Hepatoprotective activity of Zha Xun and its different solvent-eluting components / 药学学报

A pharmacophore-based study was conducted to investigate the therapeutic activity of the traditional Tibetan medicine Zha Xun (ZX) in liver diseases. In the present study, the protective effect of ZX on the acute liver injury induced by concanavalin A (ConA) and 0.15% carbon tetrachloride (0.15% CCl4) in ICR mice was evaluated, and the results showed that ZX significantly reduced serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the ConA-induced acute immune liver injury model and the CCl4-induced acute oxidative liver injury model (P < 0.05). Subsequently, the protective effects of aqueous, 95% ethanol, 60% ethanol and 30% ethanol eluting fractions of ZX, and fulvic acid, the main water-soluble constituent of ZX, were evaluated against acute oxidative liver injury induced by 0.15% CCl4 in mice. The results showed that different solvent-eluting fractions of ZX showed certain hepatoprotective activities, among which the aqueous extract of ZX and 30% ethanol extract of ZX significantly reduced the serum levels of ALT, AST, and lactate dehydrogenase (LDH) in mice (P < 0.05), and the serum levels of LDH in mice were significantly reduced by fulvic acid (P < 0.05), which showed significant hepatoprotective activity. The protective activities and preliminary mechanisms of the total extract of ZX, the aqueous extract of ZX, the 30% ethanol extract of ZX, and fulvic acid against hepatocellular injury in vitro were further evaluated by using the H2O2-induced hepatocellular injury model. The results showed that the components could significantly inhibit H2O2-induced hepatocellular injury, reduce the levels of ALT, alkaline phosphatase (ALP), and LDH, improve the survival rate of hepatocellular cells, and reduce the content of intracellular reactive oxygen species (ROS) in cell culture. At the same time, it can inhibit hepatocyte apoptosis by increasing the expression ratio of Bcl-2/BAX protein and decreasing the expression ratio of cleaved caspase-3/pro caspase-3 protein. The present study showed that ZX has clear hepatoprotective activity in vitro and in vivo, and the different solvent elution fractions of ZX showed certain hepatoprotective activity, among which the aqueous extract of ZX, 30% ethanol extract of ZX had better hepatoprotective activity, and the activity of 60% ethanol extract of ZX was stronger than that of 95% ethanol extract of ZX. The activity of ZX and its water-soluble elution site exerted hepatoprotective effects by inhibiting hepatocyte apoptosis and oxidative stress. The animals used in this experiment and related disposal meet the requirements of animal welfare, and have been reviewed and approved by the Laboratory Animal Management and Use Committee of the Institute of Materia Medica, Chinese Academy of Medical Sciences (approval number: 00004018).

Bioavailability augmentation of silymarin using natural bioenhancers: An in vivo pharmacokinetic study

Abstract Pharmacokinetic studies were carried out in male and female rats to quantify silymarin as silybin (A+B) after the oral administration of various silymarin formulations combined with three bioenhancers, namely, lysergol, piperine, and fulvic acid, and compared with plain silymarin formulation (control). A non-compartmental analysis, model independent analysis, was utilized, and various pharmacokinetic parameters (C max, T max, and AUC 0-t) were calculated individually for each treatment group, and the values were expressed as mean ± SEM (n = 6). Plasma samples obtained from the rats were analyzed for the concentration of silymarin through a validated RP-HPLC method and on the basis of data generated from the pharmacokinetic studies. Results indicated that the bioenhancers augmented pharmacokinetic parameters and bioavailability increased 2.4-14.5-fold in all the formulations compared with the control. The current work envisages the development of an industrially viable product that can be further subjected to clinical trials and scientifically supports the development of silymarin as a contemporary therapeutic agent with enhanced bioavailability and medicinal values.

Recalcitrant carbon and nitrogen in agriculture soi ls with residue accumulation and fertilization under tropical conditions / Carbono recalcitrante e nitrogênio em solos com acumulação e fertilização de resíduos em condições tropicais

Soil organic matter has a strong relation to total organic carbon, and about 85% of organic carbon consists of humic substances (HS), classified as humin (HU), humic (HA) and fulvic acids (FA), and denominated as recalcitrant carbon in soil. HS are formed by complex, heterogeneous and polydisperse molecules, which have significant influence on the soil physical and chemical characteristics. The study evaluates the hypothesis that agricultural soils treated with organic residues may present higher carbon stocks as presented in forest soils. The aim of this study was to evaluate alterations in recalcitrant carbon and nitrogen stocks in Oxisol (Forest - FL, unfertilized Brachiaria - UB, and fertilized Brachiaria - FB) and Cambisol (Forest - FC, Coastcross - CC, sugarcane - CA, and silage corn - SM) at surface (0.0 - 0.1) and subsurface (0.1 - 0.2 m), in the Zona da Mata in Minas Gerais state, Brazil. Result shows that fertilization, low soil disturbance and residual removal promoted increase of C and N content in HS, being close to native forests. Both, carbon and nitrogen recalcitrant, presented reduction with soil depth. HU ranged from 10.5 to 16.7 g kg-1 and presented the highest concentration compared to FA and HA. In Oxisol, FL and FB presented the highest SH concentration and demonstrate the positive effect of fertilization on carbon stocks improving soil quality in well-managed and productive areas. While, In Cambisol, FC and CC presented higher carbon and nitrogen in HS, mainly for HU and HA fractions, and SM showed the lowest concentrations in all fractions. Our results suggest that soil managements with lower soil disturbance and residual removal promotes increasing of carbon and nitrogen in recalcitrant fraction, with concentration close to native forests. Pasture should be fertilized to improve recalcitrant carbon and nitrogen stocks, avoiding process of degradation in tropical soil. It is an important outcome due to high levels of degraded areas in Brazil caused by inadequate use of soil mainly with pasture.

A matéria orgânica do solo tem uma forte relação com o carbono orgânico total, cerca de 85% do carbono orgânico é composto por substâncias húmicas (HS), classificadas como humina (HU), ácidos húmicos (HA) e fúlvicos (FA), denominadas como carbono recalcitrante no solo. As HS são formadas por moléculas complexas, heterogêneas e polidispersas, que exercem influência significativa nas características físicas e químicas do solo. O estudo avalia a hipótese de que solos agrícolas tratados com resíduos orgânicos podem apresentar maiores estoques de carbono como os encontrados em solos florestais. O objetivo destetrabalho foi avaliar as alterações nos estoques de carbono e nitrogênio recalcitrantes em Latossolo (Floresta - FL, Brachiaria não fertilizada ­ UB, e Brachiaria fertilizada - FB) e Cambissolo (Floresta - FC, coastcross - CC, cana-de-açúcar e milho para silagem - SM) na superfície (0,0 - 0,1) e sub-superfície (0,1 - 0,2 m), na Zona da Mata em Minas Gerais, Brasil. Os resultados mostram que a adubação, a baixa perturbação do solo e a remoção dos resíduos promoveram aumento do teor de C e N na HS, estando próximos às florestas nativas. Ambos, carbono e nitrogênio recalcitrantes, apresentaram redução com a profundidade do solo. A HU variou de 10,5 a 16,7 g kg-1 e apresentou a maior concentração em relação à FA e HA. Em Latossolo, FL e FB apresentaram a maior concentração de SH e demonstraram o efeito positivo da adubação nos estoques de carbono, melhorando a qualidade do solo em áreas bem manejadas e produtivas. Enquanto, no Cambissolo, FC e CC apresentarammaiores teores de carbono e nitrogênio nas HS, principalmente nas frações HU e HA, e SM apresentou as menores concentrações em todas as frações. Nossos resultados mostram que manejos de solo com menor perturbação e remoção de resíduos promovem o aumento de carbono e nitrogênio em frações recalcitrantes, com concentração próxima às florestas nativas. A pastagem deve ser fertilizada para melhorar o estoque de carbono e nitrogênio recalcitrante, evitando o processo de degradação do solo tropical. Este é um resultado importante devido aos altos níveis de áreas degradadas no Brasil causadas pelo uso inadequado do solo,principalmente com pastagens.

Efeito da adubação orgânica sobre as frações de carbono de solos cultivados com alfacen americana / Organic fertilizer effects upon carbon fractions from soils cultivated with iceberg lettuce

Avaliou-se, neste estudo, o efeito da aplicação de adubos orgânicos, minerais e calagem, nas frações húmica, fúlvica e nos teores de carbono orgânico de um Latossolo Vermelho e de um Cambissolo, cultivados com alface (americana). O experimento, realizado no Departamento de Ciência do Solo da Universidade Federal de Lavras, foi constituído de cinco doses de material húmico (0, 20, 40, 100 e 200 L ha-1), três tipos de adubação (composto orgânico, esterco de aves e mineral) e calagem (com e sem uso de calcário), em cinco repetições. No Cambissolo, a calagem contribuiu para a diminuição do teor de C orgânico nas áreas adubadas com fertilizante mineral e composto, e aumentou o armazenamento de C no solo adubado com esterco de aves. Em relação à adubação mineral, o uso de adubo orgânico aumentou os teores de C-fração ácido húmico e a relação C-húmico/fúlvico. No Latossolo, independente do tipo de adubação, o uso da calagem propiciou o armazenamento no solo de mais C do que o observado na área sem correção da acidez. Os efeitos da calagem e das fontes de nutrientes variaram em função da fração orgânica analisada ou das relações entre seus teores de carbono.

This study evaluated the effects of organic and mineral fertilizers and liming, on humic, fulvic, and organic carbon fractions of an Inceptisol and an Oxisol grown with lettuce. The experiment was carried out at the Soil Science Department of Federal University of Lavras, Minas Gerais state, Brazil. It was performed with 5 levels of soil conditioner (0, 20, 40, 100, 200 L ha-1), 3 fertilizers (organic compost, chicken manure and mineral), and liming (present or absent) using five replications. In the Inceptisol essay, the liming contributed to a decrease in the organic-C content in the mineral and compost fertilized plots, and increased C-accumulation in the treatments under chicken manure. The organic fertilizer increased the content of C-humic acid and the relation Chumic/fulvic. In the Oxisol essay, the liming contributed to C-accumulation when related to the treatments without liming. Liming effects and fertilizers varied as a function of the analyzed organic fractions and C content.

Caracterizacióndde ácidos húmicos y fúlvicos de un mollisol bajo dos coberturas difreentes / Charaterization of humic and fulvic acids from a mollisol under two diferent coverlands

Para evaluar y comparar algunas propiedades de los ácidos húmicos (AH) y fúlvicos (AF) como el grado de condensación y polimerización, el contenido total de carbono, hidrógeno y nitrógeno, y sus constantes de acidez, se analizó la fracción menor a 50 µm de un Mollisol del horizonte Ap, bajo caña de azúcar, y se comparó con un horizonte A, bajo bosque. Para tal fin se hicieron extracciones sucesivas con tetraborato de sodio 0,05 M, pirofosfato de sodio 0,025 M e hidróxido de sodio 0,1M. Las sustancias húmicas se purificaron mediante tratamientos con ácidos inorgánicos en solución al 1% de HCl y HF, resinas de absorción Amberlita XAD-16 e intercambio catiónico R101H+, diálisis y liofilización, y se caracterizaron por análisis elemental, relación E4/E6 y potenciometría ácido-base de acuerdo con las funciones de Gran modificadas. La relación E4/E6 permitió establecer mayores tamaños moleculares y grado de condensación de los ácidos húmicos en relación con los ácidos fúlvicos. Por potenciometría ácido-base se determinaron, en los ácidos húmicos y ácidos fúlvicos, las concentraciones de grupos funcionales ácidos y sus respectivas constantes de acidez. Dos de éstas, tituladas entre pH 4,0 y 6,1 corresponden a grupos carboxílicos, las otras dos entre pH 7,2 y 8,7 son el resultado de grupos -OH fenólicos fuertes y carboxílicos débiles. En el horizonte Ap, bajo caña de azúcar, los resultados demostraron que las prácticas de uso y manejo de este cultivo han llevado a la homogeneización de las sustancias húmicas, mientras que en el horizonte A bajo bosque existen ácidos húmicos y fúlvicos diferenciados por tamaño molecular, aromaticidad y grado de humificación.

In order to determine and to compare some physicochemical properties of humic (HA) and fulvic (FA) acids such as degree of condensation and polymerization, the total C,H,N content and pKa values were analyzed in the smaller fraction (< 50 m) from Ap horizon under sugarcane crop in opposition to A horizon under forest, both Mollisol order. To carry out this aim, successive extractions with sodium tetraborate 0.05 M, sodium pyrophosphate 0.025 M, and sodium hydroxide 0.1 M were made. The purification of these humic substances by treatment with inorganic acids 1% solutions of HCl and HF, Amberlite XAD-16 adsorption resin and R 101H+ cationic exchange, dialysis and freeze driying were done. The humic and fulvic acids were characterized by elemental analysis, E4/E6 relation and acid-base potentiometry. The ratio E4/E6 allows to establish that molecular size and degree of condensation in humic acids were bigger than in fulvic acids. By means of acid-base potentiometry, the concentrations of functional acid groups and their acid constants in the humic and fulvic acids were determined. Four constants were found; first and second between pH 4.0 and 6.1 correspond to differents kinds of carboxyl groups and another two between pH 7.2 and 8.7 come from strong O-H phenolic and weak carboxyl groups. In the Ap horizon under sugarcane the characterization showed that the handling and land use practices promote the homogeneity of humic substances meanwhile in the A horizon under forest; the humic and fulvic acids are different due to their degree of condensation molecular size and humification index.