Bacterial diversity in high Andean grassland soils disturbed with Lepidium meyenii crops evaluated by metagenomics / Diversidade bacteriana em solos de prados andinos elevados perturbados por culturas de Lepidium meyenii avaliadas por metagenômica

Soil quality is usually determined by its physical-chemical characteristics without taking into account the bacterial communities that play a fundamental role in the chemical decomposition of plant nutrients. In this context, the objective of the study was to evaluate bacterial diversity in high Andean grassland soils disturbed with Lepidium meyenii cultivation under different gradients of use (first, second and third use) and crop development (pre-sowing, hypocotyl development and post-harvest). The sampling was carried out in the Bombón plateau in the central Andes of Peru, during the rainy and low water seasons, by the systematic method based on a specific pattern assigned in a geometric rectangular shape at a depth of 0 - 20 cm. The characterization of the bacterial communities was carried out through the metagenomic sequencing of the 16S rRNA. 376 families of bacteria were reported, of which it was determined that there was a significant change in bacterial composition and distribution in relation to use pressure. There were no major changes due to the development of Lepidium meyenii. The families most sensitive to use pressure and soil poverty indicators were Verrucomicrobiaceae, Acidobacteraceae and Aakkermansiaceae.

A qualidade do solo é normalmente determinada pelas suas características físico-químicas sem ter em conta as comunidades bacterianas que desempenham um papel fundamental na decomposição química dos nutrientes das plantas. Neste contexto, o objetivo do estudo foi avaliar a diversidade bacteriana em solos de prados andinos elevados perturbados pelo cultivo de Lepidium meyenii sob diferentes gradientes de utilização (primeira, segunda e terceira utilizações) e desenvolvimento das culturas (pré-semeadura, desenvolvimento do hipocótilo e pós colheita). A amostragem foi realizada no planalto de Bombón, nos Andes centrais do Peru, durante as estações das chuvas e das águas baixas, pelo método sistemático baseado num padrão específico atribuído em forma geométrica retangular a uma profundidade de 0 - 20 cm. A caracterização das comunidades bacterianas foi realizada através da sequenciação metagenômica do rRNA 16S. Foram relatadas 376 famílias de bactérias, das quais se verificou uma alteração significativa na composição e distribuição bacteriana em relação à pressão de utilização. Não se registaram grandes alterações devido ao desenvolvimento do Lepidium meyenii. As famílias mais sensíveis à utilização de indicadores de pressão e pobreza do solo foram as Verrucomicrobiaceae, Acidobacteraceae e Aakkermansiaceae.

Bacterial diversity in high Andean grassland soils disturbed with Lepidium meyenii crops evaluated by metagenomics

Abstract Soil quality is usually determined by its physical-chemical characteristics without taking into account the bacterial communities that play a fundamental role in the chemical decomposition of plant nutrients. In this context, the objective of the study was to evaluate bacterial diversity in high Andean grassland soils disturbed with Lepidium meyenii cultivation under different gradients of use (first, second and third use) and crop development (pre-sowing, hypocotyl development and post-harvest). The sampling was carried out in the Bombón plateau in the central Andes of Peru, during the rainy and low water seasons, by the systematic method based on a specific pattern assigned in a geometric rectangular shape at a depth of 0 - 20 cm. The characterization of the bacterial communities was carried out through the metagenomic sequencing of the 16S rRNA. 376 families of bacteria were reported, of which it was determined that there was a significant change in bacterial composition and distribution in relation to use pressure. There were no major changes due to the development of Lepidium meyenii. The families most sensitive to use pressure and soil poverty indicators were Verrucomicrobiaceae, Acidobacteraceae and Aakkermansiaceae.

Resumo A qualidade do solo é normalmente determinada pelas suas características físico-químicas sem ter em conta as comunidades bacterianas que desempenham um papel fundamental na decomposição química dos nutrientes das plantas. Neste contexto, o objetivo do estudo foi avaliar a diversidade bacteriana em solos de prados andinos elevados perturbados pelo cultivo de Lepidium meyenii sob diferentes gradientes de utilização (primeira, segunda e terceira utilizações) e desenvolvimento das culturas (pré-semeadura, desenvolvimento do hipocótilo e pós-colheita). A amostragem foi realizada no planalto de Bombón, nos Andes centrais do Peru, durante as estações das chuvas e das águas baixas, pelo método sistemático baseado num padrão específico atribuído em forma geométrica retangular a uma profundidade de 0 - 20 cm. A caracterização das comunidades bacterianas foi realizada através da sequenciação metagenômica do rRNA 16S. Foram relatadas 376 famílias de bactérias, das quais se verificou uma alteração significativa na composição e distribuição bacteriana em relação à pressão de utilização. Não se registaram grandes alterações devido ao desenvolvimento do Lepidium meyenii. As famílias mais sensíveis à utilização de indicadores de pressão e pobreza do solo foram as Verrucomicrobiaceae, Acidobacteraceae e Aakkermansiaceae.

Bacterial diversity in high Andean grassland soils disturbed with Lepidium meyenii crops evaluated by metagenomics / Diversidade bacteriana em solos de prados andinos elevados perturbados por culturas de Lepidium meyenii avaliadas por metagenômica

Soil quality is usually determined by its physical-chemical characteristics without taking into account the bacterial communities that play a fundamental role in the chemical decomposition of plant nutrients. In this context, the objective of the study was to evaluate bacterial diversity in high Andean grassland soils disturbed with Lepidium meyenii cultivation under different gradients of use (first, second and third use) and crop development (pre-sowing, hypocotyl development and post-harvest). The sampling was carried out in the Bombón plateau in the central Andes of Peru, during the rainy and low water seasons, by the systematic method based on a specific pattern assigned in a geometric rectangular shape at a depth of 0 - 20 cm. The characterization of the bacterial communities was carried out through the metagenomic sequencing of the 16S rRNA. 376 families of bacteria were reported, of which it was determined that there was a significant change in bacterial composition and distribution in relation to use pressure. There were no major changes due to the development of Lepidium meyenii. The families most sensitive to use pressure and soil poverty indicators were Verrucomicrobiaceae, Acidobacteraceae and Aakkermansiaceae.

A qualidade do solo é normalmente determinada pelas suas características físico-químicas sem ter em conta as comunidades bacterianas que desempenham um papel fundamental na decomposição química dos nutrientes das plantas. Neste contexto, o objetivo do estudo foi avaliar a diversidade bacteriana em solos de prados andinos elevados perturbados pelo cultivo de Lepidium meyenii sob diferentes gradientes de utilização (primeira, segunda e terceira utilizações) e desenvolvimento das culturas (pré-semeadura, desenvolvimento do hipocótilo e póscolheita). A amostragem foi realizada no planalto de Bombón, nos Andes centrais do Peru, durante as estações das chuvas e das águas baixas, pelo método sistemático baseado num padrão específico atribuído em forma geométrica retangular a uma profundidade de 0 - 20 cm. A caracterização das comunidades bacterianas foi realizada através da sequenciação metagenômica do rRNA 16S. Foram relatadas 376 famílias de bactérias, das quais se verificou uma alteração significativa na composição e distribuição bacteriana em relação à pressão de utilização. Não se registaram grandes alterações devido ao desenvolvimento do Lepidium meyenii. As famílias mais sensíveis à utilização de indicadores de pressão e pobreza do solo foram as Verrucomicrobiaceae, Acidobacteraceae e Aakkermansiaceae.

Actividad fotoprotectora de una crema con extracto acuoso liofilizado de Lepidium meyenii (MACA) frente a la irradiación ultravioleta en piel de ratones / Photoprotective activity of a cream containing lyophilized aqueous extract of Lepidium meyenii (MACA) against ultraviolet irradiation on mouse skin

RESUMEN Objetivos: Evaluar la actividad fotoprotectora de una crema con extracto acuoso liofilizado de maca (ELM) frente a la irradiación ultravioleta (UV) en la piel de ratones. Materiales y métodos: Se realizó un estudio experimental en 35ratones BALB/c. Los tratamientos fueron aplicados por vía tópica en el dorso de los animales y posteriormente irradiados con rayos ultravioleta B, para luego medir el grosor en micras (µm) de muestras histológicas de la piel de los ratones. Se asignaron siete grupos divididos en no irradiado: blanco (G1) e irradiados con luz UV: sin tratamiento (G2); con protector solar comercial con factor de protección solar (FPS) 30 (G3); crema (placebo) (G4); ELM al 15% en agua (G5); ELM al 5% en crema (G6); y ELM al 15% en crema (G7). Se determinó el FPS in vitro, mediante el método de Mansur. Se realizaron las lecturas de las absorbancias en un espectrofotómetro ultravioleta-visible (UV-VIS) y se determinaron los FPS para las siguientes formulaciones: ELM al 5% en crema, benzofenona-4 (BZF-4) y bloqueador solar comercial FPS 30. Resultados: El grosor de piel de ratón en micras (µm) fue de 27,28 en G2; 18,31 en G3; 27,33 en G4; 19,51 en G5 y 18,04 en G6; no hubo diferencia significativa entre el grupo no expuesto a la radiación (G1) y el grupo ELM al 15% en crema (G7), ambos presentaron los menores grosores (12,76 y 14,20 µm, respectivamente). El FPS de ELM al 15% en crema fue 5,480 ± 0,020. Conclusiones: La formulación con ELM en crema presentó actividad fotoprotectora frente a la irradiación UV, los alcaloides fueron los componentes fitoquímicos mayormente encontrados y la formulación fue compatible con el activo (ELM).

ABSTRACT Objectives: To evaluate the photoprotective activity of a cream with lyophilized aqueous extract of maca (LEM) against ultraviolet (UV) irradiation in the skin of mice. Materials and methods: An experimental study was carried out on 35 BALB/c mice. Treatment was applied topically on the dorsum of the animals, which were subsequently irradiated with ultraviolet B rays, and then we measured the thickness in microns (μm) of histological samples of the skin of the mice. Seven groups were assigned, divided into non-irradiated: Blank (G1) and irradiated with UV light: no treatment (G2); with commercial sunscreen with sun protection factor (SPF) 30 (G3); cream (placebo) (G4); LEM at 15% in water (G5); LEM cream at 5% (G6); and LEM cream at 15% (G7). In vitro SPF was determined using the Mansur method. Absorbance readings were taken in an ultraviolet- visible spectrophotometer (UV-VIS) and SPFs were determined for the following formulations: LEM cream at 5%, benzophenone-4 (BZF-4) and commercial sunscreen SPF 30. Results: Mouse skin thickness in microns (μm) was 27.28 in G2; 18.31 in G3; 27.33 in G4; 19.51 in G5 and 18.04 in G6. There was no significant difference between the group not exposed to radiation (G1) and the 15% LEM cream group (G7), both had the lowest thicknesses (12.76 and 14.20 μm, respectively). The SPF of LEM cream at 15% was 5.480 ± 0.020. Conclusions: The formulation with LEM cream showed photoprotective activity against UV irradiation, alkaloids were the phytochemical components mostly found and the formulation was compatible with the active principle (LEM).

Phytochemical, antioxidant investigations and fatty acid composition of Lepidium sativum seeds

Aim: The aim of the present study was to characterize Lepidium sativum seed for phytochemicals, fatty acid composition and antioxidant properties.Methodology: Extraction of phytochemicals in different solvents, estimation of phenolics and flavonoids, physio-chemical properties, antioxidant assay by DPPH method was carried out. Seed oil content was determined by modified cold percolation extraction method and TD-NMR, followed by fatty acid compositional analysis using GLC. Results: The perusal of data revealed that the total oil content varied from 21.36% to 24.38%, and the major fatty acids identified were linolenic acid (24.447%) followed by oleic acid (24.413%) and palmitic acid (12.278%) . Physio-chemical and phytochemical characteristics were estimated for useful functional properties and markers viz. alkaloids, sugar and proteins. L. sativum seeds exhibited high antioxidant potential (0.063mg ml-1) compared to standard compounds such as ascorbic acid, rutin and quercetin.

Utilization of Garden Cress Seeds (Lepidium sativum L.) as Natural Source of Protein and Dietary Fiber in Noodles

The objective of this study was to investigate the effect of garden cress (Lepidium sativum L.) seeds on chemicalcomposition, cooking properties of noodle and antioxidant properties. Garden cress (GC) was added to semolina flour atdifferent levels 5, 10, 15 % (w/w), respectively. The chemical composition, cooking properties and sensory values of thesesamples were studied. The results showed an increasing level of protein, fat, ash, and fiber of garden cress noodles (GCN)by increasing the level of addition of garden cress (GC); while, a decrease of moisture content was noticed. The cookingquality properties of GCN were improved also by an increased level of garden cress (GC). Garden cress is a good source ofphenolic acids, flavonoids compound, and unsaturated fatty acids and for these reasons, garden cress is considered as afunctional food for due to health and nutritional values and its high content of protein and dietary fiber.

Induction of apoptosis in leukemic cells by the alkaloid extract of garden cress (Lepidium sativum L.) / 中西医结合学报

OBJECTIVE@#Garden cress (Lepidium sativum L.) is an important herb in traditional medicine used to improve production of breast milk in women and semen in men. In the present research the authors evaluated its ability to destroy leukemic cancer (Jurkat E6-1) cells, using the alkaloid extract of this plant.@*METHODS@#Constituents of the alkaloid extract were analyzed by gas chromatography-mass spectrometry (GC-MS) and their cytotoxicity in leukemic cancer cells and healthy peripheral blood mononuclear cells (PBMCs) was assessed. Cell death via apoptosis was confirmed by DNA laddering, caspase-3 activity, annexin V-fluorescein isothiocyanate and mitochondrial toxicity assays. The specific course of gene activation in treated cells was determined through quantitative polymerase chain reaction (qPCR).@*RESULTS@#GC-MS analysis identified six alkaloids and proto-alkaloids, namely, benzyl isothiocyanate (1), 2-ethoxy-4H-3,1-benzoxazin-4-one (2), (4R)-2-(2-aminophenyl)-4-phenyloxazoline (3), 5-acetyl-1,2-dihydro-6-methyl-2-oxo-4-phenyl-3-pyridinecarbonitrile (4), benzo[b][1,8]-naphthyridin-5(10H)-one,2,4,7-trimethyl (5) and 1,4-diaminoanthraquinone (6), in the alkaloid extract of L. sativum. Of these, compound 1 was previously identified in the seeds of L. sativum. Exposure to the alkaloid extract caused death of Jurkat E6-1 cells, with median lethal concentration (LC) of 75.25 µg/mL. However, the alkaloid extract also showed a nontoxic and proliferative (1.6-fold) effect in healthy PBMCs. Further experiments performed with Jurkat cells at LC and sub-LC doses demonstrated DNA fragmentation, activation of caspase-3 and time-dependant phosphatidylserine translocation (apoptosis) from inner to outer cell membranes. Cell toxicity and assessment of adenosine triphosphate level, together with using qPCR to evaluate expression profile of major apoptosis genes, revealed that apoptosis may be induced by disruption in the mitochondrial outer membrane potential, through activation of extrinsic and intrinsic apoptosis pathways in Jurkat cells.@*CONCLUSION@#The ability of the alkaloid extract of L. sativum seeds to induce apoptosis indicates a potential pharmacological use in cancer chemotherapy. The separation of individual active compounds and further in-depth exploration of the molecular mechanism of apoptosis may lead to novel chemotherapeutic compounds in our future antineoplastic research.

An analysis of volatile components in Lepidium salivum by GLME/GC-MS combined with retention index / 中草药

Objective: To analyze the difference between the volatile components of Lepidium salivum and its seeds. Methods: Gas chromatography-mass spectrometry (GLME) was performed for sample pretreatment, and gas chromatography-mass spectrometry (GC-MS) combined with retention index were used to determine the kinds and relative content of volatile components in the whole grass and seeds of L. salivum. Results: The results obtained were as follows: a total of 90 and 92 peaks were detected in whole plant and in seeds, respectively. Among them, 84 and 71 kinds of volatile components were successfully identified. However, there were differences in the types and relative content of the volatile components between the whole plant and the seeds. The main components in the whole plant were neophytadiene (15.39%), benzyl isothiocyanate (14.03%), 2-methoxy-4-vinylphenol (9.01%), 1,2-epoxyoctadecane (6.24%), benzaldehyde (5.60%), lignoceric alcohol (3.43%), stearyl aldehyde (2.37%) and benzyl nitrile (2.12%). While the major components in seed were benzyl nitrile (49.6%), benzyl isothiocyanate (10.51%), 2-(3,4-dimethylphenoxy) acetic acid (9.73%), benzoic acid, 2-(dimethylamino)ethyl ester (4.66%), benzaldehyde (3.63%), benzeneacetamide (3.18%), furfural (1.73%), and benzeneacetic acid (1.26%). As much as 24 kinds of volatile components were identified in both the whole plant and seed, such as benzyl nitrile, benzyl isothiocyanate, benzaldehyde, furfural and 2-methoxy-4-vinylphenol. Conclusion: This experiment provides essential data for further research on the role of volatile ingredients in pharmacologic action.

Producción de Pseudomonas sp. LMTK32 en medio modificado para peletizar semillas de maca (Lepidium meyenii Walp.) / Pseudomonas sp. LMTK32 production in modified media for pelleting seeds of maca (Lepidium meyenii Walp.)

Se produjo biomasa de Pseudomonas sp. LMTK32 a partir de la modificación del medio de cultivo Caldo Extracto de Levadura Manitol (LMC) con el objetivo de incrementar el número de células viables con capacidad de promover la germinación de semillas de maca peletizadas y reducir los costos de producción. En el proceso de optimización, los componentes extracto de levadura y manitol del medio de cultivo LMC fueron reemplazados por fuentes comerciales de sacarosa y glutamato, cuyas concentraciones fueron determinadas en matraces mediante el diseño estadístico de Box-Behnken; además, se determinó el efecto del porcentaje de inóculo en el tiempo de producción de biomasa. Posteriormente se determinó a nivel de biorreactor que 28.57 h-1 fue el valor adecuado del coeficiente volumétrico de transferencia de oxigeno (kLa) a 600 rpm, produciendo 1.28x1011 UFC/mL. En el medio modificado M1, empleando 12.06 g/L-1 de sacarosa, 11.50 g/L-1 de glutamato de sodio y 10.9% de inoculante se obtuvo 15x108 UFC/mL, superando en 52% más el número de células viables con respecto al tratamiento control LMC (7.8x108 UFC/mL). A nivel in vitro, la peletización de semillas de maca con Pseudomonas sp. LMTK32 producidas en biorreactor y en el medio modificado M1 favoreció su germinación. A partir de sustratos orgánicos comerciales se puede producir inoculantes bacterianos eficientes en el desarrollo de cultivos de maca, sin alterar su capacidad de promover el crecimiento vegetal

Biomass of Pseudomonas sp. LMTK32 was produced from modification of culture media Yeast Extract Mannitol Broth (YEMB) with the aim of increasing the number of viable cells with the ability to promote the germination of maca seeds pelleted with the bacteria and reduce production costs. In the optimization process, the yeast extract and mannitol components of the LMC culture media were replaced by commercial sources of sucrose and glutamate, whose concentrations were determined in flasks by statistical design from Box-Behnken; in addition, the effect of the inoculum percentage on the time of biomass production was determined. Subsequently, it was determined at the bioreactor level that 28.57 h-1 was the adequate value of the volumetric oxygen transfer coefficient (kLa) at 600 rpm, producing 1.28 x 10 11 CFU / mL. In the LMC M1 modified media, using 12.06 g / L-1 of sucrose, 11.50 g / L-1 of sodium glutamate and 10.9% of inoculant obtained 15x108 CFU / mL, increasing in 48% the number of viable cells with respect to the YEMB control treatment (7.8x10 8 CFU / mL). At the in vitro level, the pelleting of maca seeds with Pseudomonas sp. LMTK32 produced in bioreactor and in the modified media M1 favored its germination. From commercial organic substrates, efficient bacterial inoculants can be produced in the development of maca crops, without altering their ability to promote plant growth

Research and application progress of Lepidium meyenii（maca） / 中国中药杂志

Lepidium meyenii（maca）was a herbaceous plant of the family Cruciferae. It is native to the andes region of South America where the local people had been growing and consuming maca for centuries. The unique chemical composition and physiological function of maca were widely concerned worldwide. It was introduced to China in 2002, and were cultivated successfully in Yunnan, Tibet, Sichuan, Jilin and other places with a certain size. Maca contained not only rich nutrition such as protein, vitamin and mineral matter, but also lots of secondary metabolites as maca alkaloids, glucosinolates, volatile oils, sterols polyphenols and macaenes. Numerous studies suggested that maca may serve effects in resisting oxidation, fatigue resistance, raising fertility, regulating endocrine, enhancing immunity, tumour suppression, treating osteoporosis, regulate blood sugar and protection of nervous system. Maca was approved by the Ministry of Health as a new resource food in 2011, and its related products include food, health foods, cosmetics, etc. Certain exploratory researches were carried to take better advantage of maca's medicinal value. This paper briefly reviewed the research and application progress of maca in recent years from the aspects of botany, chemical composition, function, resources situation and related products development, which was supposed to provide reference for scientific research and utilization of maca.

Lignans from seeds of Lepidium apetalum / 中草药

Objective To study the chemical constituents from the seeds of Lepidium apetalum. Methods Compounds were isolated from the water extract from the seeds of L. apetalum by using Diaion HP-20, Toyopearl HW-40, MCI Gel CHP-20, ODS, Silica gel chromatography and semi-preparative-HPLC. Results Nine compounds were isolated and identified as lepidiumlignan A (1), lepidiumlignan B (2), erythro-1-(4-O-β-D-glucopyranosyl-3-methoxyphenyl)-2-[4-(3-hydroxypropyl)-2,6-dimethoxyphenoxy]-1,3- propanediol (3), (7R,7’E,8S)-4,9-dihydroxy-3,3’,5-trimethoxy-4’,7-epoxy-8,5’-neolign-7’-en-9’-oic acid (4), spicatolignan B (5), (-)-pinoresinol-4-O-β-D-glucopyranoside (6), (-)-isolariciresinol (7), aegineoside (8), and (+)-syringaresinol-O-β-D-glucopyra- noside (9). Conclusion Compounds 1 and 2 are new compounds, named as lepidiumlignan A and lepidiumlignan B. Compounds 3-9 are isolated from the plants for the first time.

A new macamide from Lepidium meyenii cultivated in Lijiang / 中草药

Objective To study the chemical constituents of the dried roots of Lepidium meyenii cultivated in Lijiang. Methods The samples were extracted by 90% alcohol, and then isolated by silica column, MCI, and HPLC. The structures of the isolated compounds were elucidated by NMR techniques including 2D NMR such as HSQC, HMBC, and COSY. The cytotoxic activity of the new compound against five cell lines (NB4, A549, PC3, SHSY5Y, and MCF7) was evaluated by MTT methods. Results Three macamides were isolated and identified as N-(3,4-dimethoxybenzyl) hexadec-9Z-enamide (1), N-benzyl-13-oxo-9E,11E-octadecadienamide (2) and N-(3,4-dimethoxybenzyl)-hexadecanamide (3) from this plant. Conclusion Compound 1 is a new compound named macalepidiumide A and shows no significant cytotoxic activity.

Cloning, sequence analysis, and prokaryotic expression of LaF3H gene from Lepidium apetalum / 中草药

Objective In order to study the key genes involved in flavonoid biosynthesis pathway, the flavanone-3-hydroxylase (F3H) gene was isolated from Lepidium apetalum, which is named as LaF3H. Meanwhile, the sequence analysis, prokaryotic expression, and purification were also performed. Methods Specific primers were designed according to LaF3H gene sequences in the transcriptome data of L. apetalum, and the cDNA sequence of LaF3H gene was isolated from L. apetalum. By construction the prokaryotic expression vector pET-32a-LaF3H, the recombinant LaF3H protein was expressed in Escherichia coli BL21 (DE3) cells under IPTG induction. Results The open reading frame (ORF) of LaF3H was 1 080 bp, which encoded a protein of 359 amino acid residues, with a predicted molecular mass of 40 320. Sequence analysis showed that LaF3H contains five conserved motifs of F3H protein. The phylogenetic analysis indicated that LaF3H protein showed the highest homology with F3H protein from cruciferous plants (such as AtF3H from Arabidopsis thaliana). The prokaryotic expression vector pET-32a-LaF3H was constructed and the recombinant LaF3H protein was successfully expressed in E. coli BL21 (DE3) cells. Furthermore, the recombinant LaF3H protein was purified through Ni2+ affinity chromatography. Conclusion The LaF3H gene was isolated from L. apetalum and the recombinant LaF3H protein was obtained. The results of this study provided the foundation for the further preparation of LaF3H antibody and detection of LaF3H enzyme activity, and were helpful for functional characterization of LaF3H gene involved in flavonoid biosynthesis pathway of L. apetalum.

Ethanol extracts of Lepidium meyenii Walp (maca) from two different areas in Xinjiang promote the maturation of mouse macrophages and dendritic cells / 中华微生物学和免疫学杂志

Objective To investigate the effects of ethanol extracts of Lepidium meyenii Walp (LMEE) from two different areas in Xinjiang on the maturation of mouse macrophages (RAW264. 7 cells) and dendritic cells (DCs). Methods Ethanol extracts of LMEE from Tashikuergan County (Ta xian) and A La gou of Xinjiang were prepared and named as LMEE-T and LMEE-A, respectively. RAW264. 7 cells and bone marrow-derived DCs from C57BL/6 mice were treated with different concentrations of LMEE-T/A. The viability of RAW264. 7 cells was analyzed by MTT assay. Expression of costimulatory molecules and MHCⅠ on the surface of RAW264. 7 cells and DCs was detected by flow cytometry. Secretion of cytokines and the release of nitrogen oxide (NO) were measured by ELISA and Griess method, respectively. Results LMEE-T/A had no significant influence on the viability of RAW264. 7 cells when the concentration was lower than 1 mg/ml. Treating RAW264. 7 cells with LMEE-T/A promoted surface molecule expression, cytokine secretion and NO release through TLR4 signaling pathway in a dose-dependent manner. Moreover, LMEE-T was more potent than LMEE-A. LMEE-T/A at the concentration of 0. 4 mg/ml promoted the expression of DC surface molecules and the secretion of cytokines. Infrared and ultraviolet spectra showed that LMEE-A and LMEE-T contained polysaccharides, macaenes, macamides and flavanols. Compared with LMEE-A, LMEE-T contained more benzene ring compounds but less polysaccharides. Conclusion Both LMEE-T and LMEE-A could activate RAW264. 7 cells and promote the maturation of DCs. The differences between their effects might be related to the differences in their contents.

Effects of water extracts of Lepidium meyenii walp (maca) growing in two different places in Xin-jiang on the maturation and function of dendritic cells / 中华微生物学和免疫学杂志

Objective To investigate the effects of water extracts of Lepidium meyenii walp (LMWE) collected from two different places in Xinjiang on the maturation and function of dendritic cells (DCs) and to evaluate their roles in immunoregulation. Methods Water extracts of LMWE growing in Tashikuergan County (Ta xian) and A La gou of Xinjiang were prepared and named as LMWE-T and LMWE-A,respectively. Bone marrow-derived DCs and splenocytes isolated from C57BL/6 mice were treated with different concentrations of polysaccharide extracts from LMWE-T/A. Effects of LMWE-T/A on the per-centage and apoptosis of DC,expression of co-stimulatory molecules and secretion of cytokines were detected by flow cytometry and ELISA. MTT assay was used to analyze the proliferation of splenocytes. Changes in the functions of DC were evaluated by mixed lymphocyte reaction(MLR). Results LMWE-T/A had no in-fluence on the percentage and viability of DC. Expression of CD40 and CD86,and secretion of TNF-α,IL-12p40 and IFN-γ were significantly increased by LMWE-T/A treatment in a dose-dependent manner. LMWE-T/A treatment enhanced the functions of DCs and also dose-dependently promoted the proliferation of splenocytes. LMWE-A was more effective than LMWE-T in promoting CD86 expression,IFN-γ secretion and splenocyte proliferation. Pretreatment with TAK-242,an inhibitor of Toll-like receptor 4(TLR4),could sig-nificantly inhibit the regulatory effects of LMWE-T/A on CD40 expression and the secretion of TNF-α and IL-12p40. Conclusion This study shows that LMWE could promote the maturation of DC through TLR4 signaling pathway,enhance the functions of DC without side effects on DC survival,and increase the prolif-eration of splenocytes,indicating that LMWE has a potential immunopotentiating effect. LMWE-A has better effects than LMWE-T on immune enhancement.

Chemical Constituents from the Seeds of Lepidium apetalum Willd / 中国药学杂志

OBJECTIVE: To investigate the chemical constituents of the seeds of Lepidium apetalum Willd. METHODS: The compounds were isolated and purified by Diaion HP-20, Toyopearl HW-40, MCI Gel CHP-20, ODS, silica gel chromatography combined with Pre-HPLC and the structures were identified on the basis of spectral data and physiochemical properties. RESULTS: Sixteen compounds were isolated and identified from the water extract as catechol(1), protocatechuic aldehyde(2), 2-phenyl acetamide(3), methyl-5-hydroxypyridine-2-carboxlate(4), benzylcarbamic acid(5), N-benzylacetamide(6), raphanuside C(7), 1-phenyl-1, 2-ethanediol(8), 2-(4-hydroxyphenyl)-ethanol(9), isorhamnetin-7-O-α-L-rhamnopyranoside(10), kaempferol(11), methyl 2, 4, 6-trihydroxybenzoate(12), 2-(4-hydroxyphenyl)acetonitrile(13), syringic acid(14), protocatechuic acid(15), and methyl sinapate(16). CONCLUSION: Compounds 1-16 are isolated from this plant for the first time.

Protective effect of Lepidium sativum seed extract on histopathology and morphology of epididymis in diabetic rat model / Efecto protector de extracto de semilla Lepidium sativum sobre la histopatología y morfología del epidídimo en modelo de rata diabética

Diabetes mellitus is a frequent and serious metabolic illness all over the world and plants have been a desirable source of medicine recently. Diabetes has unpleasant effect on male reproductive system and it may lead to male infertility. It causes erectile dysfunction and reduces ejaculate volume by affecting the health of small blood vessels and the small nerves that control ejaculation and also decreases libido by decreasing testosterone levels. Current study evaluated the possible protective efficiency of Lepidium sativum (Garden cress) seed extract on fasting blood sugar (FBS) and then assessed histopathological change of epididymis in streptozotocine (STZ)-induced diabetic rats. We randomly categorized 50 adult male Wistar rats into five groups (each 10 rats). Group 1 was control placebo group receiving only 0.1 ml normal saline via gastric gavages, Group 2 as control diabetic rats received an intraperitoneal (IP) injection of STZ 60 mg/kg body weight. Rats with FBS >250 mg/dl were considered as diabetic. Group 3 were diabetic rats receiving insulin in dose 3U/100 g body weight and Groups 4 and 5 were diabetic rats that received 0.1 cc of 200 and 400 mg/kg, ethanol extract of Lepidium sativum seed by gavages daily. One day after the last gavages, rats were anesthetized by chloroform. Epididymis duct was removed from abdomen and weighed with a digital scale. Afterwards, samples were putted in Bouin's solution for histological measurement. Administration of 200 and 400 mg/ml doses of Lepidium sativum seed extract increased epithelium height and decreased interstitial volume density and fibro muscular thickness significantly. Also, volume density of epithelium, fibro muscular, lumen and interstitial decreased significantly. Tubular and lumen diameter did not change significantly in different groups. It appears Lepidium sativum seed extract is a beneficial protective supplementary agent against adverse effects of diabetes on male reproductive system.

La diabetes mellitus es una enfermedad metabólica frecuente y grave que afecta a los hombres en todo el mundo. Recientemente, las plantas han sido una fuente deseable de medicina para este tipo de enfermedad. La diabetes tiene un efecto perjudicial en el sistema reproductivo masculino y puede conducir a la infertilidad. Causa disfunción eréctil y reduce el volumen de la eyaculación al afectar los pequeños vasos sanguíneos y los nervios que controlan la eyaculación. También disminuye la libido reduciendo los niveles de testosterona. El presente estudio evaluó la posible eficacia protectora del extracto de semilla de Lepidium sativum en la glucemia en ayunas y también se evaluó el cambio histopatológico del epidídimo en ratas diabéticas inducidas por estreptozotocina (STZ). Se dividieron aleatoriamente 50 ratas Wistar macho adultas en cinco grupos de 10 ratas cada uno. El grupo 1 recibió 0,1 ml de solución salina normal a través de los gavajes gástricos, el grupo 2 de ratas diabéticas control recibió una inyección intraperitoneal (IP) de STZ 60 mg / kg de peso corporal. Las ratas con FBS> 250 mg / dl se consideraron como diabéticas. El Grupo 3 eran ratas diabéticas que recibieron insulina en dosis de 3 U/ 100 g de peso corporal y los Grupos 4 y 5 estaban compuestos por ratas diabéticas que recibieron 0,1 cc con 200 y 400 mg / kg, de extracto de etanol de semillas de Lepidium sativum por gavajes diarios. Un día después de los últimos gavages, las ratas fueron anestesiadas con cloroformo. Se extrajo el epidídimo y se pesó con una pesa digital. Posteriormente, las muestras se pusieron en solución de Bouin para el estudio histológico. La administración de dosis de 200 y 400 mg / ml de extracto de semilla Lepidium sativum aumentó la altura del epitelio y disminuyó significativamente la densidad volumétrica intersticial y el grosor fibromuscular. Además, la densidad volumétrica del epitelio fibromuscular, lumen e intersticio disminuyeron significativamente. El diámetro tubular y el lumen no cambiaron significativamente en los diferentes grupos. El extracto de semilla de Lepidium sativum es un agente complementario beneficioso protector contra los efectos adversos de la diabetes en el sistema reproductor masculino.

Efecto protector del extracto alcaloidal de Lepidium meyenii ecotipo amarillo sobre la osteoporosis en ratas ovariectomizadas / Protective effect of alkaloid extract of Lepidium meyenii yellow ecotype on osteoporosis in ovariectomized rats

Introducción: La osteoporosis es un trastorno del metabolismo óseo que predispone a fracturas con una alta carga de morbilidad asociada. Otros autores han comprobado experimentalmente la propiedad antiresortiva del extracto etanólico de los ecotipos rojo y negro de Lepidium meyenii, "maca", el cual incluye diversos metabolitos. Por ello, hace falta estudios que determinen el efecto de la fracción alcaloidal en la osteoporosis. Objetivo: Demostrar el efecto protector del extracto alcaloidal de Lepidium meyenii ecotipo amarillo sobre la osteoporosis inducida en ratas ovariectomizadas. Diseño: analítico, experimental incompleto. Participantes: 50 ratas. Lugar: Instituto de Patología de UNMSM, 2015. Método: división aleatoria en 5 grupos de 10 cada uno: operación simulada (sham) y ovarectomizadas con tratamiento: estradiol (40µg/Kg), extracto alcaloidal (EA) a dosis de 75mg/k y 100mg/k (maca I y II respectivamente), y sin tratamiento protector. Dosis diarias vía orogástrica por 8 semanas. Posteriormente se determinó densidad mineral ósea (DMO), marcadores óseos: fosfatasa alcalina (FA), osteocalcina, telopéptido amino terminal del colágeno (NTX), estradiol, calcio y fósforo en suero; y histomorfometría. Resultados: No se evidenciaron cambios significativos en la densitometría vertebral, a nivel del fémur se evidenció disminución en el grupo maca II. La FA (141,90 ± 33,58UI/L) y la osteocalcina (38,578 ± 10,403ng/ml) mostraron niveles superiores no significativos, el nivel de estradiol con el grupo maca II fue superior no significativo. Los niveles de calcio y fósforo no mostraron variación. En la histomorfometría el grupo maca II (58,030 ± 4683) mostró mayor grosor trabecular significativamente. Conclusiones: se evidenció efecto protector antiresortivo parcial en los cambios inducidos por la ovariectomización, sin embargo, no se indujo osteoporosis al no encontrarse variación en los niveles de DMO.

Studies on the chemical constituents of the anti-osteoporotic part of Lepidium meyenii Walp. produced in Heqing / 药学学报

This study was designed to investigate the chemical constituents of the anti-osteoporotic part of Lepidium meyenii Walp. (maca) produced in Heqing, Yunnan. Seven compounds were isolated from the n-BuOH extract of maca using combination of column chromatographies on MCI resin, silica gel, C18 bonded silica gel, and Sephadex LH-20, followed by semi-preparative HPLC and recrystallization. The purified compounds were identified on the basis of their physicochemical properties and spectral data as macaolidine (1), tryptophan (2), daucosterol (3), (3S)-1,2,3,4-tetrahydro-β-carboline-3-carboxylicacid (4), chlorogenic acid (5), luteolin (6), and hyperoside (7). Compound 1 is a new phenylacetamide alkaloid, and compounds 4-7 were isolated from Lepidium meyenii for the first time.

Cloning and expression analysis of C4H gene from Lepidium apetalum / 药学学报

Lepidium apetalum was used as an experimental material in this study. By analyzing the tran-scriptome data of L. apetalum and application of the specific primers, cDNA of cinnamate-4-hydroxylase (C4H) gene was isolated from L. apetalum and named as LaC4H (GenBank accession No. KX064050). Meanwhile, the bioinformatic analysis, prokaryotic expression, purification, tissue-specific expression analysis and expres-sion analysis after methyl jasmonate (MeJA) treatment were carried out. The results indicated that: ① The open reading frame (ORF) of LaC4H was 1 518 bp, which encoded a protein of 505 amino acid residues, with a predicted molecular mass of 57.73 kD. ② Bioinformatic analysis showed that LaC4H protein contained the conserved sequences of cytochrome P450 (CYP450) and 5 substrate recognition sites (SRSs) of CYP73A1, therefore LaC4H protein was a member of CYP450 superfamily. The phylogenetic analysis indicated that LaC4H protein showed the highest homology with C4H protein from cruciferous plants (such as AtC4H from Arabidopsis thaliana). ③ Through the construction of the prokaryotic expression vector pET-32a-LaC4H, the recombinant LaC4H protein was successfully expressed in E. coli BL21 (DE3) cells and the recombinant LaC4H protein was purified by Ni2+ affinity chromatography. ④ Real-time PCR analysis indicated that LaC4H was expressed in a high transcript level in stems, lower levels in leaves and flowers, the lowest level in roots. After MeJA treatment, the expression level of LaC4H in leaves was increased significantly to reach the highest level at 48 h. Furthermore, the expression levels of LaC4H were positively correlated with the flavonoids contents in leaves. The results of this study provide the fundamental information on LaC4H gene in the flavonoids biosyn-thesis pathway of L. apetalum.