Proceedings Volume 2010 (format .pdf) - SimpBTH

evaluations of the specific physical-chemical indicators were done according to theFR X and the European Pharmacopeea [9-11].SubjectsFourteen healthy volunteers (7 males and 7 females) with a mean age of 30.2(±4.1) years old enrolled and signed an informed consent prior to their participationin the study. None of the volunteers have taken any medication for at least a week,did any exercise, or ate before the blood sample was drawn. All blood sampleswere drawn in the morning between 8 and 9 a.m.Human whole blood cultureBlood samples were drawn from fourteen healthy volunteers into sterilizedsodium heparin tubes and processed within 45 min. The blood as diluted with 1:10with RPMI 1640, supplemented with 2 mM glutamine, 100 U/ml penicillin and 100lg/ml streptomycin, without exogenous serum. To each well of the 6-well cultureplates, 2.0 ml of the diluted blood was added. To study cytokine modulation, 0.001,0.01, 0.1, 1, 10 or 100 ug of RZ/ml was added to each well, incubated for 24 h,followed by adding a mixture of Phytohemeagglutinin (PHA)+Lipopolysaccharide(LPS) in 40 µl volume to give a final concentrations of 5 and 1 µg/ml for PHA andLPS, respectively, and incubated in 5% CO2 at 37ºC for another 24 h. At the end ofincubation, blood was collected from wells into sterilized tubes and each well waswashed with 0.5 ml of RPMI to ensure removal of all well content. Thesupernatants were aliquoted and stored in sterilized tubes at -30ºC until assayed.Cytokine analysisCytokine level from supernatants was determined with preconfigured kitsusing the Luminex xMAP technologyStatistical analysisThe results were expressed as the means concentrationof each cytokine(±SD). Statistical comparisons between dosage groups were done using the pairedt-test and a one-way ANOVA, with P < 0.01 indicating significance.RESULTS AND DISCUSSIONStimulating or modulating the release of cytokines by immunomodulators orimmunostimulating agents is an attractive mode for treating several diseases. Ofthese agents, we found that the selective extract (table 1) obtained fromRosmarinus officinalis (RZ) can induce and modulate cytokines in dose-dependentmanner. RZ was found to reduce significantly (p < 0.001) IL-1β, TNF-α, and IL-6production from whole blood culture in a concentration dependent manner (fig. 1-3).241