Estrogenic and mutagenic activities of Crotalaria pallida measured by recombinant yeast assay and Ames test.

BACKGROUND: Crotalaria pallida Ailton is a plant belonging to the Fabaceae family, popularly known as "rattle or rattlesnake" and used in traditional medicine to treat swelling of the joints and as a vermifuge. Previous pharmacological studies have also reported anti-inflammatory, antimicrobial and antifungal activities. Nevertheless, scientific information regarding this species is scarce, and there are no reports related to its possible estrogenic and mutagenic effects. Thus, the purpose of the present study was to investigate the estrogenic potential of C. pallida leaves by means of the Recombinant Yeast Assay (RYA), seeking an alternative for estrogen replacement therapy during menopause; and to reflect on the safe use of natural products to assess the mutagenic activity of the crude extract from C. pallida leaves, the dichloromethane fraction and stigmasterol by means of the Ames test. METHODS: The recombinant yeast assay with the strain BY4741 of Saccharomyces cerevisiae, was performed with the ethanolic extract, dichloromethane fraction and stigmasterol isolated from the leaves of C. pallida. Mutagenic activity was evaluated by the Salmonella/microsome assay (Ames test), using the Salmonella typhimurium tester strains TA100, TA98, TA97 and TA102, with (+S9) and without (-S9) metabolization, by the preincubation method. RESULTS: All samples showed estrogenic activity, mainly stigmasterol. The ethanolic extract from C. pallida leaves showed mutagenic activity in the TA98 strain (-S9), whereas dichloromethane fraction and stigmasterol were found devoid of activity. CONCLUSION: Considering the excellent estrogenic activity performed by stigmasterol in the RYA associated with the absence of mutagenic activity when evaluated by the Ames test, stigmasterol becomes a strong candidate to be used in hormone replacement therapy during menopause.

Aspectos clínico-patológicos da intoxicação experimental pelas sementes de Crotalaria mucronata (Fabaceae) em bovinos / Experimental poisoning by Crotalaria mucronata (Fabaceae) seeds in cattle

Sementes moídas de Crotalaria mucronata Desv. (=Crotalaria striata DC., Crotalaria pallida Ait., Crotalaria saltiana), com nomes populares de "xique-xique" ou "guizo-de-cascavel", foram administradas por via oral a 9 bovinos. As doses diárias de 1g/kg, 2g/kg, 3g/kg cada uma a um bovino, e 5g/kg em dois de três bovinos, dadas durante 61 a 63 dias, não causaram intoxicação. A dose de 5g/kg em um bovino, 7,5g/kg em dois bovinos e 10g/kg em um bovino, dadas durante 47-61 dias, causaram sintomas entre 47 e 80 dias após o início da administração e a morte entre 3 horas e 5 dias após o início dos sintomas. Os principais sintomas foram pulso venoso positivo da veia jugular, respiração abdominal, taquicardia, inapetência, fezes ressequidas, edema sub-mandibular e fraqueza. Os achados de necropsia foram palidez pulmonar, hidropericárdio, hidrotórax, hidroperitôneo, edema de mesentério, aumento da consistência hepática, alterações de cor do fígado, dilatação de ventrículo cardíaco direito e edema da parede ruminal. As principais alterações histológicas concentraram-se nos pulmões, sob forma de espessamento das paredes alveolares e da parede das arteríolas com diminuição da luz e fibrose periarteriolar; havia também lesões hepáticas e cardíacas de menor importância. Pode-se concluir que as principais lesões causadas pela ingestão das sementes de C. mucronata durante períodos prolongados, devem-se à dificuldade de passagem do sangue pelos vasos pulmonares em função da hipertensão arterial decorrente de fibrose e espessamento arteriolar determinada pela ação pneumotóxica da planta.

Experiments were performed to define the clinical and pathological picture of prolonged administration of the seeds of Crotalaria mucronata Desv. to cattle, in order to obtain additional information about this toxicosis. The ground seeds were administered orally to 9 bovines. Doses of 1g/kg/day, 2g/kg/day, 3g/kg/day each in one bovine, and 5g/kg/day in two of three bovines, given for 61- 63 days, did not cause poisoning. Doses of 5g/kg, in one bovine, 7.5g/kg in two bovines and 10g/kg in one bovine, given for 47-61 days, caused symptoms between 47 and 80 days after the first administration and caused death between 3 hours and 5 days after the onset of symptoms. The main clinical signs were positive venous pulse of the jugular vein, abdominal breath, tachycardia, loss of appetite, dry feces, sub-mandibular edema and weakness. Bovines that did not die, were slaughtered 8 or 9 months after first administration. At necropsy pulmonary paleness, hydropericardium, hydrothorax, hydroperitoneum, mesenteric edema, augmented hepatic consistency, discoloration of the liver, right cardiac ventricle dilatation and ruminal wall edema were seen. The main histological lesions were thickening of the alveolar walls and of the arterioles with narrowing of their lumen, and periarteriolar fibrosis, besides hepatic and cardiac lesions of minor importance. It is concluded, that the lesions caused by ingestion of the seeds of C. mucronata over a long period are caused by the difficulties of blood passage through the pulmonar vessels due to fibrosis and thickening of the arteriolar walls through the pneumotoxic action of the plant.

A case report of hepatic veno-occlusive disease after ingesting dainties.

Hepatic veno-occlusive disease (HVOD) is rarely encountered and easily misjudged as Budd-Chiari syndrome. It is often related to stem cell transplantation in recent years. We report a case of HVOD that is related to ingestion of some palatable local dishes. The diagnosis was confirmed by liver biopsy pathology with specific observation of inflammatory changes and fibrosis of venules intima, dilated sinusoids and central veins. Chronic diarrhea is unique for this case as a result of ingesting harmful stuffs. This case demonstrated that supervision and instruction of food recipe and traditional medicine are crucial, and prompt diagnosis, supportive care and specific treatment are essential to decreasing the morbidity and mortality of HVOD.