Ficus glumosa Del. reduces phenylhydrazine-induced hemolytic anaemia and hepatic damage in Wistar rats.

OBJECTIVES: Anemia is a direct or indirect consequence of oxidative stress via free radicals on erythrocytes and subsequently on other tissues like liver. Ficus glumosa constitute a rich pharmacologically compound that can prevent or repair oxidative damage. Therefore, this study seeks to evaluate the effect of F. glumosa on phenylhydrazine-induced hemolytic anemia and hepatic damage in rats. METHODS: Twenty-four (24) albino Wistar rats were assigned to four (4) experimental groups (n=6) as follows: Group I (non-anemic control) and Group 2 (anemic control) received normal saline, while Group III and IV (test groups) 200 and 400 mg/kg of aqueous leaf extract of F. glumosa (ALEFG), respectively. All the groups were treated orally (via a cannula) for seven consecutive days. Intraperitoneal (IP) injection of phenylhydrazine (PHZ) at 40 mg/kg for two consecutive days induced hemolytic anemia in group II to IV before treatment. Rats of all groups were anaesthetized and sacrificed 24 h after the last treatment. Blood and liver samples were collected for some hematological indices, liver function test, antioxidant parameter and histological analysis. RESULTS: The LD50 of ALEFG was assessed orally in rats and found to be above 5,000 mg/kg body weight. Significant (p<0.05) decreases in the level of red blood cell (RBC), hemoglobin (HGB) concentrations and packed cell volume (PCV) by 50% after 2 days of PHZ induction, were attenuated by more than 50% after 7 days administration of ALEFG at 200 and 400 mg/kg. The percentage change in body weight increased significantly (p<0.05) after 7 days post PHZ-induced anemia, but those that received oral administration of ALEFG (at 200 and 400 mg/kg) for 7 days increased significantly (p<0.05) by more than 2%, dose-dependently compared to anemic untreated group. Increased level of serum ALT, AST, ALP and GGT in PHZ-induced anemic animals, were significantly (p<0.05) attenuated in the groups that received oral administration of ALEFG (at 200 and 400 mg/kg) for 7 days. Decreased level of catalase (CAT) and superoxide dismutase (SOD) activities with concomitant increase in malondialdehyde (MDA) content from PHZ-induced untreated group, were significantly (p<0.05) mitigated in the rats that received oral administration of ALEFG (at 200 and 400 mg/kg) for 7 days. Histopathological analysis showed that ALEFG could remarkably though not completely mitigated PHZ-induced hepatic damage. CONCLUSIONS: Our data suggests that the leaves of F. glumosa contain important antioxidant(s) that could effectively reduce hemolytic anemia and hepatic damage, especially during phenylhydrazine-induced toxicity.

Phenylhydrazine administration accelerates the development of experimental cerebral malaria.

Phenylhydrazine (PHZ) treatment is generally used to enhance parasitemia in infected mice models. Transient reticulocytosis is commonly observed in iron-deficient anemic hosts after treatment with iron supplementation, and is also associated with short-term hemolysis caused by PHZ treatment. In this study, we investigated the relationship between reticulocytosis and cerebral malaria (CM) in a murine model induced by PHZ administration before Plasmodium berghei ANKA (PbA) infection. Mortality and parasitemia were checked daily. Pro-inflammatory cytokines and IL-10 were quantified by ELISA. The expression of CXCL9, CXCL10, CCL5, and CXCR3 mRNAs was determined by real-time PCR. Brain sequestration of CD4(+) and CD8(+) T cells and populations of splenic Th1 CD4(+) T cells, dendritic cells (DCs), CD11b(+) Gr1(+) cells, and regulatory T cells (Tregs) were assessed by FACS. PHZ administration dramatically increased parasitemia from day 3 to day 5 post infection (p.i.) compared with the untreated control infected mice group; also, CM developed at day 5 p.i., compared with day 7 p.i. in untreated control infected mice, as well as significantly decreased blood-brain barrier function (P < 0.001). PHZ administration during PbA infection significantly increased the expression of CXCL9 (P <0.05) and VCAM-1 (P <0.001) in the brain, increased the expression of CXCL10, CCL5 and CXCR3, and significantly increased the recruitment of CD4(+) and CD8(+) T cells (P <0.001 and P <0.01, respectively) as well as CD11b(+) Gr1(+) cells to the brain. In addition, PHZ administration significantly increased the numbers of IL-12-secreting DCs at days 3 and 5 p.i. compared to those of untreated control infected mice (P <0.001 and P <0.01, respectively). Consequently, the activation of CD4(+) T cells, especially the expansion of the Th1 subset (P <0.05), was significantly and dramatically enhanced and was accompanied by marked increases in the production of protein and/or mRNA of the Th1-type pro-inflammatory mediators, IFN-γ and TNF-α (P <0.01 for both for protein; P <0.05 for TNF-α mRNA). Our results suggest that, compared to healthy individuals, people suffering from reticulocytosis may be more susceptible to severe malaria infection in malaria endemic areas. This has implications for the most appropriate selection of treatment, which may also cause reticulocytosis in patients living in such areas.

[Effect and its mechanism of Coptis chinensis on oxidative hemolysis of erythrocytes in mice induced by acetylphenylhydrazine].

OBJECTIVE: To investigate the effect of Coptis chinensis on oxidative hemolysis of erythrocytes in mice. METHOD: Acetylphenyhydrazine (APH)-induced oxidative hemolysis of erythrocytes in mice were used. The contents of free hemoglobin of blood plasma, indirect bilirubin of serum, reticulocytes of blood, and malondialdehyde (MDA) of erythrocytes were measured. The activity of superoxide dismutase (SOD), reduced glutathione hormone (GSH), and glucose-6-phosp hate dehydrogenase (G-6-PD) of erythrocytes were also determined and the total-antioxygen capability (T-AOC) of blood was analyzed. RESULT: The levels or amount of free hemoglobin of blood plasma, indirect bilirubin of serum, reticulocytes of blood and MDA of erythrocytes were higher in APH (0.03 g x kg(-1))-induced mice than normal mice. The activity or content of SOD, GSH and G-6-PD was lower in APH-induced mice than in normal mice. Primaquine (0.058 g x kg(-1)) could aggravated the degree of elevated hemolysis of erythrocytes in APH-induced mice. C. chinensis (0.6 g x kg(-1) could deprssed significantly the elevated levels of indirect bilirubin in serum. The levels of free hemoglobin of blood plasma, indirect bilirubin of serum, reticulocytes of blood, the production of SOD and GSH and T-AOC were also decressed by C. chinensis (0.6 g x kg(-1)). CONCLUSION: C. chinensis suppressed t he degree of hemolysis of erythrocytes in APH-induced mice due to the suppression of the production of lipid peroxidation and increasing of the activity of antioxidase of erythrocytes.

Inadequate treatment of excessive erythrocytosis / Tratamiento inadecuado en la eritrocitosis excesiva

Los pacientes con el mal de montaña crónico o eritrocitosis excesiva (EE) son residentes de la altura (3600 m), con mayor o igual 6.5 x 10 a la sexta glóbulos rojos (GR) que presentan cianosis.Esto ocasiona problemas estéticos y psicológicos en su vida ya que las demás personas creen que son alcohólicos. Cuando hay aumento de los GR, ellos buscan una cura milagrosa. De acuerdo a los conceptos evolutivos de la EE, los tratamientos han incluído; sanguijuelas, radioterapia de la médula ósea mediante administración de substancias radiactivas como el fósforo, y más recientemente, flebotomías, infusiuones de té, tabletas de ajo y la más peligrosa la administración de la fenilhidrazina, agente citotóxico prohibido. Encontramos que la mayoría de los pacientes con EE tienen placas radiográficas de tórax anormales. El concepto de los tratamientos es el de disminuir los GR. Sin embargo, la fenilhidrazina es tóxica para la médula ósea, el hígado y otros tejidos, cambiando el color de la piel de cianótica a icterica. Las conjuntivas se tornan ictéricas y la harina de café oscura. Una vez iniciado el tratamiento, la sangre de los pacientes es analizada periódicamente y el recuento de GR disminuyue, con lo que quedan satisfechos. Sin embargo, este medicamento tóxico puede producir la muerte. Al reducir los GR, el contenido arterial de oxígeno (CaO2) en la sangre disminuye. Las pruebas ergométricas en estos pacientes durante el tratamiento producen gran débito de oxígeno. En el paciente descrito, en el 4to nivel del protocolo de Bruce,m el dolor intenso de ambasd pantorrillas se hizo intolerable y requirió oxígeno post ejercicio. Al interrumpirse la fenilhidrazina, el, CaO2 retorna a niveles normales en aproximadamente 60 días, con una elevación de los GR por encima de los valores iniciales, y mejoría de la capacidad de ejercicio. Este y muchos otros casos nos llevan a creer que la EE es un mecanismo de compensación de la enfermedad pulmonar en la altura y que la cantidad de GR no debe ser disminuída.