitro against the 3D7 (chloroquine sensitive) and W2 (chloroquine resistant) strains of Plasmodium falciparum and against the human normal fetal lung fibroblasts WI-38 to determine the selectivity index. Some extracts were also used at the dose of 300 mg/kg to evaluate their activity in mice infected since 4 days by Plasmodium berghei.

Results

Two plants presented a very high activity (IC₅₀ < 3 μg/ml). These plants were Strychnos icaja roots bark (MeOH and CH₂Cl₂) and Physalis angulata leaves (MeOH and CH₂Cl₂). One plant (Anisopappus chinensis whole plant, MeOH and CH₂Cl₂) presented a high activity (IC50 < 15 μg/ml). The extracts of Anisopappus chinensis and Physalis angulata showed also a good inhibition of parasitemia in vivo. Flavonoids, phenolic acids and terpenes were identified in these plants by a general phytochemical screening method.

Conclusion

Three plants showed a very interesting antiplasmodial activity (Anisopappus chinensis, Physalis angulata and Strychnos icaja) and one of them showed a good selectivity index (>10, Anisopappus chinensis). Anisopappus chinensis and Physalis angulata were also active in vivo.     

In vitro antimicrobial activity of extracts and compounds of some selected medicinal plants from Cameroon

Aim of the study

Seven extracts and eight compounds from four selected Cameroonian medicinal plants, Solanecio mannii Hook f. (Asteraceae), Monodora myristica Dunal (Annonaceae), Albizia gummifera (J.F. Gmel) C.A. Smith (Fabaceae/Mimosoideae) and Glyphaea brevis (Spreng) Monachino (Tiliaceae), traditionally used for the treatment of hepatitis, parasites and other infectious diseases, were tested in vitro for their antimicrobial activity against Gram-positive (5 species) and Gram-negative (4 species) bacteria species and pathogenic yeasts (2 Candida species), to establish whether or not they have antimicrobial activity and to validate scientifically their use in traditional medicine.

Materials and methods

The agar disc diffusion and the microbroth dilution methods were used to determine the zone of inhibition between the edge of the filter paper and the edge of the inhibition area (IZ) and the minimal inhibitory concentration (MIC) respectively.

Results

The most active extracts against Candida albicans and Candida krusei were respectively the cyclohexane extract from the fruits of Monodora myristica and the ethyl acetate extract from the stem bark of Albizia gummifera (MIC = 6.3 μg/ml for both extracts). The lowest MIC value (1.6 μg/ml) for purified compounds was obtained on Candida albicans with a mixture of linear aliphatic primary alcohols (n-C24H50O to n-C30H62O), with n-hexacosanol (1b) as major compound and mixture of fatty acid esters of diunsaturated linear 1,2-diols (6).

Conclusion

These results afford ground informations for the potential use of the crude extracts of these species as well as of some of the isolated compounds in bacterial and fungal infections.     

In vitro antimicrobial activity of Caesalpinia ferrea Martius fruits against oral pathogens

Aim

In the Amazon region of Brazil, the fruits of Caesalpinia ferrea Martius (Brazilian ironwood) are widely used as an antimicrobial and healing medicine in many situations including oral infections. This study aimed to evaluate the antimicrobial activity of Caesalpinia ferrea Martius fruit extract against oral pathogens.

Materials and methods

Polyphenols estimation and spectral analysis (¹H NMR) of the methanol extract were carried out. The microorganisms Candida albicans, Streptococcus mutans, Streptococcus salivarius, Streptococcus oralis and Lactobacillus casei were tested using the microdilution method for planktonic cells (MIC) and a multispecies biofilm model. Chlorhexidine was used as positive control.

Results

Polyphenols in the extract were estimated at 7.3% and ¹H NMR analysis revealed hydroxy phenols and methoxilated compounds. MIC values for Candida albicans, Streptococcus mutans, Streptococcus salivarius, Streptococcus oralis and Lactobacillus casei were 25.0, 40.0, 66.0, 100.0, 66.0 μg/mL, respectively. For the biofilm assay, chlorhexidine and plant extract showed no growth at 10⁻⁴ and 10⁻⁵ microbial dilution, respectively. At 10⁻⁴ and 10⁻⁵ the growth values (mean ± SD) of the negative controls (DMSO and saline solution) for Streptococcus mutans, Streptococcus sp. and Candida albicans were 8.1 ± 0.7, 7.0 ± 0.6 and 5.9 ± 0.9 × 10⁶ CFU, respectively.

Conclusion

Caesalpinia ferrea fruit extract can inhibit in vitro growth of oral pathogens in planktonic and biofilm models supporting its use for oral infections.     

巫山淫羊藿离体胚培养的研究

目的:研究药用植物巫山淫羊藿组织培养技术,为工厂化育苗提供科学根据。方法:以巫山淫羊藿的种胚为外植体,采用MS培养基,附加不同浓度2,4-D,6-BA,IBA,NAA进行正交试验。结果:诱导愈伤组织的最优培养基为:MS+2,4-D2 mg.L-1+IBA 2 mg.L-1+NAA 0.5 mg.L-1;愈伤组织分化的最优培养基为:MS+6-BA 1 mg.L-1+NAA 0.5 mg.L-1+IBA 1 mg.L-1;诱导芽的最优培养基为:MS+IBA 2 mg.L-1+6-BA 0.5 mg.L-1;芽增殖的最佳培养基为:MS+6-BA 1.0 mg.L-1+NAA 0.5 mg.L-1。结论:通过诱导愈伤组织途径和丛生芽途径,建立了巫山淫羊藿种胚外植体诱导和培养方法,达到快速繁殖的目的。     

In vivo antimalarial activity of Keetia leucantha twigs extracts and in vitro antiplasmodial effect of their constituents

Ethnopharmacological relevance

The West African tree Keetia leucantha (Rubiaceae) is used in traditional medicine in Benin to treat malaria. The twigs dichloromethane extract was previously shown to inhibit in vitro Plasmodium falciparum growth with no cytotoxicity (>100 µg/ml on human normal fibroblasts).

Materials and methods

The dichloromethane and aqueous extracts of twigs of K. leucantha were evaluated in vivo against Plasmodium berghei NK 173 by the 4-day suppressive test and in vitro against a chloroquine-sensitive strain of Plasmodium falciparum (3D7) using the measurement of the plasmodial lactate dehydrogenase activity. Bioguided fractionations were realized and compounds were structurally elucidated using extensive spectroscopic analysis.

Results

The in vivo antimalarial activity of K. leucantha dichloromethane and aqueous twigs extracts were assessed in mice at the dose of 200 mg/kg/day. Both extracts exhibited significant effect in inhibiting parasite growth by 56.8% and 53.0% (p<0.0001) on day 7-postinfection. An LC–MS analysis and bioguided fractionations on the twigs dichloromethane extract led to the isolation and structural determination of scopoletin (1), stigmasterol (2), three phenolic compounds: vanillin (3), hydroxybenzaldehyde (4) and ferulaldehyde (5), eight triterpenic esters (6–13), oleanolic acid and ursolic acid. The antiplasmodial activity of the mixture of the eight triterpenic esters showed an antiplasmodial activity of 1.66±0.54 µg/ml on the 3D7 strain, and the same range of activity was observed for isolated isomers mixtures.

Conclusions

This is the first report on the in vivo activity of K. leucantha extracts, the isolation of thirteen compounds and analysis of their antiplasmodial activity. The results obtained may partially justify the traditional use of K. leucantha to treat malaria in Benin.     

In vitro estrogenic activity of Asplenium trichomanes L. extracts and isolated compounds

Ethnopharmacological relevance

Asplenium trichomanes was used as an expectorant, anti-cough remedy, laxative, emmenagogue, abortifacient and for irregular menses.

Aim of the study

To investigate the in vitro estrogenic activity of Asplenium trichomanes extracts and isolated compounds and their ability to activate ERα and ERβ.

Materials and methods

Leaves infusion (IF), decoction (DC) and methanol extract (ME) were prepared. MCF7/EREluc cell line which expresses endogenous ERα, and SK-NBE cells transiently transfected with the estrogen receptors (ERα and ERβ) were used for the estrogenic activity assays. Phytochemical investigations were performed (CC, HPLC, etc.) and structure of isolated compounds were achieved on the basis of 1D and 2D NMR techniques and HR-MS spectrometry.

Results

IF and ME were active in our MCF7 model; selectivity for the ERβ receptor was observed in the SK-NBE test. Two new phenol derivatives, 4-vinyl-phenol-1-O-[α-l-rhamno(1 → 6)-β-d-glucopyranosyde] (1) and kaempferol-3-O-α-[2′acetyl]-arabinofuranosyl-7-O-α-l-rhamnopyranoside (2) were isolated with six known compounds (3–8). Compounds 2–4, 7 and 8 showed selectivity for the activation of the ERβ receptor although with a moderate activity compared with 17-β-estradiol.

Conclusion

Further investigations about the estrogenic effects of this plant are needed but our data can, at least in part, explain some of its traditional use as emmeagogue.     

In vitro antiplasmodial activity of plants used in Benin in traditional medicine to treat malaria

Aim of the study

The aim of the study was to evaluate the in vitro antiplasmodial activity of crude extracts of 12 plant species traditionally used in Benin for the treatment of malaria in order to validate their use.

Materials and methods

For each species, dichloromethane, methanol and total aqueous extracts were tested. The antiplasmodial activity of extracts was evaluated using the measurement of the plasmodial lactate dehydrogenase activity on chloroquine-sensitive (3D7) and resistant (W2) strains of Plasmodium falciparum. The selectivity of the different extracts was evaluated using the MTT test on J774 macrophage-like murine cells and WI38 human normal fibroblasts.

Results

The best growth inhibition of both strains of Plasmodium falciparum was observed with the dichloromethane extracts of Acanthospermum hispidum DC. (Asteraceae) (IC₅₀ = 7.5 μg/ml on 3D7 and 4.8 μg/ml on W2), Keetia leucantha (K. Krause) Bridson (syn. Plectronia leucantha Krause) (Rubiaceae) leaves and twigs (IC₅₀ = 13.8 and 11.3 μg/ml on 3D7 and IC₅₀ = 26.5 and 15.8 μg/ml on W2, respectively), Carpolobia lutea G.Don. (Polygalaceae) (IC₅₀ = 19.4 μg/ml on 3D7 and 8.1 μg/ml on W2) and Strychnos spinosa Lam. (Loganiaceae) leaves (IC₅₀ = 15.6 μg/ml on 3D7 and 8.9 μg/ml on W2). All these extracts had a low cytotoxicity.

Conclusion

Our study gives some justifications for the traditional uses of some investigated plants.