RESUMO
Leonurus japonicus Houtt. has been traditionally used to treat many ailments. This study evaluated the activating blood circulation, anti-inflammatory, and diuretic effects of L. japonicus extract (LJ) and identified its phytochemicals. In this work, the phytochemicals in LJ were identified using liquid chromatography mass spectrometry. Rats were randomly assigned to three groups (n=8): Control group was treated with saline, while the Model group (saline) and LJ group (426â mg/kg) had induced traumatic injury. All rats were treated with once by daily oral gavage for one week. The biochemical indices and protein expression were measured. Herein, 79 constituents were identified in LJ, which were effective in elevating body weight, food consumption, water intake, and urinary excretion volume, as well as in ameliorating traumatic muscle tissues in model rats. In addition, LJ prominently decreased the contents of plasma viscosity, platelet aggregation rate, thrombin time, prothrombin time, activated partial thromboplastin time, fibrinogen, thromboxane B2 (TXB2), TXB2/6-keto-prostaglandin F1α (6-keto-PGF1α), urokinase-type plasminogen activator (u-PA), plasminogen activator inhibitor 1 (PAI-1), PAI-1/tissue-type PA (t-PA), and PAI-1/u-PA, while significantly increasing antithrombin III, 6-keto-PGF1α, and t-PA contents. Furthermore, LJ notably inhibited tumor necrosis factor alpha, interleukin 6 (IL-6), IL-8, angiotensin II, antidiuretic hormone, aldosterone, aquaporin 1 (AQP1), AQP2, and AQP3 levels, and markedly elevating IL-10 and natriuretic peptide levels. Finally, LJ markedly reduced the protein expression of AQP1, AQP2, and AQP3 compared to the model group. Collectively, LJ possessed prominent activating blood circulation, anti-inflammatory, and diuretic effects, thus supporting the clinical application of L. japonicus.
Assuntos
Medicamentos de Ervas Chinesas , Hemostasia , Leonurus , Animais , Ratos , Anti-Inflamatórios , Aquaporina 2 , Diuréticos/farmacologia , Medicamentos de Ervas Chinesas/farmacologia , Leonurus/química , Leonurus/metabolismo , Inibidor 1 de Ativador de Plasminogênio/metabolismo , Ratos Sprague-Dawley , Ativador de Plasminogênio Tecidual/metabolismo , Ativador de Plasminogênio Tipo Uroquinase/metabolismo , Hemostasia/efeitos dos fármacos , Compostos Fitoquímicos/química , Compostos Fitoquímicos/farmacologiaRESUMO
Spiro-9,13-epoxy-labdane diterpenoids are commonly found in Leonurus species, particularly in Leonurus japonicus Houtt., which is a medicinal herb of long-standing use in Asia and in which such spiro-heterocycles are present in at least 38 diterpenoids. Here, through generation of a transcriptome and functional characterization of six diterpene synthases (diTPSs) from L. japonicus, including three class II diTPSs (LjTPS1, LjTPS3, and LjTPS4) and three class I diTPSs (LjTPS5, LjTPS6, and LjTPS7), formation of the spiro-9,13-epoxy-labdane backbone was elucidated, along with identification of the relevant diTPSs for production of other labdane-related diterpenes. Similar to what has been found with diTPSs from other plant species, while LjTPS3 specifically produces the carbon-9 (C9) hydroxylated bicycle peregrinol diphosphate (PPP), the subsequently acting LjTPS6 yields a mixture of four products, largely labda-13(16),14-dien-9-ol, but with substantial amounts of viteagnusin D and the C13-S/R epimers of 9,13-epoxy-labda-14-ene. Notably, structure-function analysis identified a critical residue in LjTPS6 (I420) in which single site mutations enable specific production of the 13S epimer. Indeed, extensive mutagenesis demonstrated that LjTPS6:I420G reacts with PPP to both specifically and efficiently produce 9,13S-epoxy-labda-14-ene, providing a specialized synthase for further investigation of derived diterpenoid biosynthesis. The results reported here provide a strong foundation for future studies of the intriguing spiro-9,13-epoxy-labdane diterpenoid metabolism found in L. japonicus.
Assuntos
Alquil e Aril Transferases , Diterpenos , Leonurus , Plantas Medicinais , Alquil e Aril Transferases/genética , Alquil e Aril Transferases/metabolismo , Diterpenos/metabolismo , Leonurus/metabolismo , TranscriptomaRESUMO
An HPLC quadrupole time-of-flight (Q-TOF) MS method was developed for the identification of secondary metabolites in Leonurus turkestanicus V.I. Krecz. et Kuprian. Ethanolic and chloroform extracts from the plant's aerial parts were tested. A total of 16 compounds (iridoid glycosides, phenylpropanoids, flavonoids, and nitrogen-containing compounds, as well as diterpene acetate derivatives) were identified and tentatively characterized based on (or using) their retention times and UV and Q-TOF-MS data. Previously reported aucubin (1), 6-deoxy-8-acetylharpagid (2), and stachydrine (13, 15) and homostachydrine isomers (14, 16) were identified, along with a lavandulifolioside isomer (3), verbascoside (4), rutin (5), 3-O-kaempferol rutinoside (6), and an unknown diterpene acetate (8). Compounds 3-6 were detected for the first time in this plant. Additionally. antimicrobial activity was evaluated. No significant differences were found between ethanolic and chloroform extracts of L. turkestanicus; however, the alcoholic extract showed stronger antifungal activity [minimal inhibitory concentration (MIC) of 2.5-5 mg/mL], whereas the chloroform extract showed stronger activity against the tested spore-forming Bacillus species (MIC 1.25-2.5 mg/mL).
Assuntos
Anti-Infecciosos/química , Anti-Infecciosos/farmacologia , Leonurus/química , Leonurus/metabolismo , Cromatografia Líquida de Alta Pressão , Avaliação Pré-Clínica de Medicamentos/métodos , Cazaquistão , Espectrometria de Massas/métodos , Metaboloma , Testes de Sensibilidade Microbiana , Componentes Aéreos da Planta/química , Extratos Vegetais/química , Plantas Medicinais/química , Metabolismo SecundárioRESUMO
In Leonurus sibiricus herb extract (LHE)-supplemented animals, plasma cholesterol decreased and high-density lipoprotein-cholesterol increased, resulting in a lowered atherogenic index. The plasma trolox equivalent antioxidant capacity, levels of hepatic thiobarbituric acid-reactive substances, and protein carbonyl values decreased significantly in LHE-supplemented mice (p<0.05), whereas the hepatic antioxidant indicators were all significantly elevated (p<0.05). In human umbilical vein endothelial cells stimulated with tumor necrosis factor alpha, LHE significantly suppressed intracellular reactive oxygen species, LOX-1, and adhesion molecules. LHE supplementation may modulate the lipoprotein composition and attenuate oxidative stress by elevated antioxidant processes, thus suppressing the activation of inflammatory mediators. This is a possible mechanism of the anti-atherogenic effect.
Assuntos
Moléculas de Adesão Celular/metabolismo , Células Endoteliais/efeitos dos fármacos , Hipercolesterolemia/metabolismo , Leonurus/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Extratos Vegetais/farmacologia , Receptores de LDL Oxidado/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Animais , Células Cultivadas , Feminino , Humanos , Lectinas/metabolismo , Camundongos , Veias Umbilicais/citologiaRESUMO
Motherwort (MW), a Korean folk medicine, has been applied to treat inflammatory disease. However, its effect on inflammatory cytokine release from mast cells is not well known. We investigated the anti- inflammatory effect of MW on the secretion of inflammatory cytokine such as tumor necrosis factor (TNF)-alpha and interleukin (IL)-6 and IL-8 in human mast cell line (HMC-1). MW was treated in vitro before activation of HMC-1 cells with phorbol 12-myristate 13-acetate (PMA) plus calcium ionophore A23187. MW had no cytotoxic effects on HMC-1 cell viability. MW (1 mg/ml) inhibited PMA plus A23187-stimulated gene expression and production of TNF-alpha, IL-6, and IL-8. Stimulation with PMA plus A23187 induced NF-kappaB activation in HMC-1 cells, which was inhibited by MW (1 mg/ml). MW inhibited secretion of TNF-alpha, IL-6, and IL-8 possibly by inhibiting NF-kappaB activation. These results indicate that MW may be helpful in regulating inflammatory diseases.
Assuntos
Anti-Inflamatórios/farmacologia , Leonurus/metabolismo , Mastócitos/efeitos dos fármacos , Extratos Vegetais/farmacologia , Anti-Inflamatórios/metabolismo , Calcimicina/farmacologia , Humanos , Inflamação/tratamento farmacológico , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Mastócitos/imunologia , Medicina Tradicional , NF-kappa B/metabolismo , Fitoterapia , Extratos Vegetais/metabolismo , Acetato de Tetradecanoilforbol/farmacologia , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Medicinal plants are valuable resources of natural antimicrobial materials. A novel small protein with antimicrobial activities, designated LJAMP1, was purified from the seeds of a medicinal herb, motherwort (Leonurus japonicus Houtt). LJAMP1 is a heat-stable protein with a molecular mass of 7.8 kDa and a determined isoelectric point of 8.2. In vitro assays showed that LJAMP1 inhibits the growth of an array of fungi and bacteria. The hyphal growth inhibition by LJAMP1 was more evident against hyphomycete fungi, such as Alternaria alternata, Cercospora personata, and Aspergillus niger. The N-terminal amino acid sequence of LJAMP1 was determined, and its coding gene was consequently cloned by the rapid amplification of cDNA ends. The gene LJAMP1 has no intron and encodes a polypeptide of 95 amino acids, in which the first 27 residues was deduced as a signal peptide. The mature LJAMP1 shows relatively low identity to plant napin-like storage proteins. Northern blot assays revealed that LJAMP1 is expressed preferentially in seeds. Bioassays in transgenic tobacco demonstrated that that overexpression of LJAMP1 significantly enhanced the resistance of tobacco against not only the fungal pathogen A. alternata but also the bacterial pathogen Ralstonia solanacearum, while no visible alteration in plant growth and development was observed.
Assuntos
Anti-Infecciosos/metabolismo , Bactérias/efeitos dos fármacos , Fungos/efeitos dos fármacos , Leonurus/metabolismo , Nicotiana/metabolismo , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Sequência de Aminoácidos , Anti-Infecciosos/química , Bactérias/patogenicidade , Sequência de Bases , Fungos/patogenicidade , Leonurus/crescimento & desenvolvimento , Dados de Sequência Molecular , Doenças das Plantas/microbiologia , Proteínas de Plantas/química , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/microbiologia , Sementes/metabolismo , Análise de Sequência de DNA , Nicotiana/genética , Nicotiana/microbiologiaRESUMO
Segments of sterile germinating plants were used to derive the tissue culture. Tests of the optimal hormonal composition of the medium from the standpoint of the growth of cultures revealed that the highest growth in biomass was achieved in the medium with an addition of 2.4-D (0.1, 1.0 mg.l-1), NAA (10.0 mg.l-1), and a combination of IAA (1.0 mg.l-1) + K (1.0 mg.l-1). The difference between these values was statistically insignificant. Other tested concentrations of growth regulators showed statistically significantly lower increases. The effects of different types of growth regulators and their concentrations on the production of selected metabolites in the tissue cultures were also examined and compared with the results obtained from the extract of the drug. Of alkaloids, stachydrine was demonstrated both in the extract of the drug and the extracts from all tested calluses regardless of the type and concentration of the growth regulator. Of flavonoid substances, two stains were demonstrated in the extract from the drug as well as in the extracts from all tested calluses. Neither of them was identical with the employed standards of rutin and quercetin. The results of TLC of iridoid substances were different in the extract from the drug (12 stains) and in the extracts from calluses (4 stains--3 identical with the stains in the extract from the drug). As to saccharides, three of them--glucose, fructose, and sucrose--were identified in the extract of the drug as well as in the extracts from all calluses.
Assuntos
Técnicas de Cultura , Leonurus/citologia , Plantas Medicinais/citologia , Leonurus/metabolismo , Reguladores de Crescimento de Plantas/farmacologia , Estruturas Vegetais/citologia , Estruturas Vegetais/metabolismo , Plantas Medicinais/metabolismoRESUMO
OBJECTIVE: To trace the effects of various microfertilizers on the growth and accumulation of total alkaloid in Leonurus artemisia. METHOD: The total alkaloid was determined by spectrophotometry. RESULT: Microfertilizers of different concentrations could facilitate the growth of L. artemisia in various degrees and Mn-fertilizer of standard concentration and Cu-fertilizer of excessive concentration could help raise the output of the plant markedly by up to 43.6% and 55.2% respectively. Microfertilizers of different concentrations could improve the content of total alkaloid in L. artemisia: Mn-fertilizer of standard concentration and B-fertilizer of excessive concentration could enhance the contents of total alkaloid up to 2.15% and 2.09% respectively. Mn-fertilizer and Cu-fertilizer of standard concentration have been proved very useful in improving the resistibility and ability of the plant to survive summer safely, and Fe-fertilizer of standard concentration could bring the plant an obvious increase of output after surviving summer. CONCLUSION: Application of Mn-fertilizer of standard concentration is very important for cultivating L. artemisia of excellent quality and high output.