ABSTRACT
This study aimed to determine the mimosine level and examine the male reproductive toxicity effects of Leucaena leucocephala (LL) shoot tips plus young leaf extract. Mimosine level in LL extract was determined by thin layer chromatography before administration in animals. Male rats were divided into control and LL (1,500 mg/KgBW) groups (n = 6). After 60 days of experiment, serum sex hormones, sperm quality, and testicular histopathology were assayed and observed. Malondialdehyde (MDA) level and expressions of steroidogenic acute regulatory (StAR) and phosphorylated proteins in testicular lysate were examined by western blotting. Results showed that mimosine levels in LL extract was 17.35 ± 1.12 % of dry weight. LL significantly decreased FSH & LH levels, sperm qualities, and seminiferous tubule diameter compared to the control (p<0.05). Seminiferous tubular atrophies, germ cell sloughing, and degenerations were observed in LL group. In addition, testicular MDA level and StAR protein expression were significantly decreased in LL group. LL extract could increase the expression of a 50 kDa phohorylated protein in testicular lysate. In conclusion, LL extract has mimosine and reproductive toxicity effects on males.
Este trabajo tuvo como objetivo determinar el nivel de mimosina y examinar los efectos de la toxicidad reproductiva de los brotes de Leucaena leucocephala (LL), más el extracto de hojas jóvenes, en ratas macho. El nivel de mimosina en el extracto de LL se determinó mediante cromatografía en capa fina antes de la administración en animales. Las ratas se dividieron en grupos de control y LL (1,500 mg / kgBW) (n = 6). Después de 60 días, se analizaron y observaron las hormonas sexuales séricas, la calidad de los espermatozoides y la histopatología testicular. A través de Western Blot se examinaron el nivel de malondialdehído (MDA), las expresiones de reguladores agudos esteroidogénicos (StAR) y las proteínas fosforiladas en el lisado testicular. Los resultados mostraron que los niveles de mimosina en el extracto de LL fueron 17.35 ± 1.12 % del peso seco. LL disminuyó significativamente los niveles de FSH y LH, la calidad de los espermatozoides y el diámetro de los túbulos seminíferos en comparación con el control (p <0,05). Se observaron atrofias en los túbulos seminíferos, desprendimiento de células germinales y degeneraciones en el grupo LL. Además, el nivel de MDA testicular y la expresión de la proteína StAR se redujeron significativamente en el grupo LL. El extracto de LL podría aumentar la expresión de la proteína fosforilada de 50 kDa en el lisado testicular. En conclusión, el extracto de LL tiene mimosina y efectos de toxicidad reproductiva en los hombres.
Subject(s)
Animals , Male , Rats , Plant Extracts/toxicity , Plant Extracts/chemistry , Genitalia, Male/drug effects , Fabaceae , Mimosine/analysis , Sperm Count , Spermatozoa/drug effects , Testis/drug effects , Blotting, WesternABSTRACT
This study aimed to determine the antioxidant capacity of the Leucaena leucocephala aqueous shoot tips plus young leaves (LL-spl) extracts among three different fractions (LL-spl 10, 20, and 40 min) and to examine its acute toxicity on male reproductive parameters. The amount of the total phenolics in LL-spl extract was determined using a Folin-Ciocalteu reagent method and its antioxidant capacity was analyzed using 1, 1-diphenyl l-2-picrylhydrazyl radical scavenging and ferric reducing antioxidant powder methods. The LL-spl extract fraction with highest antioxidant capacity was used in animal treating. Male rats were divided into three groups (n= 5); control and groups treated with LL-spl 400 and 600 mg/Kg body weight for consecutive 40 days. The results showed that the LL-spl 40 min fraction possessed the highest antioxidant capacity. In addition, the LL-spl 400 and 600 groups showed no differences in weights of body, testis and epididymis, serum testosterone levels, and expression of testicular steroidogenic acute regulatory (StAR) protein. Significantly, LL-spl extract reduced the weight of seminal vesicle, sperm concentration, and seminiferous diameters compared with control. Moreover, LL-spl extract had adverse effect on testicular histology in inducing of seminiferous atrophy and degeneration including dilated blood vessels in interstitial tissue. It was concluded that although LL-spl extract possessing antioxidant capacity, in short term consumptions, it could be toxic to some male reproductive organs especially damaging testicular tissues.
El objetivo fue determinar la capacidad antioxidante del extracto de brotes acuosos con hojas nuevas de Leucaena leucocephala (LL-spl) en tres fracciones diferentes (LL-SPL 10, 20 y 40 min), además de examinar su toxicidad aguda sobre los parámetros reproductivos masculinos. Se determinó la cantidad de los fenoles totales en el extracto de LL-spl utilizando un método reactivo de Folin-Ciocalteu. La capacidad antioxidante se analizó por medio de 1-difenil-2-picrilhidracilo y/o métodos de reducción férrica de la capacidad antioxidante. La fracción de extracto LL-spl con mayor capacidad antioxidante fue utilizada en el tratamiento de los animales. Ratas macho fueron divididas en tres grupos (n= 5): el control y los grupos tratados con LL-spl 400 y 600 mg/kg peso corporal por 40 días consecutivos. El resultado mostró que la fracción LL-spl 40 min poseía la mayor capacidad antioxidante. Además, los grupos 400 y 600 LL-spl no mostraron diferencias según el peso corporal, testículos y epidídimo, niveles de testosterona y la expresión de proteínas testiculares. El extracto de LL-spl redujo de manera significativa el peso de la vesícula seminal, la concentración de espermatozoides y los diámetros de los túbulos seminíferos en comparación con el control. Por otra parte, el extracto de LL-spl tuvo un efecto adverso sobre la histología testicular por la inducción de atrofia y degeneración de los túbulos seminíferos, incluyendo a vasos sanguíneos dilatados en el tejido intersticial. Si bien el extracto LL-spl posee una capacidad antioxidante, ésta podría ser tóxica en el consumo a corto plazo para algunos órganos reproductores masculinos y especialmente dañino para los tejidos testiculares.
Subject(s)
Animals , Male , Rats , Antioxidants/pharmacology , Fabaceae , Plant Extracts/pharmacology , Plant Leaves/chemistry , Testis/drug effects , Body Weight/drug effects , Genitalia, Male/drug effects , Phenols/analysis , Rats, Sprague-Dawley , Seminiferous Tubules , Testosterone/analysis , Toxicity Tests, AcuteABSTRACT
Protocolos eficientes de crescimento de ápices caulinares de alho (Allium sativum L.) e posterior bulbificação in vitro são importantes para limpeza clonal e manutenção da fidelidade genética. O trabalho teve como objetivo avaliar os efeitos de tipos e concentrações de reguladores de crescimento sobre a morfogênese de plantas de alho in vitro. Ápices caulinares com até dois primórdios foram excisados de bulbilhos de alho da cv. 'Jonas' e submetidos ao cultivo in vitro em meio de cultura suplementado de ácido indolacético (0; 1,07; 2,69 e 5,37µM), ácido indolbutírico (0; 0,49; 0,98 e 2,46µM), ácido naftalenoacético (0; 1,07; 2,69 e 5,37µM), ácido jasmônico (0; 1,0; 5,0 e 10,0µM) e ácido abscísico (0; 0,38; 1,89; e 3,78µM). A concentração de 1,07µM de ácido naftalenoacético aplicado ao meio de cultura promoveu incrementos na maioria das variáveis analisadas. O ácido jasmônico induziu a formação de bulbos de alho in vitro, embora tenha apresentado performance inferior ao verificado com o uso de ANA. Por outro lado, a adição de ácido abscísico no meio de cultura inibiu o crescimento de plantas, porém, não impediu a formação de bulbos, sobretudo em concentrações reduzidas. De um modo geral, as variáveis número de bulbos e porcentagem de bulbificação diminuiram com o uso de concentrações elevadas dos reguladores de crescimento testados. Entre os reguladores de crescimento de plantas, o ANA apresenta maior efeito na morfogênese in vitro de plantas de alho, entretanto, o ácido jasmônico e o ABA também apresentam potencial para induzir a formação de bulbos de alho in vitro como o ANA.
Efficient protocols for garlic (Allium sativum L.) shoot tips growth and later in vitro bulbing are significant for pathogen removal and maintenance of genetic fidelity. The objective of this study was to assess the effects of different types and concentrations of growth regulators on in vitro morphogenesis of garlic plants. Shoot meristems with up to two primordial were excised from garlic bulbils (cv. 'Jonas') and cultivated in vitro in culture media supplemented with indoleacetic acid (0; 1,07; 2,69 e 5,37µM), indolbutyric acid (0; 0,49; 0,98 e 2,46µM), naphthaleneacetic acid (0; 1,07; 2,69 e 5,37µM), and abscisic acid (0; 0,38; 1,89; e 3,78µM). The concentration of 1,07µM indoleacetic acid applied to the culture medium promoted increases in most variables analyzed. Jasmonic acid induced formation of garlic bulbs in vitro, although it showed lower performance than that verified with the use of NAA. On the other hand, addition of abscisic acid in culture media inhibited plant growth. However, it did not impede the formation of bulbs, especially when in reduced concentrations. Generally speaking, variables such as number of bulbs and rate of bulbing decreased with the use of high concentrations of the assessed growth regulators. Among plant growth regulators, NAA showed a stronger effect on in vitro morphogenesis of garlic plants. Nonetheless, jasmonic acid and ABA also showed potential to induce formation of garlic bulbs in vitro such as NAA.
ABSTRACT
Objective To get a new approach for conserving the germplasm of Siraitia grosvenorii Methods Shoots, 0.8—1 cm in length, excised from test-tube plantlets which were subcultured 15 d, were precultured. Then its shoot-tips about 2—3 mm in length were dissected and loaded with 60% PVS2 at 25 ℃, and dehydrated with 100% PVS2 at 0 ℃, changed for fresh 100% PVS2 prior to directly plunging into liquid nitrogen. After cryopreservation for 24 h, the shoot tips were thawed, rinsed in MS+1.2 mol/L sucrose medium, blotted with filter papers, then plated on the MS + 1.0 mg/L 6-BA+0.05 mg/L NAA + 0.1 mg/LGA3 + 0.8% agar + 45 g/L sucrose medium at 25 ℃ for 7 d in dark prior to exposure to the light. The root medium for regeneration plantlets was 1/2 MS + 0.2 mg/L NAA + 30 g/L sucrose. Results Shoots were precultured for 3 d on MS + 0.7 mol/L sucrose medium, then its shoot-tips loaded for 40 min, dehydrated for 50 min. After cryopreservation, the shoot tips were rapidly thawed in water at 40 ℃, then rinsed for 40 min. The survival rate of shoot-tips plated on the recovering medium in one week was 100%, and regeneration rate after 30 d was 78.33%, which was the highest. The regeneration plantlets inoculated on root medium were reconstructed integrating plantlets. Conclusion The method of vitrification to cryopreserve the germplasm of S. grosvenorii is a simple way with high survival rate and normal regeneration plants.