Med. Weter. 70 (12), 781-785, 2014
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| Matysiak J., Dereziński P., Klupczyńska A., Cichocki M., Kokot Z. J. |
| Effect of bee venom on CYP1A2 activity |
| Bee venom is a complex mixture of substances of natural origin, whose therapeutic properties are used in many areas of medicine. Generally accepted procedures applied in the process of developing new drugs include tests that examine interactions between the new drug and the enzymes of the cytochrome P450 group (CYP450), which play a key role in the metabolism of xenobiotics and endogenous substances in mammals. The use of bee venom in the treatment of various diseases and in immunotherapy makes it necessary to test this material for its effect on the enzymes of the CYP450 group in order to prevent health-threatening interactions. The purpose of this paper was to investigate the effect of bee venom and its main component, melittin, on CYP1A2 enzyme activity. This enzyme plays an important role in the metabolism of many pharmaceuticals and toxins. This is the first study on the effect of bee venom on the activity of an enzyme of the CYP450 family. The CYP1A2/CEC High Throughput Inhibitor Screening Kit (BD Biosciences) was used in the study. The method was based on the measurement of fluorescence of the enzymatic reaction product (3-cyano-7- -hydroksycoumarin) formed by the action of the CYP1A2 on the substrate (3-cyano-7-ethoxycoumarin) in the presence of a potential inhibitor in various concentrations. Furafylline was used as a model inhibitor. Twenty samples of bee venom from different years and of different origin, as well as melittin, were analyzed. The tests were performed at 37°C in 96-well microplates with an Infinite M200 Pro (Tecan) microplate reader. Fluorescence measurement parameters were as follows: excitation – 410 nm, emission – 460 nm. On the basis on the results obtained, IC50 values were calculated, which are equal to the concentrations of particular inhibitors causing the inhibition of enzyme activity by 50%. The IC50 values against CYP1A2 for different samples of bee venom ranged from 0.13 μg/ml to 2.38 μg/ml (mean = 0.74 μg/ml). Comparison of the IC50 values for bee venom and furafylline (1.53 μg/ml) demonstrates potent inhibitor properties of bee venom against CYP1A2. The fact that IC50 values for different bee venom samples show a relatively high variability may be caused by composition differences between particular bee venom samples. The data obtained also indicate that melittin is a relatively weak inhibitor of CYP1A2 activity compared to bee venom (IC50 for melittin is 41.04 μg/ml). It can therefore be assumed that the inhibition of CYP1A2 by bee venom is caused by its other components. The results obtained highlight the problem of potential interactions between bee venom and therapy. |
| Key words: bee venom, melittin, cytochrome P450, CYP1A2 |