Isolation and purification of 5-hydroxytryptophan in extract from seeds of Griffonia simplicifolia by ion-exchange resin / 中草药

Objective: To research the adsortpion and separation function of ion-exchange resin on 5-hydroxytryptophan (5-HTP) in extract from the seeds of Griffonia simplicifolia and to screen the ion-exchange resin with high selective adsorption. Methods: The static adsorptive properties of eight different types of ion-exchange resins were evaluated using adsorptive content, desorption rate, adsorption rate, and adsorption temperature as indexes in order to optimize the resin. And the effects of different pH values, injection rates, injection concentration, eluent concentration, and elution rates on adsorptive properties of resins were investigated. Results: The 001 × 7 cation exchange resin showed the best comprehensive adsorption property. The loading solution concentration was 10.8 mg/mL, the pH value was 3.5, the flow rate of loading solution was 4.0 mL/min, and 3BV 7.0% ammonia water solution in 3.0 mL/min velocity was used to elute. In this process, the product was obtained with the 5-HTP purity of greater than 99.0% and the ash content of less than 1.0%, product average yield was 7.95% and the product quality could meet the market demand. Conclusion: The 001 × 7 cation exchange resin shows a better comprehensive adsorption property. It could be used to isolate and purify the 5-HTP for scale production.

A new β-carboline alkaloid from the seeds of Griffonia simplicifolia / 中国天然药物

AIM@#To study the chemical constituents from the seeds of Griffonia simplicifolia Baill.@*METHODS@#The chemical constituents were isolated and purified by a combination of chromatographic materials including silica gel, Sephadex LH-20, and preparative TLC. Their structures were elucidated on the basis of MS and NMR data analysis. The cytotoxic activities were evaluated against HepG2 cancer cell line using the MTT colorimetric method.@*RESULTS@#A new β-carboline alkaloid, griffonine (1), together with seven known alkaloids, hyrtioerectine B (2), 3-carboxy-6-hydroxy-β-carboline (3), hyrtiosulawesine (4), 5-hydroxyindole-3-carbaldehyde (5), 5-hydroxy-3-(2-hydroxyethyl) indole (6), trigonelline (7), and 5-hydroxytryptamine (8) were isolated and identified. Alkaloids 1, 2 and 4 showed growth inhibitory effects on the HepG2 cell line with IC50 values of 23.5, 9.6 and 19.3 μmol·L(-1), respectively.@*CONCLUSION@#Alkaloid 1 is new and was named griffonine. Alkaloids 2-7 were isolated from this plant for the first time. Alkaloids 1, 2 and 4 were potentially cytotoxic.

Degeneration of primary afferent terminals following brachial plexus extensive avulsion injury in rats / Degeneración de los terminales aferentes primarios de rata luego de lesión extensa por avulsión del plexo braquial

Important breakthroughs in the understanding regeneration failure in an injured CNS have been made by studies of primary afferent neurons. Dorsal rhizotomy has provided an experimental model of brachial plexus (BP) avulsion. This is an injury in which the central branches of primary afferents are disrupted at their point of entry into the spinal cord, bringing motor and sensory dysfunction to the upper limbs. In the present work, the central axonal organization of primary afferents was examined in control (without lesion) adult Wistar rats and in rats subjected to a C3-T3 rhizotomy. Specific sensory axon subtypes were recognized by application of antibodies to the calcitonin gene-related peptide (CGRP), the P2X3 purinoreceptor, the low-affinity p75-neurotrophin receptor and the retrograde tracer cholera toxin subunit beta (TCbeta ). Other subtypes weres labeled with the lectin Griffonia simplicifolia IB4. Using immunohistochemistry and high resolution light microscopy, brachial plexus rhizotomy in adult rats has proven a reliable model for several neural deficits in humans. This lesion produced different degrees of terminal degeneration in the several types of primary afferents which define sub-populations of sensitive neurons. Between the C6 and C8 levels of the spinal cord,,deafferentation was partial for peptidergic GCRP-positive fibers, in contrast with elimination of non peptidergic and myelinated fibers. Dorsal rhizotomy has provided an adequate experimental model to study sensory alterations such as acute pain and allodynia as well as factors that affect regeneration into the CNS., Therefore, the differential deafferentation response must be considered inr the evaluation of therapies for nociception (pain) and regeneration for brachial plexus avulsion. The anatomical diffierences among the primary afferent subtypes also affect their roles in normal and damaged conditions.

El uso de las neuronas sensoriales primarias ha aportado avances en el entendimiento de las razones por las cuales falla la regeneración cuando el sistema nervioso central (SNC) es dañado. La rizotomía dorsal se puede usar como un modelo experimental de las lesiones por avulsión del plexo braquial, una lesión en la cual son desprendidas, en su punto de entrada en la médula espinal, las ramas centrales de los aferentes primarios causando una disfunción motora y sensorial grave e irreversible del miembro superior. En el presente trabajo, se examinó la organización central de los aferentes primarios en ratas Wistar adultas. Éstas fueron divididas en controles normales no lesionados y en animales rizotomizados entre los niveles cervical 3 y torácico 3 (C3-T3). Se estudió la deaferentación de los subtipos de axones sensoriales utilizando anticuerpos específicos contra el péptido relacionado con el gen de la calcitonina (CGRP), el receptor purinérgico (P2X3), el receptor de baja afinidad p75 para el factor de crecimiento nervioso (NGF) y contra la subunidad ®de la toxina de cólera (TCbeta ). Otro subtipo fue marcado con la lectina Griffonia simplicifolia IB4. La inmunohistoquímica y la microscopía óptica de alta resolución demostraron que el modelo animal de rizotomía completa del plexo braquial reproduce diversos déficit observados en las lesiones humanas. Esta lesión produce diferentes grados de degeneración terminal entre los diversos tipos de aferentes primarios que definen subpoblaciones de neuronas sensoriales. En los niveles de la médula espinal estudiados (entre C6 y C8), la deaferentación fue parcial para las fibras peptidérgicas GCRPpositivas, en contraste con la eliminación de las fibras no peptidérgicas y las mielinizadas. La rizotomía dorsal es un modelo experimental apropiado para estudiar las alteraciones sensoriales como el dolor agudo y la alodinia, así como los factores que podrían afectar la regeneración en el SNC. Por tanto, la respuesta de deaferentacion diferencial debe ser tenida en cuenta para la evaluación de terapias antinociceptivas y regenerativas tras la avulsión del plexo braquial. Se discute la anatomía de los subtipos de aferentes primarios y su papel en condiciones normales y después de la lesión.

Interaction and in vivo growth inhibition of Ehrlich ascites tumor cells by jacalin.

Jacalin has been found to agglutinate Ehrlich ascites cells. The agglutination was inhibited by a-glycosides of D-Gal and ß -D-Gal(1 → 3)-D-GalNAc suggesting that the lectin-ascites interaction was carbohydrate-specific. There was 21·8% inhibition of tumour (ascites) cell growth in vivo in mice administered 50μg of jacalin by injection for 6 days following intraperitoneal injection of ascites cells. Administration of 100, 150 and 200 μg jacalin resulted in 40·2, 57·5 and 83% inhibition respectively. The in vivo inhibition of tumour cells growth by jacalin was due to its preferential binding with D-Gal-a -( 1→6) present as terminal residues in the glycoprotein on tumour cell surface.

Lectins from Griffonia simplicifolia seeds.

The physical-chemical and carbohydrate binding specificity of Griffonia simplicifolia I (GS I) isolectins, one of the 4 lectins isolated from Griffonia simplicifolia seeds, are described. Association constants for the binding of methyl α- and β-D-galactopyranoside and methyl 2-acetamido-2-deoxy-α-D-galactopyranoside to the A4, A2 B2 and B4 isolectins are reported. Precipitation reactions of the Griffonia simplicifolia isolectins with guaran and type Β blood group substance are described. The hypothesis that subunit Β is a precursor of subunit A, a process involving proteolytic cleavage of the Β subunit, was tested by conducting structural studies on the 2 subunits. The results indicated that the A and Β subunits are probably products of 2 separate but closely related, possibly contiguous genes.