Nehme Hala, Ayde Helena, El Obeid Dany, Sabatier Jean Marc, Fajloun Ziad
Bioactive Molecules Research Laboratory, Faculty of Sciences, Section II, Lebanese University, B.P. 90656, Jdeidet El-Matn 1202, Lebanon.
Faculty of Agriculture & Veterinary Sciences, Lebanese University, Dekwaneh, Beirut 2832, Lebanon.
Antibiotics (Basel). 2020 Nov 18;9(11):824. doi: 10.3390/antibiotics9110824.
Bacterial resistance has become a worrying problem for human health, especially since certain bacterial strains of () can cause very serious infections. Thus, the search for novel natural inhibitors with new bacterial targets would be crucial to overcome resistance to antibiotics. Here, we evaluate the inhibitory effects of bee venom (BV-) and of its two main components -melittin and phospholipase A (PLA)- on FF-ATPase enzyme, a crucial molecular target for the survival of these bacteria. Thus, we optimized a spectrophotometric method to evaluate the enzymatic activity by quantifying the released phosphate from ATP hydrolysis catalyzed by FF-ATPase. The protocol developed for inhibition assays of this enzyme was validated by two reference inhibitors, thymoquinone (IC = 57.5 μM) and quercetin (IC = 30 μM). Results showed that BV- has a dose-dependent inhibitory effect on FF-ATPase with 50% inhibition at 18.43 ± 0.92 μg/mL. Melittin inhibits this enzyme with IC = 9.03 ± 0.27 µM, emphasizing a more inhibitory effect than the two previous reference inhibitors adopted. Likewise, PLA inhibits FF-ATPase with a dose-dependent effect (50% inhibition at 2.11 ± 0.11 μg/mL) and its combination with melittin enhanced the inhibition extent of this enzyme. Crude venom and mainly melittin and PLA, inhibit FF-ATPase and could be considered as important candidates for combating resistant bacteria.
细菌耐药性已成为人类健康领域一个令人担忧的问题，尤其是某些（）细菌菌株可引起非常严重的感染。因此，寻找具有新细菌靶点的新型天然抑制剂对于克服抗生素耐药性至关重要。在此，我们评估了蜂毒（BV-）及其两种主要成分——蜂毒肽和磷脂酶A（PLA）对FF-ATP酶的抑制作用，该酶是这些细菌生存的关键分子靶点。因此，我们优化了一种分光光度法，通过定量FF-ATP酶催化ATP水解释放的磷酸盐来评估酶活性。用于该酶抑制试验的方案通过两种参考抑制剂百里醌（IC = 57.5 μM）和槲皮素（IC = 30 μM）进行了验证。结果表明，BV-对FF-ATP酶具有剂量依赖性抑制作用，在18.43±0.92 μg/mL时抑制率为50%。蜂毒肽以IC = 9.03±0.27 µM抑制该酶，其抑制作用比之前采用的两种参考抑制剂更强。同样，PLA对FF-ATP酶具有剂量依赖性抑制作用（在2.11±0.11 μg/mL时抑制率为50%），并且它与蜂毒肽的组合增强了该酶的抑制程度。粗毒液以及主要是蜂毒肽和PLA，抑制FF-ATP酶，可被视为对抗耐药细菌的重要候选物。
