Anti-inflammatory activity and identification of the Verbena litoralis Kunth crude extract constituents

Verbena litoralis is a plant popularly known as "gervãozinho-do-campo" in Portuguese. It is traditionally used for stomach, liver and gallbladder problems, and as an anti-inflammatory and anthelmintic. The goal of this study was to determine the chemical composition of the crude extract obtained from the aerial parts of V. litoralis by Ultra High Performance Liquid chromatography coupled with Mass Spectrometry in Tandem (UHPLC-MS/MS); assess the anti-inflammatory activity of ethyl acetate fraction and of crude extract; and verify liver, kidney and pancreas damage. In this study, the chemical composition of the extract was identified via UHPLC/MS/MS, assessing the anti-inflammatory activity of the crude extract and the acetate fraction in an induction model of the granulomatous tissue, as well as given liver, kidney and pancreas damage markers. Chlorogenic acid, luteolin, caffeic acid, apigenin, p-coumaric acid, vanillic acid, ferulic acid and quercetin were quantified in the extract. After the seven-day treatment, the granuloma of the animals treated with the plant extract and fraction presented values very close to the positive control (nimesulide). The V. litoralis crude extract and ethyl acetate fraction show anti-inflammatory activity similar to the nimesulide without evidence of liver, kidney and pancreas damage, which attributes the plant's pharmacological action to the flavonoids found.

Chromatographic fingerprint and multi-components quantitative analysis of Verbena officinalis by HPLC-CAD / 中草药

Objective: To establish a high performance liquid chromatography-charged aerosol detector (HPLC-CAD) method in order to obtain fingerprint profile and achieve the amount of isomer, oleanolic acid, and ursolic acid from Verbena officinalis. Methods: The method was developed by a C30 (150 mm × 2.1 mm, 3 μm) column and a linear gradient elution. Acenitrile-0.2% acetic acid was used as mobile phase. The flow rate was 0.3 mL/min; The injection volume was 10 μL and the column temperature was maintained at 20 ℃. Detector was Corona Ultra CAD with 40 oC of nebulization temperature. Results: Seventeen common peaks were observed from the fingerprint profiles of five different area samples. The relative standard divisions (RSDs) of retention time were less than 0.35%. The relative peak areas of the common peaks were in the range of 3.3%-44.0%. The contents of oleanolic acid were ranging from 0.097% to 0.136%, the contents of ursolic acid were ranging from 0.257% to 0.478%, the total contents were ranging from 0.354% to 0.478%. Conclusion: The fingerprint and simultaneous determination of oleanolic acid and ursolic acid in V. officinalis is firstly studied by HPLC-CAD. This method is simple and with good repeatability, which can be used for the quality control of V. officinalis.

Apoptosis of HepG-2 cells induced by total flavonoids of Verbena officinalis L. and possible mechanism / 国际药学研究杂志

Objective To explore the apoptosis and mechanisms of HepG-2 cells induced by total flavonoids of Verbena offici?nalis L.(TFV). Methods HepG-2 cells were cultured with different concentrations of TFV. The apoptosis of HepG-2 cells was evalu?ated by flow cytometry and DNA ladder. Reactive oxygen species(ROS)and membrane potential were measured by flow cytometry. Ex?pression of Caspase-3,Caspase-8,Caspase-9,and P53 was analyzed by Western blotting. Results TFV 50,100 and 200 mg/L in?creased the apoptosis rate(P<0.01),increased ROS levels of HepG-2 cells(P<0.01),and decreased the mitochondria membrane potential(P<0.05,P<0.01). TFV(50,100 and 200 mg/L)increased the proteins′ expression of Caspase-3,Caspase-9 and P53 (P<0.05,P<0.01). Conclusion TFV may induce the apoptosis of HepG-2 cells via increasing ROS levels,decreasing mitochon?dria membrane potential,and up-regulateing the expression of Caspase-3,Caspase-9 and P53 protein in HepG-2 cells.

Apoptosis of human hepatocellular carcinomaHepG-2 cells induced by total flavonoids of Verbena officinalis L through targeting topoisomerase Ⅱ / 中国药理学通报

Aim To explore whether total flavonoids of Verbena officinalis L(TFV)can induce apoptosis of HepG-2 cells through targeting topoisomerase Ⅱ.Methods HepG-2 cells were cultured with TFV at 200,100,50 mg·L-1.Cell apoptosis was evaluated by TUNEL-DAPI double staining.TopⅡ activity was detected by supercoiled pBR322 DNA relaxation assay.The levels of the mRNA of TOP Ⅱα,TOP Ⅱβ were analyzed by real time PCR.Expression of Bcl-2,Bax,TOP Ⅱα,TOP Ⅱβ and caspase-3 was analyzed by Western blot.

Establishment of HPLC fingerprint of Verbenae officinalis and determination of multi-components / 中草药

Objective: To establish an HPLC fingerprint method of Verbenae officinalis L. and determine the contents of its main components. Methods: Many batches of Verbenae officinalis fingerprint and the contents of main components were determined by HPLC, and the similarity and cluster analysis of the fingerprints of each batch were compared and the contents of the main components were compared. Results: The results showed that the difference of the main content of medicinal materials and fingerprints were between each batch of Verbenae officinalis. Conclusion: The fingerprint and its main components determination method have been established to provide the scientific basis for the quality evaluation of Verbenae officinalis.

Effects of Total Glucosides of Verbena officinalis on Chronic Nonbacterial Prostatitis in Mice and Its An-ti-inflammatory and Analgesic Effects Study / 中国药房

OBJECTIVE:To study the effects of total glucosides of Verbena officinalis on chronic nonbacterial prostatitis (CNP)in mice and its anti-inflammatory and analgesic effects. METHODS:60 mice were randomized into normal group(normal saline),model group(normal saline),positive group(Qianliekang tablet 1.5 g/kg)and total glucosides of V. officinalis high-dose, medium-dose and low-dose groups(0.2,0.1,0.05 g/kg),with 10 mice in each group. CNP model was induced in those groups ex-cept for normal group;mice were given relevant medicine once a day for consecutive 21 days since 8th day of modeling. The white blood cell count,lecithin corpuscle density and pathological changes of prostate samples were observed. 50 mice were randomly di-vided into model group(normal saline),positive group(Aspirin tablet 0.3 g/kg or Dexamethasone acetate tablet 0.01 g/kg)and to-tal glucosides of V. officinalis high-dose,medium-dose and low-dose groups(0.2,0.1,0.05 g/kg);they were given relevant medi-cine intragastrically once a day for consecutive 7 days. The xylene-induced ear swelling test (Aspirin tablet as positive drug) and cotton-pellet granuloma test (Dexamethasone acetate tablet as positive drug) were conducted to determine the the degree of ear swelling and the net weight of granuloma. The acetic acid writhing test and hot plate test(Aspirin tablet as positive drug)were car-ried out to determine the latency period of writhing,the times of writhing within 20 min and pain threshold. RESULTS:Compared with model group,white blood cells count of prostate tissue were lowered while the density of lecithin corpuscles were increased significantly in medicine groups;the pathology of prostate tissue were improved significantly in positive group and total glucosides of V. officinalis high-dose and medium-dose groups,and mostly were 0 and Ⅰ grade. The degree of ear swelling and the net weight of granuloma were markedly reduced in positive group and total glucosides of V. officinalis high-dose and medium-dose groups;the times of writhing within 20 min decreased in medicine groups,and the latency period of writhing prolonged and pain threshold in-creased in positive group and total glucosides of V. officinalis high-dose group. All of the results showed statistical difference(P<0.05 or P<0.01). CONCLUSIONS:Total glucosides of V. officinalis can improve CNP in mice,and shows good anti-inflammatory and analgesic effects.

Composición de los aceites esenciales de especies de Verbena nativas de la provincia de Buenos Aires bajo cultivo / Composition of the essential oils of Verbena species native to the province of Buenos Aires under cultivation

Verbena gracilescens (Cham.) Herter, V. bonariensis L., V. intermedia Gillies y Hook., V. litoralis Kunth, V. montevidensis Spreng. y V. rigida Spreng son las especies nativas del género en la provincia de Buenos Aires. Excepto V. montevidensis, el resto presenta propiedades hepatoprotectoras, digestivas, antiinflamatorias, antidiarreicas y antimicrobianas, entre otras. Este trabajo tiene por objetivo estudiar la composición de los aceites esenciales producidos por estas especies bajo cultivo durante dos temporadas en parcelas experimentales del campo de la Universidad Nacional de Luján. Las semillas utilizadas para el cultivo fueron colectadas de poblaciones naturales. Los aceites esenciales fueron obtenidos a partir de las flores de cada una de las especies por hidrodestilación y analizadas por Cromatografía de Gases-Espectrometría de Masas (GC-EM). La composición química de los aceites esenciales de Verbena en cultivo fue similar a la observada en estudios previos en poblaciones naturales. Los compuestos mayoritarios fueron fitol, 6,10,14- trimetil-2-pentadecanona (pentadecanona) y oct-1-en-3-ol (octenol). Sin embargo, las esencias obtenidas en el primer año de cultivo mostraron un alto contenido de fitol y muy bajo de pentadecanona, contrariamente a lo observado en los aceites esenciales del segundo año de cultivo. La cantidad de componentes de la segunda temporada aumentó con respecto a la primera. Estos cambios en la composición de los aceites esenciales durante el cultivo bianual de las especies podría deberse a las condiciones fisiológicas de las plantas según su edad o a las condiciones climáticas.

La Verbena azul en el tratamiento del niño con salpullido / The blue Verbena to treat the child with rash

Introducción: la miliaria es una enfermedad inflamatoria de la piel, usualmente conocida como salpullido, es muy frecuente en niños pequeños sobre todo en los meses de verano. Objetivo: valorar la eficacia de la infusión de Verbena azul en el control de la miliaria común. Métodos: se realizó un ensayo clínico fase II, a partir de la preparación de una infusión de hojas frescas o secas molidas, que se vertieron en 12 litros de agua hirviente, se tapó y se separó del fuego después de añádirle la planta, se dejó reposar y se coló. Fue aplicada tibia en todo el cuerpo durante 21 días, en dos secciones, al levantarse, y en la tarde luego del baño diario. El estudio se realizó en 30 lactantes de raza blanca, que acudieron con esta patología a la consulta de Medicina Natural y Tradicional, en el consultorio 6 del poblado de Altagracia, en el periodo entre junio del 2004 y junio del 2005, los cuales pertenecían al Policlínico Comunitario Docente Carlos J. Finlay de la Ciudad de Camagüey. Los datos se recogieron mediante una encuesta, confeccionada de acuerdo con los objetivos planteados en la investigación. Resultados: el grupo más afectado fue el de 3 a 6 meses, todos los pacientes fueron blancos, no se encontraron diferencias significativas en cuanto al sexo. La variedad clínica más frecuente fue la cristalina, y los meses en que más se presentó fueron entre junio y agosto. Conclusiones: este tratamiento presentó una alta eficacia en el control de la sintomatología de la miliaria común; se curaron 9 de cada 10 pacientes

Introduction: miliaria is an skin inflammatory disease known as rash and is very frequent in small children mainly during summer months. Objective: assessing the efficacy of the infusion of blue Verbena in controlling common heat rash. Methods: a phase II clinical trial was conducted from the preparation of an infusion of fresh leaves or dry grinded poured out in 12 liters of boiling water, was covered and removed from fire after adding the plant, it was doughed and strained. It was applied warm in whole body for 21 days in t wo sessions, to stand up and at evening after daily bath. A study was conducted in 30 white infants presenting with this pathology who came to the Natural and Traditional Medicine Service in the No.6 consulting room of the Altagracia settlement from June, 2004 to June, 2005 of Carlos J. Finlay Teaching Community polyclinic of the Camagüey city. Data were collected by survey, designed according to the objectives proposed in research. Results: the more involved group was that aged 3-6 months, all patients were white; there were not significant differences as regards sex. The more frequent clinical variety was the crystalline one and more prevalent months were June and August. Conclusions: present treatment was highly effective in the control of symptomatology of the common miliaria; nine of ten patients were cured

Functional Assessment of Plant Extracts by Application of Novel Neutrophil Activity Measurement System / 日本補完代替医療学会誌

<b>Objective:</b> The overproduction of reactive oxygen species leads to oxidative stress, which is related to lifestyle-related disease and cancer. Although antioxidants are considered as one of the countermeasures to oxidative stress, it is necessary to develop the assessment methodology for the antioxidant capacity which is closer to the inner body conditions. In this study, we examined antioxidant actions of plant extracts by using newly-developed neutrophil activity measurement system.<br> <b>Method:</b> Lemon verbena, Green tea, Camellia japonica and Antiallerge^® were used as plant extracts, and were diluted to medium in wide-range concentrations. Each solution was added on modified Mebiol Gel^® (hydrogel), and the mixture of blood and luminol were set onto the hydrogel in each tube. The amount of reactive oxygen species were measured by luminol-dependent chemiluminescence, whereas the cell count in the hydrogel was quantified as migratory activity of neutrophils.<br> <b>Result:</b> Lemon verbena and Green tea significantly inhibited reactive oxygen species in a concentration-dependent manner, whereas Camellia japonica and Antiallerge^® did not. The migratory activity of neutrophils was not affected by those plant extracts. Also, damaged cells were not detected.<br> <b>Conclusion:</b> It was suggested that Lemon verbena and Green tea scavenged reactive oxygen species without causing cell death of neutrophils. This new method for measuring neutrophil activities can be applied not only for assessing the status of inflammation and oxidative stress but also as the screening system for predicting the effectiveness of antioxidant and anti-inflammatory substances.<br>

Active caspase-3 detection to evaluate apoptosis induced by Verbena officinalis essential oil and citral in chronic lymphocytic leukaemia cells

Verbena officinalis L., Verbenaceae, commonly known as vervain, is a plant widely used in medicine. Despite of its widespread use in different traditional practices, the mechanisms of pharmacological actions of the plant and its volatile oil are still unclear. We evaluated the pro-apoptotic activity of V. officinalis essential oil and of its main component, citral, on lymphocytes collected from ten patients with chronic lymphocytic leukaemia (CLL), a disease in which a faulty apoptotic mechanism is still retained one of the primary pathogenic events, by adding to treated mononuclear cells, annexin-V, propidium iodide, and CD19. Apoptosis was also evaluated using anti-active-caspase-3 monoclonal antibody after permeabilization of the cells. Both V. officinalis essential oil and citral were found able to induce apoptosis in CLL cells and to activate caspase-3, which is considered the way by means they active apoptosis in B neoplastic cells. This data further support evidences that indicate natural compounds as possible lead structure to develop new therapeutic agents for CLL.

Studies on Iridoid Glucosides from Verbena officinalis / 中草药

For the purpose to seek the active components in the aerial parts of Verbena officinalis L.with anti-early pregnancy activity, the methanolic extract of this drug was repeatedly chromatographed onsilica gel column. Three iridoid glucosides were isolated and identified on the basis of physical, chemicalevidences and spectroscopic analysis (UV, IR, 1HNMR, 13CNMR, 1H-1HCOSY, 1H-13CCOSY and DEPT)as verbenalin (Ⅰ), 3, 4-dihydroverbenalin (Ⅲ) and hastatoside (Ⅲ).Ⅱ is a new natural compound.