DIVERSIDAD GENÉTICA Y MEJORAMIENTO DE PLANTAS MEDICINALES= Medicinal plants and improvement of medicinal herbs

Different Arctium lappa L. explants and the compounding proportions of different hormones were comparatively studied in tissue culture and thereafter a high-efficiency plant-regeneration system by in vitro A. lappa culture was established. On MS media contg. 2.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5-2.0 mg/L benzyladenine (BA), cotyledon and hypocotyl pieces could be induced to produce calli at 87%-100%. On MS media contg. 1.0-3.0 mg/L 1-naphthaleneacetic acid (NAA) and 0.5-2.0 mg/L BA, the calli could indirectly differentiate or the explants could directly differentiate to form adventitious buds, and the differentiation rate of the calli could reached as high as 100%. The hypocotyls differentiated at a higher rate than the cotyledons, and their direct differentiation rate amounted to 77.3% on MS medium contg. 1.0 mg/L NAA and 1.0 mg/L BA. A. lappa was histochem. found capable of regenerating by two approaches, organogenesis and embryogenesis. Well-growing adventitious buds were inoculated on 1/2 MS medium contg. 1.0 mg/L NAA and 1.0 mg/L indolebutyric acid (IBA) for their rooting and thereafter transplanted, and their survival rate reached 93.3%. It approx. took 13 wk to complete the transition from callus induction to in vitro plantlet transplantation to perlite matrix. The in vitro plantlets flowered in the following year and presented a normal morphol. during their growth.

ÚLTIMOS AVANCES EN LA QUÍMICA Y ACTIVIDADES BACTERIOLÓGICAS EN LAS PLANTAS MEDICINALES= Medicinal plants, last advances on chemistry and bacteria activities on the medicinal herbs

1) The authors have developed a novel hybrid two-dimensional counter-current chromatog. and liq. chromatog. (2D CCC ´ LC) system for the continuous purifn. of arctiin from crude ext. of Arctium lappa.  The first dimensional CCC column has been designed to fractionalize crude complex ext. into pure arctiin effluent using a one-component org./salt-contg. system, and the second dimensional LC column has been packed with macroporous resin for online adsorption, desalination and desorption of arctiin which was effluent purified from the first CCC dimension.  Thus, the crude arctiin mixt. has been purified efficiently and conveniently by online CCC ´ LC in spite of the use of a salt-contg. solvent system in CCC sepn.  As a result, high purity (more than 97%) of arctiin has been isolated by repeated injections both using the Et acetate-8% sodium chloride aq. soln. and butanol-1% sodium chloride aq. soln.  By contrast with the traditional CCC processes using multi-component org./aq. solvent systems, the present online CCC ´ LC process only used a one-component org. solvent and thus the solvent is easier to recover and regenerate.  All of used solvents such as Et acetate, n-butanol and NaCl aq. soln. are low toxicity and environment-friendly.  Moreover, the lower phase of salt-contg. aq. soln. used as mobile phase, only contained minor org. solvent, which will save much org. solvent in continuous sepn.  In summary, the authors' results indicated that the online hybrid 2D CCC ´ LC system using one-component org./salt-contg. aq. soln. is very promising and powerful tool for high-throughput purifn. of arctiin from fruits of A. lappa.

2) The traditional visual identification of Chinese herbs is not objective.  Mol. biol. techniques can be used to accurately identify the origin of each herbal species.  This can enable the further purifn. and control of important herbal species.  Mol. techniques involving polymerase chain reaction and sequencing were used to provide a relatively simple and objective means of identifying burdock at the species level.  This study proved that the length of the ITS1-5.8S rRNA-ITS2 sequence was 358 base-pairs (bp) for six types of Arctium lappa Linn (the following breeds: Pingtung Gueilai, General Snow, Japanese, Fengshan, Wholesaler, and Tainan).  Automatic sequencing anal. found that ITS DNA sequences for Pingtung Gueilai and Japanese breeds were the same, and the General Snow breed differed from them at its 277 bp.  This study used DNA sequencing to analyze the high specificity regions of A. lappa Linn ITS, originated in different parts of Taiwan, and discovered the breed identification single-nucleotide polymorphism at the 277 bp position for local differentiation.

3) Ethnopharmacol. relevance: Arctigenin, a bioactive constituent from dried seeds of Arctium lappa L. (Compositae) which has been widely used as a Traditional Chinese Medicine for dispelling wind and heat included in Chinese Pharmacophere, was found to exhibit anti-inflammatory activities but its mol. mechanism remains unknown yet.  To investigate the anti-inflammatory mechanism of arctigenin.  Cultured macrophage RAW 264.7 cells and THP-1 cells were used for the expts.  Griess assay was used to evaluate the inhibitory effect of arctigenin on the overprodn. of nitric oxide (NO).  ELISA was used to det. the level of pro-inflammatory cytokines including tumor necrosis factor alpha (TNF-alpha) and interleukin-6 (IL-6).  The inhibitory effect on the enzymic activity of cyclooxygenase-2 (COX-2) was tested by colorimetric method.  Western blot was used to detect the expression of inducible nitric oxide synthase (iNOS) and COX-2.  Arctigenin suppressed lipopolysaccharide (LPS)-stimulated NO prodn. and pro-inflammatory cytokines secretion, including TNF-alpha and IL-6 in a dose-dependent manner.  Arctigenin also strongly inhibited the expression of iNOS and iNOS enzymic activity, whereas the expression of COX-2 and COX-2 enzymic activity were not affected by arctigenin.  These results indicated that potent inhibition on NO, TNF- alpha and IL-6, but not COX-2 expression and COX-2 activity, might constitute the anti-inflammatory mechanism of arctigenin.  Arctigenin suppressed the overprodn. of NO through down-regulation of iNOS expression and iNOS enzymic activity in LPS-stimulated macrophage.

PARTE UTILIZADA= Used part: Raíz fresca, hojas. 

ACCIÓN FARMACOLÓGICA= Pharmacological action: Contra los forúnculos y granos en general, diurético, refrescantes y antireumático. 

COMPOSICIÓN QUÍMICA= Chemical composition: Contiene abundante inulina (30-50%), mucílagos, Ácidos fenólicos (ácidos cafeico, clorogénico, isoclorogénico y derivados del ácido cafeico: arctiína). Trazas de aceite esencial (fenilacetaldehído, benzaldehído, metoxi y metilpiracinas. Poliacetilenos (ácido arético, arctinona, arctinol, arctinal. Lactonas sesquiterpénicas tipo guayanólido. Fitosteroles: beta-sitosterol, estigmasterol. Compuestos insaturados: polienos, poliínos. Taninos. Sales potásicas. 

ZONA GEOGRÁFICA= Geografical zone: Uruguay. 

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Toda la planta

Usos tradicionales:

a) uso interno: trastornos reumáticos, digestión difícil, retención de líquido. La infusión se prepara con 1 cucharada de hojas para 1 litro de agua recién hervida: beber 1 taza 2 a 3 veces al día. La decocción (cocimiento) se prepara con 1 cucharada de raíz y tallo picado o triturado en 1 litro de agua fría que se deja reposar algunas horas y luego se hierve no más de 5 minutos y al final se cuela: beber 1 taza 3 veces en el día.

b) uso externo: enfermedades de la piel (heridas, úlceras y psoriasis), hemorroides. La misma infusión para uso externo en forma de compresas o lavados.

Efectos: bactericida, depurativo, diurético, anti-inflamatorio.

Origin:
Native to northern Europe; now found in western Himalayas, Kashmir and Simla.

Action:
Hypoglycaemic (plant extract caused reduction of blood sugar with an increase in carbohydrate tolerance).
Roots—inhibitory of tumour growth, cardiac stimulant, diuretic, spasmolytic.
Leaves and seeds—anticutaneous (used in psoriasis, seborrhoic eczema).

1) GONZALEZ, Matías ; LOMBARDO, Atilio ; VALLARINO, Aida. Plantas de la medicina vulgar del Uruguay. Montevideo : Talleres Gráficos, 1937. p. 25.

2) Fitoterapia : vademecum de prescripcion : plantas medicinales. 3ª. ed. Barcelona : Masson, 1999, p. 136-137.

3) HE, Wenting; HOU, Suiwen; WANG, Chongying. Tissue culture and high-efficiency plant regeneration of Arctium lappa. Xibei Zhiwu Xuebao. 2006, vol.26, nº2, p.319-324.
 
4) GUO, Mengzhe; LIANG, Junling; WU, Shihua. On-line coupling of counter-current chromatography and macroporous resin chromatography for continuous isolation of arctiin from the fruit of Arctium lappa L. Journal of Chromatography, A. 2010, vol.1217, nº33, p.5398-5406.
 
5) CHANG, Hui-Jen, et al. Identification and authentication of burdock (Arctium lappa Linn) using PCR sequencing. Fooyin Journal of Health Sciences. 2009, vol.1, nº1, p.28-32.
 
6) ZHAO, Feng; WANG, Lu; LIU, Ke. In vitro anti-inflammatory effects of arctigenin, a lignan from Arctium lappa L., through inhibition on iNOS pathway. Journal of Ethnopharmacology. 2009, vol.122, nº3, p.457-462.

7) ALONSO, Jorge R. Tratado de fitomedicina : bases clínicas y farmacológicas. Buenos Aires : ISIS, 1998, p. 292.

8) Hierbas medicinales/ Chile. Ministerio de Agricultura.  p.64

9) Khare, C.P./ Indian Medicinal Plants. -- Nueva Dheli: Springer, 2007 . - p. 836.