Antioxidant, genotoxic, antigenotoxic, and antineoplastic activities of apitoxin produced by Apis mellifera in Northeast, Brazil

ABSTRACT: The objective was to evaluate the in vitro antioxidant, genotoxic, antigenotoxic, and antineoplastic activities of apitoxin produced by the bee Apis mellifera. The antioxidant activity of the apitoxin solution was evaluated using the DPPH (2,2-diphenyl-1-picrilhydrazyl) method. Genotoxic potential of apitoxin was analyzed by comparing the mean DNA damage indices (idDNA) of L929 strain fibroblasts exposed to hydrogen peroxide (H2O2 - genotoxic substance), distilled water, or apitoxin. The antigenotoxic effect of apitoxin was analyzed by assessing the percentage decrease in H2O2-induced genotoxicity in L929 fibroblasts co-treated with three concentrations of the aqueous apitoxin solution and subjected to comet assay. In vitro antineoplastic activity in human tumor cell lines of prostate adenocarcinoma (PC3), hepatocellular carcinoma (HEPGE2), melanoma (MAD-MB435), and astrocytoma (SNB19), were verified by MTT [3- (4) bromide colorimetric method, 5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium]. Apitoxin had no genotoxic effect on L929 cells at concentrations of 30, 10, and 5 µg/mL after 24 hours of exposure. This effect was only evident at 50 µg/mL. Apitoxin promoted a significant reduction in DNA damage index (idDNA) at all concentrations tested. At 30 µg/mL, apitoxin attenuated the genotoxic effects induced by H2O2. Apitoxin also demonstrated in vitro antineoplastic potential, since the cytotoxic effect was observed at concentrations of 50 µg/mL and 25 µg/mL, with significant reduction in viability percentage of PC3 tumor cell lines, HEPGE2, MAD-MB435, and SNB19. The high antioxidant activity associated with the absence of genotoxic effect and the genoprotective and antineoplastic effect demonstrated by apitoxin here provide indications of apitoxin’s therapeutic potential.

摘要：本研究旨在评估西方蜜蜂（Apis mellifera）所产蜂毒（apitoxin）的体外抗氧化、遗传毒性、抗遗传毒性及抗肿瘤活性。采用二苯基苦基苯肼（DPPH，2,2-diphenyl-1-picrylhydrazyl）法评估蜂毒溶液的抗氧化活性。通过对比暴露于过氧化氢（H₂O₂，遗传毒性物质）、蒸馏水或蜂毒的L929成纤维细胞株的平均DNA损伤指数（idDNA），分析蜂毒的遗传毒性潜力。通过彗星实验评估经三种浓度蜂毒水溶液共处理且受H₂O₂诱导遗传毒性的L929成纤维细胞中H₂O₂诱导的遗传毒性百分比降幅，以此分析蜂毒的抗遗传毒性效应。采用MTT[3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐]比色法，检测前列腺腺癌（PC3）、肝细胞癌（HEPGE2）、黑色素瘤（MAD-MB435）及星形细胞瘤（SNB19）人源肿瘤细胞系的体外抗肿瘤活性。暴露24小时后，浓度为30、10及5 μg/mL的蜂毒对L929细胞无遗传毒性效应，仅在50 μg/mL浓度下显现该效应。蜂毒可使所有受试浓度下的DNA损伤指数（idDNA）显著降低。在30 μg/mL浓度下，蜂毒可减弱H₂O₂诱导的遗传毒性效应。蜂毒亦展现出体外抗肿瘤潜力：在50 μg/mL及25 μg/mL浓度下可观察到细胞毒效应，显著降低PC3、HEPGE2、MAD-MB435及SNB19肿瘤细胞系的细胞存活率百分比。本研究中蜂毒所展现出的高抗氧化活性、无遗传毒性特性，以及遗传保护与抗肿瘤效应，均提示蜂毒具备治疗应用潜力。