Allelopathy of Cold Water Extracts from Origanum vulgare ssp. vulgare L.

Secondary metabolites in medicinal plants could lead to discovery of new classes of herbicides. Recently aromatic plants have gained interest as a source of allelopathic secondary metabolites. Origanum vulgare ssp. vulgare L. infusions in hot water are used in folk medicine and possess proved beneficial biological activity. Plant-to-plant variability of metabolites due to genetic heterogeneity is established in Lamiaceae family. From this point of view, studies on plants from different geographic regions might reveal important sources of variability. The objective of this study was to evaluate allelopathic activity of cold water extracts made from the aerial parts of O. vulgare ssp. vulgare growing wild in Northeast Bulgaria in laboratory conditions. The allelopathic effect was evaluated using root elongation test and Allium cepa-test. Oregano extracts (17.5 g/l, 52.5 g/l) significantly decreased root length of Triticum aestivum L. (P ≤ 0.001). The root growth reduction could serve as a sign for presence of water soluble allelopathic secondary metabolites in the plant tested. Oregano (3.5 g/l) inhibited cell division in Allium root meristematic cells. The decline of the mitotic index indicates the occurrence of a cytotoxic effect. Oregano induced abnormalities in mitotic and interphase cells, so can be also considered as genotoxic. The observed macroscopic and microscopic effects of tested extracts indicated presence of water soluble allelochemicals in O. vulgare ssp. vulgare. This characteristic could be further studied as a possibility to be used in weed management programs.

Origanum vulgare L.leaf extract alleviates finasteride-induced oxidative stress in mouse liver and kidney

Objective:To investigate the hepatorenoprotective effects of Origanum vulgare L.against finasteride-induced oxidative injury in the liver and kidney of mice.Methods:Liquid chromatography-electrospray ionization-tandem mass spectrometry(LC-ESI/MS)analysis was utilized to yield a fingerprint of Origanum vulgare polyphenolic constituents.Thirty BALB/c mice received 0.5 mL/day distilled water,finasteride(25 mg/kg/day for 10 d),and 100,200,or 400 mg/kg/day finasteride+Origanum vulgare extract with 6 mice per group for five weeks.On day 36,liver and kidney function as well as pro-and antiinflammatory(IFN-γ,IL-12,IL-6,TNF-α,IL-1β,and IL-10)cytokines were measured.The total antioxidant status,nitric oxide(NO),and malondialdehyde levels as well as the activities of NO synthase and catalase were also evaluated.Histopathological study was conducted to assess the effect of Origanum vulgare extract on finasteride-induced renal and hepatic toxicities.Results:Twenty-five major polyphenolic compounds were identified in the Origanum vulgare extract by LC-ESI/MS.Origanum vulgare extract,especially at 200 and 400 mg/kg/day doses,significantly improved liver and kidney biochemical indices,decreased inflammatory cytokines,increased total antioxidant status and NO synthase and catalase activities,as well as decreased plasma NO and malondialdehyde levels in a dose-dependent manner as compared to the finasteride group.Histopathological results further confirmed the protective effect of Origanum vulgare extract.Conclusions:Origanum vulgare extract ameliorates finasterideinduced hepatic and renal biochemical and histopathological alterations,and restores antioxidant/oxidant balance.

HPLC法测定牛至中迷迭香酸的含量

目的 建立牛至中迷迭香酸含量的HPLC测定方法 。方法 采用高效液相色谱法（HPLC）测定牛至中迷迭香酸的含量。色谱条件：Kromasil C18柱（4．6mm×150mm，5μm），流动相为甲醇-1．5％乙酸水（38：62），流速为1．0mL／min，检测波长为330nm，柱温25℃。结果 迷迭香酸峰面积与进样量在0．3321～1．6605μg范围内有良好的线性关系（r=0．9999），平均回收率为98．96％，RSD为1．09％（n=6）。10个产地的牛至中迷迭香酸含量为8．13-15．74mg／g。结论 HPLC法操作简单、准确可靠，可用于牛至中迷迭香酸含量的测定。

The Use of Restriction Fragment Length Polymorphism and Fluorescence in Situ Hybridization to Investigate Microbiota of Piglets after Feeding Oregano

A total of 80 piglets (7.9 ± 1.0 kg) were used in a feeding experiment with dried oregano. The diets differed in their oregano content: 0 g, 2 g, 4 g and 8 g oregano/kg feed, corresponding to 0, 23.5, 46.9 and 93.9 mg carvacrol/kg DM. After the experimental period of 5 weeks, 20 piglets of both extreme feeding groups were slaughtered: 10 animals of the control group and 10 animals of the group that received 8 g oregano/kg. Ingesta samples of jejunum, caecum and colon were collected and analyzed by FISH and PCR RFLP to compare the diversity of microbiota. The results showed no significant changes in microbiota in response to oregano. The patterns of the PCR-RFLP showed a similarity of 61.8% - 91.8% in both feeding groups. In conclusion, an effect of oregano on the in- testinal microbiota could not be shown under the methods used.

牛至中药组方汤剂治疗兔球虫病的疗效试验及效果评估

为探讨牛至中药组方对兔球虫病的治疗效果,该试验选用了25只体重相近的45日龄健康无球虫试验兔,并随机分成5组。感染后5 d分别口服2.00、1.00、0.50 g/mL的牛至中药组方汤剂,根据感染程度、存活率、每克粪便卵囊数(OPG)指标判断牛至组方汤剂对兔球虫病的治疗效果。结果表明,阳性对照组的临床症状比各用药组的临床症状严重,阳性对照组平均增重最少,中药组与阳性对照组相比差异极显著(P<0.01)。各试验组在感染后5 d均有卵囊排出,治疗组随着服药天数的延长OPG变为0,阳性对照组OPG明显高于治疗组。牛至中药组方中以高浓度组(2.00 g/mL)治疗兔球虫病效果最佳。说明牛至中药组方治疗兔球虫病有一定效果,且存一定的量效关系,该试验为牛至替代抗生素等药物预防和治疗兔球虫病提供理论依据。

基于accD基因序列的牛至遗传多样性研究

目的基于acc D基因序列分析不同产地牛至的遗传结构与分化,考察其遗传多样性。方法以8个居群69份样品为材料,试剂盒法提取植物基因组DNA,同源克隆法获得acc D序列,Sanger测序法双向测序,利用DNAsp、Arlequin等软件进行多态性分析,MEGA软件计算遗传距离并构建NJ发育树。结果牛至acc D长度为1432-1436 bp,存在10个单倍型与16个变异位点,单倍型多态性为0.595,核苷酸多样性为0.15750;牛至居群遗传分化较高(Gst=0.36751,Nst=0.46976,Fst=0.67627),基因流较低(Nm=0.43),居群间变异(67.62%)是主要的变异来源。系统发育关系显示10种单倍型形成3大分支,主要以H2为中心形成发散状的系统结构,种群未发生扩张。结论牛至居群存在较高的遗传多样性与遗传分化,可能主要与生境片段化、种群隔离及生活史特征相关,本研究为牛至种质资源的保护和开发利用提供理论基础。

牛至全日照间歇喷雾扦插育苗研究

[目的]研究牛至全日照间歇喷雾法扦插育苗的生根及成活情况。[方法]采用牛至不同插穗类型、不同基质配比进行扦插育苗,通过对生根及成活情况统计分析,研究不同因素对牛至全日照间歇喷雾法扦插育苗的影响。[结果]从生长点开始向下第一至第四节作插穗,插穗的发根率高,发根数量多,尤其第一节(生长点)的发根率、发根数量、根干重均表现最优。第一至第四节发根成活率分别为100%、100%、95.00%、92.50%,第五节及以下部位的插穗发根率及发根数量直线下降;对于不同基质,在砂:珍珠岩:营养土比例为1∶1∶1基质配比中第一至第四节发根成活率均为100%。[结论]在采用全日照间歇喷雾法进行牛至扦插育苗时,应选用的扦插基质砂:珍珠岩:营养土的比例为1∶1∶1;插穗选择从生长点开始向下第一至第四节发根成活率高,其中以第一节(生长点)的发根率、发根数量及成活率最优。

牛至挥发油化学成分鉴定及其主要成分对临床病原菌的抑制作用研究

探索牛至挥发油的化学成分及其主要成分对临床病原菌的抑制作用,筛选有效抑菌活性成分。通过GC-MS联用技术,分析牛至挥发油的化学成分及其含量;利用肉汤稀释法检测牛至挥发油对临床病原菌金黄色葡萄球菌、大肠杆菌、生物被膜介导的产ESBLs耐药大肠杆菌的抑制作用;利用96微孔板法研究牛至挥发油中主要成分1,8-桉树脑对耐药大肠杆菌生物被膜形成的影响。GC-MS联用技术鉴定出牛至挥发油中共有45种化学成分,占总挥发油的99.77%,主要成分是1,8-桉树脑(20.8%)、石竹烯(10.2%)、丁香油酚甲醚(9.8%)、香茅醇乙酸酯(8.8%)。牛至挥发油对临床病原菌金黄色葡萄球菌、大肠杆菌具有一定的抑制作用,其主要成分1,8-桉树脑可显著影响耐药大肠杆菌生物被膜的形成。牛至挥发油鉴定出45种化学成分,并具有较好的抑菌活性,其中1,8-桉树脑可作为潜在的生物被膜介导的耐药菌的耐药抑制活性成分。

牛至的化学成分研究(Ⅰ)

目的对牛至的化学成分进行分离和鉴定。方法应用溶剂法进行提取，硅胶、Sephadex LH-20及Toyopearl HW-40反复柱色谱分离、纯化，利用MS和NMR技术确定结构。结果从牛至氯仿部位分离鉴定了15个化合物，分别为β-谷甾醇（β-sitosterol，Ⅰ）、胡萝卜苷（dancosterol，Ⅱ）、二十六烯（hexacosene，Ⅲ）、二十六烷醇（hexacosanol，Ⅳ）、对二苯酚（1，4-benzenediol，Ⅴ）、邻二苯酚（1，2-benzenediol，Ⅵ）、对羟基苯甲醛（P-hydroxybenzaldehyde，Ⅶ）、4-甲基-5-异丙基邻苯二酚（thymgquinol，4-methyl-5-isopropyl-1，2-benzenediol，Ⅷ）、二氢脱氢二松柏醇（dihydrodehydrodiconiferyl alcohol，Ⅸ）、（一）-丁香脂酚[（一）-syringaresinol，Ⅹ]、琥珀酸（succinate acid，Ⅺ）、齐墩果酸（oleanolic acid，Ⅻ）、乌苏酸（usolic acid，ⅩⅢ）、咖啡酸乙酯（ethyl caffeate，ⅩⅣ）、迷迭香酸乙酯（ethyl rosmarinate，ⅩⅤ）。结论首次从牛至植物中分离得到苯丙素类化合物ⅠⅩ、Ⅹ，化合物Ⅲ～Ⅷ、Ⅺ、ⅩⅣ、ⅩⅤ亦为首次从该植物中分离得到。

牛至化学成分的研究

从药用植物牛至 ( Origanum vulgare L.)中分离并鉴定了 8个化合物 ;乌素酸( 1)、齐墩果酸 ( 2 )、原儿茶酸 ( 3)、日本椴甙 ( 4 )、Sagittatoside A( 5)、胡萝卜甙 ( 6)、β-谷甾醇 ( 7)、豆甾醇 ( 8)。2～ 5为首次从该种植物中分离得到。

牛至挥发油对小鼠特异性免疫功能的影响

The experimental results showed that volatile oil of Origanum vulgare L. (VO) significantly inhibited skindelayed type hypersensitivity (DTH) of mice induced by Dinitrochlorobenzene (DNCB) and sheep red blood cell(SRBC), and also inhibited the production of hemolysin in serum and SRBC hemolysis mediated by spleen cells atthe same dose of 100 mg·kg-1×7 d ip. The results also indicated it could cause thymus and spleen atrophyin mice at the dose. When the dose of VO was reduced to 50 mg·kg-1·d-1×7 d ip, it significantly increasedthe level of hemolysin in the serum of the mice and SRBC hemolysis mediated by spleen cells. VO couldremarkably inhibited DTH in mice induced by DNCB and SRBC at the dose. It suggested that VO could inhibit specificCell-mediated immunity and had different effects on specific humoral immunity with different doses.

3种不同方法提取牛至挥发精油及体外抑菌作用研究

采用水蒸气蒸馏法、同时蒸馏萃取法和微波辅助水蒸气法提取牛至挥发精油,并用气质联用分离鉴定,比较不同提取方法精油的得率及所得精油中具有抑菌功能的挥发性成分组成差异。将所得精油进行体外抑菌试验,比较3种方法所得精油对于供试菌的最小抑菌浓度。结果显示3种方法均得到不同含量的醇、烃、酸、醛、酮、酯和酚类物质,其主要具有抑菌功能的挥发性物质为石竹烯、β-水芹烯、蒈烯和柠檬烯等。

土香薷挥发油化学成分的研究

采用水蒸气蒸馏法从土香薷中提取挥发油,出油率为3.2％.利用GC-MS联用方法,在最佳分析条件下,共分离出51个峰,并对其化学成分进行了分析,鉴定出了45个化学成分,鉴定率为88.24％.其主要成分为:百里香酚(thymol)58.43％、香荆芥酚(carvacrol)19.29％、异己烷(isohexane)2.74％、甲基香荆醚(methyl carvacryiether)2.67％、乙酸牦牛儿酯(geranylacetane)0.96％、甲基百里醚(methylthymyiether)0.73％、对—聚伞花素(p-cymene)0.34％等.

白花茵陈挥发油主要化学成分及抑菌作用的研究

采用水蒸气蒸馏法从白花茵陈中提取挥发油,出油率为3.2%.利用GC-MS-DC联用方法,在最佳分析条件下,共分离出51个峰,并对其化学成分进行了分析,鉴定出了45个化学成分,鉴定率为88.24%.其主要成分为:百里香酚(thymol)58.43%、香荆芥酚(carvacrol)19.29%、异己烷(isohexane)2.74%、甲基香荆醚(methylcarvacryiether)2.67%、乙酸牦牛儿酯(geranylacetane)0.96%、甲基百里醚(methylthymyiether)0.73%、对-聚伞花素(p-cymene)0.34%等,并对白花茵陈挥发油的部分抑菌作用进行了研究.

气相色谱-质谱联用分析茵陈与牛至挥发油的化学成分

目的分析茵陈和牛至中挥发油成分,并比较其异同。方法按2010年版《中国药典》附录[挥发油测定法]中的规定,提取茵陈和牛至中的挥发油,用气相色谱-质谱法分析检测。结果茵陈和牛至挥发油的主要成分均为单萜类、双环倍半萜类成分。该两类成分在茵陈挥发油中质量分数分别为31.31%、68.69%;在牛至挥发油中质量分数分别为69.31%、20.25%。其中单萜类成分中两者共有成分的含量分别为17.89%、44.46%;双环倍半萜类成分中两者共有成分的含量分别为21.07%、14.0%。结论茵陈和牛至的挥发油中含有一些相同成分,但含量有较大差异。

致肠炎常见菌对牛至浸膏的敏感性试验

试验了３１株致肠炎常见菌对牛至浸膏的敏感性，结果表明，该浸膏对上述３１株菌均有不同程度的杀菌和抑菌作用。

牛至挥发油GC指纹图谱的研究

目的建立牛至挥发油的指纹图谱分析方法。方法采用气相色谱法,以DB-5石英毛细管柱为色谱柱,以高纯氮为载气,以二阶程序升温法,记录时间为80min,以氢火焰检测器检测,并以GC-MS对各共有峰进行了归属。结果用程序升温法得到了各色谱峰较好的分离,确定了22个共有峰。结论为更好地控制牛至药材的内在质量提供了可靠的方法。

牛至挥发油对肠炎常见菌的体外抗菌作用

不同浓度牛至挥发油对常见的致肠炎沙门氏菌属、大肠杆菌属和葡萄球菌属等31株菌的抗菌作用研究结果表明，该挥发油对上述31株菌均有不同程度的杀菌和抑菌作用．

药用植物牛至的组织培养技术研究

讨论了以牛至种子为外植体的组织培养繁殖技术。在增殖培养基的筛选过程中发现 ,以茎尖为外植体时 ,最适宜的增殖培养基为MS中加入BA 2 .0mg·L-1,IBA 0 .0 2mg·L-1,增殖倍数最高可达 5 .5 4 ;而茎段的增殖倍数较小为 2 .83。在生根培养基的筛选中发现 ,牛至在不含激素的 1/ 2MS、MS培养基上生根率可达 96 %～10 0 % ,但获得根较细 ,侧根少 ,移栽不易成活 ,加入激素后 ,生根数量增多 ,同时侧根数量明显增加 ,试验数据表明NAA ,IBA ,IAA 3种激素均可提高生根数量 ,浓度以 0 .5～ 1.0mg·L-1较好。