Preparation and in vivo Distribution of Essential Oil from Alpinia zerumbet Fructus Encapsulated Nanoemulsions / 中国实验方剂学杂志

ObjectiveTo prepare oral nanoemulsions encapsulating essential oil from Alpinia zerumbet fructus（EOFAZ） and to investigate its pro-absorption effect in vitro and distribution in vivo. MethodThe proteoglycan conjugate polysaccharides of vinegar-processed Bupleuri Radix-bovine serum albumin（VBCP-BSA） was prepared by Maillard reaction of VBCP and BSA. Taking VBCP-BSA as emulsifier， vitamin B12（VB12） as absorption enhancer， and medium chain triglycerides mixed with EOFAZ as oil phase， the nanoemulsions loaded with EOFAZ was prepared by high energy emulsification method. The particle size， particle size distribution， surface Zeta potential， EOFAZ content and appearance and morphology of the nanoemulsions were characterized， and fluorescein tracer method was used to investigate the absorption effect of fluorescein-labeled EOFAZ nanoemulsions in vitro and their distribution in vivo. ResultVBCP-BSA was formed by Maillard reaction for 48 h with high grafting rate. Using VBCP-BSA as emulsifier， the homogeneous pink nanoemulsions was prepared and denoted as EOFAZ@VBCP-BSA/VB12. The particle size of the nanoemulsions was less than 100 nm and the particle size distribution was uniform. The surface of the nanoemulsions was a weak negative charge， and the shape was spherical. The encapsulation rate of the nanoemulsions for EOFAZ was greater than 80%， which had a good absorption effect in vitro and could enhance liver accumulation after oral administration. ConclusionThe designed proteoglycan nanoemulsions can effectively load EOFAZ， promote oral absorption and enhance liver distribution， which can provide experimental basis for the development of oral EOFAZ liver protection preparations.

Improvement effect and mechanism of total flavonoids from Alpinia zerumbet on gastric mucosa injury induced by absolute ethanol / 中国药房

OBJECTIVE To explore the improvement effect and potential mechanism of total flavonoids from Alpinia zerumbet on gastric mucosa injury induced by absolute ethanol through microRNA-146a-5p （miR-146a-5p）. METHODS Using human gastric mucosa GES-1 cells as objects， the acute gastric ulcer model was established by absolute ethanol； based on the investigation of the effects of different concentrations of total flavonoids from A. zerumbet on cell activity and the selection of action concentration， the relative expression level of miR-146a-5p in GES-1 cells was detected， the protein expressions of tumor necrosis factor （TNF） receptor-associated factor 6（TRAF6）， nuclear factor-κB p65 （NF-κB p65） and TNF-α were detected， and the levels of interleukin- 1β （IL-1β）， IL-6 and prostaglandin E2 （PGE2） in cell supernatant were determined. The targeting relationship between miR-146a- 5p and TRAF6 was verified； the effects of overexpressed miR-146a-5p and TRAF6 knockdown on the levels of IL-1β， IL-6 and PEG2 in supernatant of model cells as well as the effects of miR-146a-5p knockdown on anti-gastric ulcer effect of total flavonoids from A. zerumbet were observed. RESULTS Compared with the blank group， the relative expression of miR-146a-5p in cells and the level of PGE2 in cell supernatant were decreased significantly in the model group （P＜0.01）， while the protein expressions of TRAF6， NF-κB p65 and TNF-α in cells and the levels of IL-1β and IL-6 in cell supernatant were increased significantly （P＜ 0.01）. Compared with the model group， the relative expression of miR-146a-5p in cells and the level of PGE2 in cell supernatant were increased significantly in model+A. zerumbet total flavonoids （60 mg/L） group （P＜0.01）， while the protein expressions of TRAF6， NF-κB p65 and TNF-α in cells and 82260767） the levels of IL-1β and IL-6 in cell supernatant were decreased significantly （P＜0.05 or P＜0.01）. There was a targeted relationship and a negative correlation between miR-146a-5p E-mail：3113836821@qq.com and TRAF6. After overexpression of miR-146a-5p or TRAF6 knockdown， the levels of IL-1β and IL-6 were decreased significantly in cell supernatant， while the level of PGE2 was increased significantly （P＜0.05）. After miR-146a-5p knockdown， the levels of IL-1β and IL-6 in cell supernatant and the protein expression of TRAF6 in cells administered with total flavonoids of A. zerumbet were increased significantly， while the level of PGE2 was decreased significantly （P＜0.05）. CONCLUSIONS Total flavonoids of A. zerumbet can improve the gastric mucosa injury induced by absolute ethanol. The mechanism may be related to up-regulating the expression of miR-146a-5p， inhibiting the expression of TRAF6， and further inhibiting the secretion of related inflammatory factors.

Chemical constituents, larvicidal activity and molluscicidal from fresh leaves of Alpinia zerumbet (Pers.) and Cymbopogon citratus (DC.) Stapf / Constituyentes químicos, actividad larvicida y molusquicida de hojas frescas de Alpinia zerumbet (Pers.) y Cymbopogon citratus (DC.) Stapf / Constituintes químicos, atividade larvicida e moluscicida de folhas frescas de Alpinia zerumbet (Pers.) e Cymbopogon citratus (DC.) Stapf

SUMMARY Introduction: This study evaluated the chemical characterization, larvicidal activity and molluscicidal activity in front of the snail transmitting schistosomes (Biomphalaria glabrata) of the essential oils of Alpinia zerumbet and Cymbopogon citratus (DC.) Stapf. The essential oils (EOs) were extracted by hydrodistillation, with chemical characterization through Gas Chromatography coupled to mass spectrometry (GC-MS). The physical-chemical parameters were determined according to the Brazilian Pharmacopoeia. The toxicity test followed the bioassay with Artemia salina Leach, the EOs approved in this assay followed to evaluate their biological properties. Methodology: For molluscicidal activity, the methodology recommended by the WHO was performed, and the LC50 of the EOs was performed for their action in the face of the snail obtained by the Reed&Muench method. Both EOs showed low toxicity, and thus were evaluated for the biological properties larvicidal and molluscicidal. Alpinia zerumbet EO showed molluscicidal activity with LC50 of40.63 mg-L-1 and Cymbopogon citratus EO 33.94 mg-L-1. Results: Both EOs showed larvicidal and molluscicidal potentials against the organisms tested, showing satisfactory results for their action. The results indicate that the evaluated EOs are composed of substances that promote and encourage their application due to their potential for molluscicidal and larvicidal biological activity.

RESUMEN Introducción: Este estudio evaluó la caracterización química, la actividad larvicida y la actividad molusquicida frente al caracol que transmite esquistosomas (Biomphalaria glabrata) de los aceites esenciales de Alpinia zerumbet y Cymbopogon citratus (DC.) Stapf. Los aceites esenciales (AE) fueron extraídos por hidrodestilación, con caracterización química a través de cromatografía de gases acoplada a espectrometría de masas (GC-MS). Los parámetros físico-químicos se determinaron de acuerdo con la Farmacopea Brasileña. La prueba de toxicidad siguió al bioensayo con Artemia salina Leach, los AE probados en este ensayo se evaluaron a continuación en sus propiedades biológicas. Metodología: Para la actividad molusquicida se empleó la metodología recomendada por la OMS, y la LC50 de las AE se realizó para su acción frente al caracol obtenido por el método Reed&Muench. Ambos AE mostraron baja toxicidad, y por lo tanto fueron evaluados para las propiedades biológicas larvicidas y molusquicidas. Alpinia zerumbet AE mostró actividad molusquicida con LC50 de 40,63 mg-L'1 y Cymbopogon citratus EO 33,94 mg-L-1. Resultados: Ambos AE mostraron potenciales larvicidas y molusquicidas contra los organismos probados, mostrando resultados satisfactorios para su acción. Los resultados indican que los AE evaluados están compuestos de sustancias que promueven y fomentan su aplicación debido a su potencial para la actividad biológica molusquicida y larvicida.

RESUMO Introdução: Este estudo avaliou a caracterização química, atividade larvicida e ativi-dade moluscicida contra o caramujo transmissor de esquistossomos (Biomphalaria glabrata) dos óleos essenciais de Alpinia zerumbet e Cymbopogon citratus (DC.) Stapf. Os óleos essenciais (AE) foram extraídos por hidrodestilação, com caracterização química por cromatografia gasosa acoplada a espectrometria de massas (CG-EM). Os parâmetros físico-químicos foram determinados de acordo com a Farmacopéia Brasileira. O teste de toxicidade seguiu o bioensaio com Artemia salina Leach, os EA testados neste teste foram então avaliados quanto às suas propriedades biológicas. Metodologia: Para a atividade moluscicida, foi utilizada a metodologia recomendada pela OMS, e o LC50 do EA foi realizado para sua ação contra o caramujo obtido pelo método de Reed & Muench. Ambos os AE apresentaram baixa toxicidade e, portanto, foram avaliados quanto às propriedades biológicas larvicidas e moluscicidas. Alpinia zerumbet AE apresentou atividade moluscicida com CL50 de 40,63 mg-L-1 e Cymbopogon citratus EO 33,94 mg-L-1. Resultados: Ambos os AE apresentaram potencial larvicida e moluscicida contra os organismos testados, apresentando resultados satisfatórios para sua ação. Os resultados indicam que os AE avaliados são compostos por substâncias que promovem e estimulam sua aplicação devido ao seu potencial de atividade biológica moluscicida e larvicida.

Study on the Protective Effect of Alpinia zerumbet Extract on Acute Gastric Ulcer Model Mice / 中国药房

OBJECTIVE：To explore the protective effect of Alpinia zerumbet extract on acute gastric ulcer model mice. METHODS：Totally 48 mice were collected and randomly divided into blank group ，model group ，positive group （Sanjiu weitai granules，20 mg/kg），A. zerumbet extract high-dose ，medium-dose and low-dose groups （2.34，1.17，0.59 g/kg，by crude drug ）， with 8 mice in each group. They were given normal saline or relevant medicine intragastrically ，once a day ，for consecutive 7 d. Then，except for blank group ，other groups were given disposable intragastric administration of absolute ethanol 0.1 mL/10 g to establish acute gastric ulcer model. Another 48 mice were collected and grouped with same method ，and then given relevant medicine for consecutive 15 d. From the 10th day of administration ，except for blank group ，other groups were given intragastric administration of aspirin （20 mg/kg） for consecutive 6 d to establish acute gastric ulcer model. In ethanol induction model experiment，the formation of gastric ulcer and the index of gastric ulcer were observed in each group ；HE staining was used to observe the pathological changes of gastric tissue ；the levels of gastrin （GAS）in serum ，and the levels of SOD ，MDA and NO in gastric tissue were determined by ELISA. In the aspirin induced model experiment ，the formation of gastric ulcer and the index of gastric ulcer were observed and measured by the same method ；the levels of GAS ，TNF-α，IL-1β in serum，and the levels of PGE2 and COX- 2 in gastric tissue were determined by ELISA. RESULTS ：Compared with model group ，obvious ulcer lesions were found in gastric tissue of rats in model group ；the levels of GAS ，TNF-α，IL-1β in serum，and MDA ，NO in gastric tissue were increased significantly ， while the levels of SOD ， PGE2， （No.QZYY-2019- COX-2 in gastric tissue were decreased si gnificantly（P＜0.05 063）. Compared with model group ，the degree of gastric lesions in administration groups was alleviated to some extent；gastric ulcer index was decreased to some extent ， mail：997845460@qq.com while the levels of the above indexes in serum and gastric tissue were improved in varying degrees ， with statistical 制。E-mail：672863283@qq.com significance in most indexes （P＜0.05 or P＜0.01）. CONCLUSIONS：A. zerumbet extract exerts its protective effect on absolute ethanol and a spirin-induced acute gastric ulcer model mice，the mechanism of which may be relieving gastric mucosal injury through inhibiting oxidative stress and inflammatory response.

Research Progress on Miao Medicine of Alpinia zerumbet / 中国实验方剂学杂志

Alpinia zerumbet is a commonly used drug for ethnic minorities in Guizhou province. It has the effects of warming and drying dampness，relieving pain and eliminating malaria，and treating chills，chest and abdomen，indigestion，vomiting and diarrhea，with a long history of nearly 200 years. The author reviewed Chinese and foreign literatures for the past 30 years，and reviewed the research progress of the chemical constituents and pharmacological effects of A. zerumbet in and abroad，in order to provide a theoretical basis for its medicinal value. With the development of instruments and technology，the chemical composition of A. zerumbet has attracted more and more attention. More than 200 compounds have been isolated and identified，including volatile oils，flavonoids，steroids，terpenoids and organic acids and many other chemical ingredients. Pharmacological studies have shown that A. zerumbet has many pharmacological activities，such as anti-oxidation，blood pressure lowering，antispasmodic analgesia and protection against endothelial cells. However，current studies on the chemical constituents only focus on volatile oil components，and only a few studies focus on other organic matter and polysaccharides and are not deep enough. In clinic，the use of A. zerumbet is more confusing. The roots，stems，leaves，flowers，fruits and seeds can be used in medicines，but the pharmacological effects of the various medicinal parts have not been clearly explained，which leads to confusion in clinical medication. In addition，the pharmacological mechanism is not clear，especially in the studies on traditional activities，such as analgesic，digestive and anti-ulcer. The studies only focus on the pharmacological activity，and with only a few studies on the mechanism of action. Besides，the existing studies are mainly in vitro activity experiments，and need further validation in clinical trials，so as to provide reference for further rational development and comprehensive utilization of medicinal resources.

Essential Oil from Alpinia zerumbet Rhizome Inhibits Macrophage-derived Foam Cell Formation Via Modulating Expression of CD36 and ABCA1 / 中国实验方剂学杂志

Objective::To clarify the inhibitory effect of essential oil from Alpinia zerumbet rhizome (EOFAZ) on oxidized low-density lipoprotein (ox-LDL)-induced transformation of macrophage into foam cell and explore its possible mechanism. Method::THP-1 monocyte was incubated with 100 μg·L-1 phorbol myristate acetate (PMA) to grow into macrophage, experiment was divided into 4 groups as follows, control group, model group (80 mg·L-1 ox-LDL), EOFAZ at low dose (80 mg·L-1 ox-LDL+ 4 μg·L-1 EOFAZ)and EOFAZ at high dose (80 g·L-1 ox-LDL+ 20 μg·L-1 EOFAZ). Mathye thiazolye telrazliurn (MTT) method was employed to examine the influence of EOFAZ on macrophage viability. Western blot was used to analyze the expression level of cluster of differentiation 36(CD36) and ATP-binding cassette transporter A1(ABCA1) protein in macrophage. Enzyme-linked immunosorbent assay (ELISA) was used to detect cholesteryl ester contents in macrophage. Oil red O staining was applied to determine the accumulation of lipids in macrophage. Result::EOFAZ showed non-toxic effect on macrophage. Compared to control group, macrophage in model group displayed higher level of cholesteryl ester and lipid droplet(P<0.01), as well as significant increasing of CD36 expression (P<0.01), but no effect on ABCA1 expression. EOFAZ notably reduced the contents of lipids and cholesteryl ester(P<0.01), down-regulated expression of CD36 and up-regulated expression of ABCA1 in macrophage in comparison with the model group(P<0.01), indicating that EOFAZ inhibited transformation of macrophage into foam cell. Conclusion::EOFAZ could inhibit ox-LDL-induced transformation of macrophage into foam cell, the underlying mechanism may involves its ability to increase CD36 expression and decrease ABCA1 expression in macrophage.

Inhibitory Effect of Essential Oil from Fructus Alpinia zerumbet on Endothelial-to-mesenchymal Transition Induced by High Glucose via Nrf2/Notch1 Signaling Pathway / 中国实验方剂学杂志

Objective:To investigate the inhibitory effect and the possible mechanism of essential oil from fructus Alpinia zerumbet （EOFAZ） on endothelial-to-mesenchymal transition （EndMT） induced by high glucose （HG）. Method:Human umbilical vein endothelial cells （HUVECs） was cultured in vitro to analyze the pharmacodynamic effects of EOFAZ on EndMT and oxidative stress damage induced by HG. The experiment was set the blank group， HG group （35 mmol·L-1）， EOFAZ low dose group （1 μg·L-1） and EOFAZ high dose group （4 μg·L-1）. After EOFAZ intervention for 2 h， HG was added to incubate for 72 h in order to establish EndMT cell model. Western blot was used to detect the protein expression of vimentin and platelet endothelial cell adhesion molecule （CD31）. Angiogenesis experiment was used to detect the ability of cell migration ability in order to analyze the effect of EOFAZ on EndMT. The changes of reactive oxygen species （ROS） levels were detected by 2'，7'-dichlorodihydrofluorescein diacetate （DCFH-DA） fluorescence probe and the contents of malondialdehyde （MDA）， superoxide dismutase （SOD） and catalase （CAT） in cells were detected by the kit method to analyze the effect of EOFAZ on oxidative stress. Western blot was used to detect the protein expression levels of nuclear transcription factor E2 related factor 2 （Nrf2） and Notch1. The overexpression of Nrf2 was achieved by adenovirus （AD） transfection and the mechanism of EOFAZ inhibiting EndMT was further analyzed. The experiment was set the blank group， HG group （35 mmol·L-1）， AD-Nrf2 group， EOFAZ group （4 μg·L-1）， AD-Nrf2+EOFAZ group （4 μg·L-1）. The cells were infected with recombinant adenovirus overexpression plasmid of Nrf2 gene for 6 h， then replaced with normal medium for 24 h. After EOFAZ intervention for 2 h， HG was added to co-incubate for 72 h to induce EndMT. Western blot was used to detect the protein expressions of Nrf2， CD31， vimentin， Notch1 and Snail. Result:Compared with the HG group， after treatment with EOFAZ， the protein expression of CD31 was significantly up-regulated （P<0.05）， the protein expression of vimentin was significantly down-regulated （P<0.01）， the ability of cell migration was decreased （P<0.01）， and the contents of ROS and MDA were decreased （P<0.05， P<0.01）， the levels of CAT and SOD were increased （P<0.01）. In addition， EOFAZ could significantly up-regulate the protein expression of antioxidant signal Nrf2 （P<0.01） and down-regulate the protein expression of Notch1 （P<0.01）. High expression of Nrf2 was achieved by stable AD transfection into HUVECs. The results of Western blot showed that， compared with the HG group， the protein expression levels of Nrf2 and CD31 in each treatment group were significantly increased （P<0.01）， while the protein expression levels of vimentin， Notch1 and Snail were down-regulated （P<0.01）. At the same time， compared with the AD-Nrf2 group， the AD-Nrf2+EOFAZ group could further up-regulate the protein expressions of Nrf2 and CD31 （P<0.05， P<0.01）， while decrease the protein expression levels of vimentin， Notch1 and Snail （P<0.01）. Conclusion:EOFAZ ameliorates oxidative stress injury of vascular endothelial cells induced by HG and inhibits EndMT， which is related to Nrf2/Notch1 signaling pathway.

Antibacterial activity and bioactive compounds of 50% hydroethanolic extract of Alpinia zerumbet (Pers.) B.L. Burtt and amp; R.M. Sm

To evaluate antibacterial activity and the bioactive compounds of 50% hydroethanolic extract of Alpinia zerumbet (A. zerumbet) rhizomes. Methods: Eight reference microbial strains including two Gram-positive bacteria [Staphylococcus aureus (ATCC 29213) and Enterococcus faecalis (ATCC 29212)] and six Gram-negative bacteria [Escherichia coli (ATCC 25922), Klebsiella pneumoniae (ATTC 700603), Proteus mirabilis (DMST 8212), Salmonella enterica subsp. enterica serovar Vellore. (ATCC 15611), Shigella flexneri (ATCC 12022) and Pseudomonas aeruginosa (ATCC 27853)], were used to test antimicrobial susceptibility by the broth microdilution method. Bioactive compounds were analyzed by using HPLC. Results: The minimum inhibitory concentration values of A. zerumbet extract were 8 mg/mL for Staphylococcus aureus, Escherichia coli and Shigella flexneri and 16 mg/mL for Enterococcus faecalis and the other four Gram-negative bacilli. HPLC chromatograms revealed that the A. zerumbet extract contained hydroxybenzoic acids, hydroxycinnamic acids and flavonoids. Conclusions: The constituents of A. zerumbet rhizomes could be a potential source of antibacterial compounds, warranting further study of A. zerumbet extract.

Antibacterial activity and bioactive compounds of 50％ hydroethanolic extract of Alpinia zerumbet (Pers.) B.L. Burtt & R.M. Sm

Objective: To evaluate antibacterial activity and the bioactive compounds of 50％ hydro-ethanolic extract of Alpinia zerumbet (A. zerumbet) rhizomes. Methods: Eight reference microbial strains including two Gram-positive bacteria [Staphylococcus aureus (ATCC 29213) and Enterococcus faecalis (ATCC 29212)] and six Gram-negative bacteria [Escherichia coli (ATCC 25922), Klebsiella pneumoniae (ATTC 700603), Proteus mirabilis (DMST 8212), Salmonella enterica subsp. enterica serovar Vellore. (ATCC 15611), Shigella flexneri (ATCC 12022) and Pseudomonas aeruginosa (ATCC 27853)], were used to test antimicrobial susceptibility by the broth microdilution method. Bioactive compounds were analyzed by using HPLC. Results: The minimum inhibitory concentration values of A. zerumbet extract were 8 mg/mL for Staphylococcus aureus, Escherichia coli and Shigella flexneri and 16 mg/mL for Enterococcus faecalis and the other four Gram-negative bacilli. HPLC chromatograms revealed that the A. zerumbet extract contained hydroxybenzoic acids, hydroxycinnamic acids and flavonoids. Conclusions: The constituents of A. zerumbet rhizomes could be a potential source of antibacterial compounds, warranting further study of A. zerumbet extract.

In vitro efficacy of essential oils with different concentrations of 1, 8-cineole against Rhipicephalus (Boophilus) microplus / Eficácia in vitro de óleos essenciais com diferentes concentrações de 1, 8-cineol contra Rhipicephalus (Boophilus) microplus

Abstract The aim of this study was to evaluate the acaricidal activity of essential oils from three species of plants with intermediary concentrations of 1,8-cineole against the tick species Rhipicephalus (Boophilus) microplus. For this purpose, five serial concentrations (100.0, 50.0, 25.0, 12.5, 6.2 mg/mL) of essential oils from Mesosphaerum suaveolens (L.) Kuntze, Ocimum gratissimum L. and Alpinia zerumbet (Pers.) B. L. Burtt & R. M. Sm. were used on larval packet and adult immersion tests. The essential oils were analysed by gas chromatography-mass spectrometry (GC/MS) and gas chromatography-flame ionization detection (GC-FID), being detected 35.8, 24.7 and 24.0% of 1.8-cineol in the oils of M. suaveolens, O. gratissimum and A. zerumbet, respectively. The lethal concentration (LC 50) of each oil for larvae and engorged females was calculated through Probit analysis. All essential oils showed high efficacy (≥ 95.0%) on engorged females at the 100.0 mg/mL concentration. In regards to larvae, O. gratissimum (LC 50 = 11.9 mg/mL) was the most potent, followed by the A. zerumbet (LC50 = 19.7 mg/mL) and the M. suaveolens (LC50 = 51.6 mg/mL) essential oils. These results show that other compounds interfere with 1,8-cineole action.

Resumo O objetivo deste estudo foi avaliar a atividade acaricida de óleos essenciais de três espécies de plantas com concentrações intermediárias de 1,8-cineol contra o carrapato Rhipicephalus (Boophilus) microplus . Dessa forma, cinco concentrações diferentes (100,0; 50,0; 25,0; 12,5; 6,2 mg/mL) de óleos essenciais de Mesosphaerum suaveolens (L.) Kuntze , Ocimum gratissimum L. e Alpinia zerumbet (Pers.) B. L. Burtt & R. M. Sm. foram avaliadas pelos testes de pacote de larvas e de imersão de adultos. Os óleos essenciais foram analisados pela cromatografia gasosa acoplada a espectometria de massa (GC/MS) e cromatografia gasosa acoplada a detector de ionização de chama (GC-FID), sendo detectados 35,8, 24,7 e 24,0% de 1,8-cineol nos óleos de M. suaveolens, O. gratissimum e A. zerumbet , respectivamente. A concentração letal (CL50) de cada óleo essencial para larvas e fêmeas ingurgitadas foi calculada por meio da análise de Probit. Todos os óleos essenciais na concentração de 100,0 mg/mL apresentaram elevada eficácia (≥ 95,0%) sobre fêmeas ingurgitadas. Com relação as larvas, o óleo essencial de O. gratissimum (CL50 = 11,9 mg/mL) foi o mais potente, seguido pelos óleos de A. zerumbet (LC50 = 19,7 mg/mL) e M. suaveolens (LC50 = 51,6 mg/mL). Estes resultados demonstram que outros compostos interferem na eficácia de 1,8-cineol.

Composição química e eficácia do óleo essencial e do extrato etanólico de Alpinia zerumbet sobre Staphylococcus aureus / Chemical composition and efficacy of essential oil and ethanol extract of Alpinia zerumbet on Staphylococcus aureus

O presente trabalho descreve a atividade antibacteriana do óleo essencial e do extrato etanólico das folhas de Alpinia zerumbet (colônia) sobre cepas de Staphylococcus aureus isoladas de vacas com mastite subclínica e cepas padrão ATCC 29213 e ATCC 25923, por meio do método de difusão em ágar. Foram utilizados 10 tratamentos contendo diferentes concentrações do óleo essencial e do extrato etanólico (100,0; 50,0; 25,0; 12,5 e 6,3 mg.mL-1) e o grupo controle (álcool etílico a 50% e Tween a 1%). Os constituintes majoritários do óleo essencial foram p -cimeno (32,72%), 1,8-cineol (24,05%) e 4-terpineol (20,23%), sendo esses determinados por cromatografia a gás acoplada a espectrometria de massas e cromatografia a gás com detector de ionização de chama (CG-EM/DIC). No extrato etanólico foi detectado o ácido elágico e três flavonoides: rutina, quercetina e campferol, por meio de cromatografia a líquido de alta eficiência acoplada a detector de arranjo de diodo (CLAE-DAD). Todas as cepas apresentaram sensibilidade aos tratamentos com óleo essencial e extrato etanólico. A melhor resposta foi obtida com o óleo essencial de A. zerumbet que, na concentração de 100 mg.mL-1 proporcionou inibição total do crescimento bacteriano. Esses resultados sugerem o potencial antibacteriano do óleo essencial e do extrato etanólico de A. zerumbet no controle da mastite bovina.(AU)

The present work describes the antibacterial activity of the essential oil and the ethanol extract from leaves of Alpinia zerumbet (colônia) on Staphylococcus aureus strains isolated from cows with subclinical mastitis and standard strains ATCC 29213 and ATCC 25923, using the agar diffusion method. Ten treatments containing different concentrations of essential oil and ethanol extract (100.0; 50.0; 25.0; 12.5 and 6.3 mg.mL-1) and the control group (50% ethyl alcohol and 1% Tween solutions) were used for antimicrobial testing. The major constituents of the essential oil were p -cimeno (32.72%), 1.8-cineol (24.05%) and 4-terpineol (20.23%), which were determined by gas chromatographymass spectrometry and gas chromatography - flame ionization detector (CG-MS/FID). Ellagic acid and three flavonoids (rutin, quercetin and campferol) were detected in the ethanol extract by means of high performance liquid chromatography-photodiode array detector (HPLC-PDA). All strains showed sensitivity to the treatments with essential oil and the ethanol extract. The best response was obtained with A. zerumbet essential oil at a 100 mg.mL-1, showing complete inhibition of bacterial growth. These results demonstrate the antibacterial potential of essential oil and ethanol extract of A. zerumbet in the control of bovine mastitis.(AU)

Investigation of Neuropharmacological Activity of Seeds of Alpinia Zerumbet.

The study is based on the investigation of the neuropharmacological and analgesic properties observed from the methanolic extract of the seeds of Alpinia zerumbet. Tail immersion method in mice has been used for the evaluation of the central pharmacological actions. Similarly acetic-acid induced writhing-test was used for the evaluation of the peripheral pharmacological properties. A significant rise in pain threshold is seen in a dose dependent manner with the methanolic extract of A. zerumbet at doses of 100, 200 and 400 mg/kg body weight with the tail immersion methods. The methanolic extract at 400 mg/kg dose possessed 73.12% writhing inhibition, (p <0.001) in acetic-acid induced writhing-test that could be compared to the standard, Diclofenac-Na (25 mg/kg) with 75.78% inhibition. Open-field and hole-cross tests have been conducted in mice for further investigation of the extract in support of its neuropharmacological actions, where dose-dependent suppression of exploratory and motor activities were observed in the tested models. Hence, the above results evidence the presence of CNS depressant and analgesic properties of the plant, A. zerumbet.

Estudo químico e da atividade biológica cardiovascular do óleo essencial de folhas de Alpinia zerumbet (Pers.) B.L.Burtt & R.M.Sm. em ratos / Phytochemistry and cardiovascular biological activity of the essential oil from leaves of Alpinia zerumbet (Pers.) B.L. Burtt & R.M.Sm. in rats

A espécie vegetal Alpinia zerumbet (Pers.) B.L.Burtt & R.M. Sm. é popularmente empregada para o tratamento de diversas enfermidades, entre elas a hipertensão. Avaliar a composição química, a atividade antihipertensiva e ação na hipertrofia cardíaca do óleo essencial das folhas de Alpinia zerumbet (OEAZ) em ratos foram os objetivos deste estudo. O OEAZ, obtido por hidrodestilação em aparelho Clevenger, teve sua composição química analisada em cromatografia gasosa acoplada à espectrometria de massas (CG-EM). Foram identificados 14 constituintes, sendo terpinen-4-ol (37,45 por cento) o majoritário, seguido pelos óxido de cariofileno (7,56 por cento), trans-hidrato de sabineno (6,61 por cento) e 1,8-cineol (4,02 por cento). A avaliação cardiovascular foi feita após o tratamento crônico de ratos espontaneamente hipertensos (SHR) e seus respectivos controles, ratos Wistar-Kyoto (WKY). Os dados hemodinâmicos revelaram redução da pressão arterial média (PAM) no grupo tratado (SHRP: 160 ± 7 mm Hg; p<0,01) em relação ao não tratado (SHR: 180 ± 5 mm Hg). A relação entre peso do ventrículo esquerdo e peso corporal (VE/PC) do SHRP (2,50 ± 0,03 mg g-1; p<0,01) mostrou-se inferior ao SHR (2,61 ± 0,01 mg g-1), confirmando a redução da hipertrofia cardíaca (HC). Os dados de PAM e VE/PC dos animais SHRP foram estatisticamente diferentes quando comparados com os ratos controle (WKY: 116 ± 2 mm Hg e WKYP: 119 ± 4 mm Hg; p<0,05; WKY: 2,15 ± 0,04 mg g-1 e WKYP: 2,17 ± 0,04 mg g-1 ; p<0,01), indicando não ter havido normalização dos mesmos. Conclui-se que o tratamento crônico com OEAZ foi capaz de determinar redução, mas não a normalização, da PAM e da HC de ratos SHR, provavelmente pela presença dos componentes terpinen-4-ol e 1,8-cineol. Estudos com doses maiores ou período de tratamento superior são necessários para avaliar a possibilidade de o OEAZ normalizar os parâmetros analisados (PAM e HC).

Alpinia zerumbet (Pers.) B.L. Burtt & R.M.Sm. is traditionally employed to treat several diseases such as hypertension. The aim of this study was to evaluate the chemical composition, the anti-hypertensive activity and the capacity to reduce cardiac hypertrophy of the essential oil of A. zerumbet leaves (EOAZ) in rats. EOAZ was obtained through hydrodistillation in Clevenger apparatus and its chemical composition was analyzed by gas chromatography-mass spectrometry (GC-MS). Several constituents (14) were identified, terpen-4-ol (37.45 percent) being the major component, followed by caryophyllene oxide (7.56 percent), trans-sabinene hydrate (6.61 percent) and 1,8-cineol (4.02 percent). The cardiovascular effect was investigated after chronic treatment with spontaneously hypertensive rats (SHR) and their respective controls, Wistar-Kyoto rats (WKY). The treated group showed a lower mean arterial pressure (MAP) (SHRP: 160 ± 7 mm Hg; p<0.01) than the untreated group (SHR: 180 ± 5 mm Hg). The ratio of left ventricle-to-body weight (LV/BW) for SHRP was lower (2.504 ± 0.03 mg g-1; p<0.01) than that for SHR (2.162 ± 0.01 mg g-1), confirming the cardiac hypertrophy (CH) reduction. There were significant differences in MAP and CH between SHRP animals and control rats (WKY: 116 ± 2 mm Hg and WKYP: 119 ± 4 mm Hg; p<0.05. WKY: 2.152 ± 0.04 mg g-1 and WKYP: 2.168 ± 0.04 mg g-1; p<0.01), indicating that these values were not normalized. Those data showed that the chronic treatment with EOAZ reduces MAP and CH in SHR probably due to the presence of the compounds terpinen-4-ol and 1,8-cineol. Studies with higher doses or longer treatment periods are necessary to evaluate whether EOAZ can reduce the analyzed parameters (MAP and CH) to normal values.

Alpinia zerumbet (Pers.) B. L. Burtt & R. M. Sm. (Zingiberaceae): levantamento de publicações nas áreas farmacológica e química para o período de 1987 a 2008 / Alpinia zerumbet (Pers.) B. L. Burtt & R. M. Sm. (Zingiberaceae): survey of publications in pharmacological and chemical areas from 1987 to 2008

Espécie muito encontrada no nordeste do Brasil, Alpinia zerumbet (Pers.) B. L. Burtt & R. M. Sm. é conhecida popularmente como colônia. Na medicina popular é utilizada como antihipertensiva, diurética e febrífuga. Alguns estudos com óleos essenciais mostram sua atividade antimicrobiana, hipotensiva e um leve efeito diurético. Visando contribuir para o maior conhecimento da espécie realizou-se uma revisão de publicações envolvendo estudos nas áreas farmacológica e química para o período de 1987 a 2008.

Normally found in the northeast of Brazil, Alpinia zerumbet (Pers.) B. L. Burtt. & R. M. Sm. is commonly known as "colônia". In folk medicine, it has been used as antihypertensive, diuretic and febrifuge. Some studies with essential oils have shown its antimicrobial and hypotensive activities, besides a slight diuretic effect. Aimed at improving the knowledge of such species, a survey of publications involving studies in pharmacological and chemical areas was conducted from 1987 to 2008.

Vasodilator activity of extracts of field Alpinia purpurata (Vieill) K: Schum and A. zerumbet (Pers.) Burtt et Smith cultured in vitro / Vasodilatador de extratos de Alpinia purpurata campo (Vieill) K. Schum e A. zerumbet (Pers.) Burtt et Smith cultivadas in vitro

Nowadays, the high blood pressure is one of the main causes of death and cardiovascular diseases. Vasodilator drugs are frequently used to treat arterial hypertension. Experiments were undertaken to determine whether hydroalcoholic extracts obtained from leaves of field-grown Alpinia purpurata and A. zerumbet cultured in vitro under different plant growth regulators induce a vasodilator effect on Wistar rat mesenteric vascular bed pre-contracted with norepinephrine. Plant extracts were able to induce a long-lasting endothelium-dependent vasodilation. Efficiency on activity of A. purpurata reached 87 percent at concentration of 60 μg. The extract of A. zerumbet maintained in medium containing IAA, induced the relaxation (17.4 percent) at 90 μg, as compared to the control (MS0) that showed a better vasodilator effect (60 percent). These results are in agreement with the quantification of phenolic compounds in the extracts, which were 50 percent lower for those plants cultured in IAA. A. purpurata was assayed for the first time in relation to its vasodilator activity. This paper showed a strong probability of correlation between the pharmacological activities of A. purpurata with their content in phenolic compounds.

Atualmente, a hipertensão arterial é uma das maiores causas de morte e de doenças cardiovasculares. Os vasodilatadores são freqüentemente utilizados no tratamento da hipertensão. Extratos hidroalcoólicos de Alpinia purpurata de campo e de A. zerumbet cultivada in vitro sob diferentes reguladores de crescimento vegetal foram ensaiados no leito mesentérico de ratos Wistar. Os extratos de A. purpurata e A. zerumbet produziram efeito vasodilatador com padrão de resposta dose-dependente de duração prolongada. Extratos da espécie A. purpurata tiveram efeito vasodilatador de 87 por cento na dose de 60 μg. O extrato obtido de folhas de A. zerumbet oriundas das culturas mantidas em meio contendo AIA (ácido indol acético) inibiu o relaxamento (17,4 por cento) na dose de 90 μg em relação ao controle (MS0), com o qual foi verificado melhor efeito vasodilatador (60 por cento). Estes resultados estão de acordo com a concentração de fenóis totais que foi 50 por cento menor para os extratos de plantas cultivadas in vitro em AIA. A espécie A. purpurata foi pela primeira vez ensaiada quanto à atividade vasodilatadora. Os resultados obtidos indicaram a presença de substâncias fenólicas provavelmente correlacionadas à ação terapêutica de A. purpurata.

Chemical composition of the fractions of leaf oil of Alpinia zerumbet (Pers.) B.L. Burtt & R.M. Sm. and antimicrobial activity / Composição química de frações do óleo essencial de folhas de Alpinia zerumbet (Pers.) B.L. Burtt & R.M. Sm. e atividade antimicrobiana

Leaf oil prepared by hydrodistillation of Alpinia zerumbet Pers.) B.L. Burtt & R.M. Sm. was analyzed by GC/FID and GC/MS to determine the major compounds and it was also evaluated for antimicrobial activity. The oil presented a high content of oxygenated monoterpenes (52.5 percent), terpinen-4-ol, 1,8 cineole and γ-terpinene as the major constituents. The antimicrobial activity of leaf oil was tested by drop diffusion and bioauthography methods. Through drop test, all bacteria and fungi tested were inhibited by leaf oil. Preparative TLC (thin-layer chromatography) plates were developed using the mobile phase hexane: ethyl acetate (70:20, v/v) and fractions of the leaf oil were separated in three zones, scraped, extracted from silica and identified by GC/MS. The bioauthography method permitted to verify pronounced inhibition of Cryptococcus neoformans by fractions F2 and F3, both rich in oxygenated monoterpenes. The fraction F2 comprised 1,8 cineole (9.6 percent), linalool (3.7 percent) and caryophyllene oxide (5.4 percent), while fraction F3 showed mainly terpinen-4-ol (43.6 percent) and an amount of 32.7 percent oxygenated sesquiterpenes. The obtained data may be used to suggest the constituents of A. zerumbet leaf oil involved in antimicrobial activity.

O óleo essencial de folhas de Alpinia zerumbet (Pers.) B.L. Burtt & R.M. Sm., extraído por hidrodestilação, foi analisado por CG/DIC e CG/EM a fim de se determinar sua composição e atividade antimicrobiana. O óleo apresentou um elevado teor de monoterpenos oxigenados (52,5 por cento): terpinen-4-ol, 1,8-cineol e γ-terpineno como principais constituintes. A atividade antimicrobiana do óleo foi analisada pelos testes de difusão em agar e ensaios de bioautografia. Através do teste de difusão em agar, todas as bactérias e fungos testados foram inibidos pelo óleo essencial. Placas preparativas de CCD (cromatografia em camada fina) foram eluídas em hexano: acetato de etila (70:20, v/v), e o óleo foi separado em três frações que foram raspadas, extraídas da sílica e identificadas por CG/EM. A análise bioautográfica permitiu detectar pronunciada inibição de Cryptococcus neoformans pelas frações F2 e F3, ambas ricas em monoterpenos oxigenados. A fração F2 apresentou 1,8 cineol (9,6 por cento), linalol (3,7 por cento) e óxido de cariofileno (5,4 por cento), enquanto a fração F3 mostrou principalmente terpinen-4-ol (43,6 por cento) e cerca de 32,7 por cento de sesquiterpenos oxigenados. Os dados obtidos indicam os componentes do óleo de A. zerumbet envolvidos na atividade antimicrobiana.

Analysis on volatile oils from different parts of Alpinia zerumbet by GC-MS / 中成药

AIM:To analyze the chemical constituents of essential oils from three different parts of Alpinia zerumbet.METHODS:The oils were extracted by steam distillation.The chemical compounds were separated and identified by GC-MS.The relative content of each component was determined by area normalization.RESULTS:Fifty-five,sixty-seven,and sixty-six compounds from seed pericarp and capsule of Alpinia zerumbet were identified respectively.The ratios of identification of total compounds were 99.86%,95.83% and 95.52%,respectively.CONCLUSION:There are some differences among three different parts of Alpinia zerumbet.It provides the research base for advanced research and development.