EFFECTS OF HESPERIDINE METHYLCHALCONE AND VITAMIN C AND THEIR COMBINATION WITH RUSCUS EXTRACT AND OF MICRONIZED DIOSMINE ON ISCHEMIA FOLLOWED BY REPERFUSION
Authors: Fatima Zely G.A. Cyrino and Eliete Bouskela
Affiliation: Laboratory for Clinical and Experimental Research on Vascular Biology (BioVasc), Biomedical Center, State University of Rio de Janeiro, Rio de Janeiro – RJ - Brazil
OBJECTIVES:In the present study, we have investigated theeffects of drugs commonly used to treat chronic venous diseases, namely hesperidinemethylchalcone (HMC), vitamin C and their combination with Ruscusextract, micronized diosmine and their respective vehicles on macromolecular permeability increase and leukocyte-endothelium interaction induced by 30 minutes(min) ischemia followed by reperfusion.
MATERIAL AND METHODS:Investigated drugs were given by oral route (twice a day, at 8:00 AM and 5:00 PM, every day during two weeks) in 3 different doses to 171 male hamsters (Mesocricetus auratus). WhenRuscus, HMC and VitC were given in combination it was according to the proportions found in Cyclo 3 Fort® (Ruscusextract 150 mg, HMC 150 mg of sterolicheterosides and ascorbic acid 100 mg).Anesthesia was induced by i.p. injection of ketamine/xylazineand maintained with α-chloraloseiv.Local ischemia of the cheek pouch was obtained by a cuff, made of thin latex tubing,mounted around the neck of the everted pouch where it leaves the mouth of the hamster. For macromolecular permeability measurements, 30 min after completion of the preparative procedure, FITC-dextran (MW 150,000 Dalton), was given iv, in a dose of 25 mg/100 g body weight. For leukocyte-endothelium interaction, prior to ischemia and 30 min after the onset of reperfusion, leukocytes were stained with an i.v. infusion of rhodamine G (5 mg/100 g/5 min).Fluorescent leukocytes adhering to the endothelium and rolling closer to the venular wall were quantified in UV-light microscope in an area of 2 by 2 mm.
RESULTS:(1) Macromolecular permeability increase: vitC, HMC, the mixture (vitC + HMC + Ruscusextract) and micronized diosmine decreased it proportionally to the oral dose given to the animals, compared to their respective placebos. (2) Number of adhering leukocytes: VitC significantly decreased it, proportionally to the oral dose given to the animals;at the lowest dose, HMC did not have any significant effect compared to its placebo but at all otherdoses tested, it showed a significant decrease;at all doses tested, the mixture of vitC + HMC + Ruscusextract significantly decreased itand in several instances it was significantly more active than micronized diosmine;micronized diosmine significantly decreased it in all doses tested but the effect was not proportional to the oral dose given to the animals.(3) Number of rolling leukocytes: at the lowest and intermediary doses, vitC did not have any significant effect compared to its placebo but at the highest dose, it elicited a decrease; at the lowest dose, HMCdid not have any significant effect compared to its placebo but at all other doses tested, it showed a significant decrease, in several instances more pronounced than micronized diosmine; at thelowest and intermediary doses, the mixture of vitC + HMC + Ruscusextract did not have any significant effect compared to its placebo, butat the highest dose, it decreased it and was significantly more active than micronized diosmine; micronized diosminedecreased it in all doses tested but the effect was not proportional to the oral dose given to the animals.
CONCLUSION: CVD patients have ischemia in the lower limbs due to venous stasis and it has been shown that active extracellular superoxide dismutase (EC-SOD) significantly reduced the postischemic permeability increase when present during the ischemic period. Our results strongly suggest that the combination of Ruscusextract, HMC and VitChas antioxidant properties.