Herrera Natalia, Herrera Carolina, Ortíz Isabel, Orozco Luz, Robledo Sara, Agudelo Diana, Echeverri Fernando
Grupo de Química Organica de Productos Naturales, Instituto de Química, Universidad de Antioquia, Calle 67 No. 53-10, Medellín, 050010, Colombia.
Grupo de Química Organica de Productos Naturales, Instituto de Química, Universidad de Antioquia, Calle 67 No. 53-10, Medellín, 050010, Colombia.
Toxicon. 2018 Nov;154:50-59. doi: 10.1016/j.toxicon.2018.09.011. Epub 2018 Sep 28.
The presence of cyanobacterial blooms and cyanotoxins in water presents a global problem due to the deterioration of ecosystems and the possibility of poisoning in human and animals. Microcystin LR is the most widely distributed cyanotoxin and liver cells are its main target. In the present study, HepG2 cells were used to determine DNA damage of three crude extracts of cyanobacterial blooms containing MC-LR, through comet assay. The results show that all extracts at a concentration of 500 μg mL caused low damage in hepatocytes exposed for 24 h, but produced total mortality even at low concentrations at 48 h. Moreover, balloons corresponding to cell apoptosis were found. Through HPLC/MS, MC-LR was detected in all samples of cyanobacterial blooms at concentrations of (5,65 μg ml) in sample 1, (1,24 μg ml) in sample 2 and (57,29 μg ml) in sample 3. In addition, in all samples high molecular weights peaks were detected, that may correspond to other microcystins. Besides, the cytotoxic effect of a cyanobacterial bloom and some of its chromatographic fractions from the crude extracts were evaluated in U-937, J774, Hela and Vero cell lines, using the enzymatic micromethod (MTT). The highest toxicity was detected in U-937 cells (LC = 29.7 μg mL) and Vero cells (LC = 39.7 μg mL-). Based on these results, it is important to remark that genotoxic and cytotoxicity assays are valuable methods to predict potential biological risks in waters contaminated with blooms of cyanobacteria, since chemical analysis can only describe the presence of cyanotoxins, but not their biological effects.
由于生态系统的恶化以及人类和动物中毒的可能性,水体中蓝藻水华和蓝藻毒素的存在是一个全球性问题。微囊藻毒素LR是分布最广的蓝藻毒素,肝细胞是其主要靶标。在本研究中,使用HepG2细胞通过彗星试验测定了三种含有MC-LR的蓝藻水华粗提物的DNA损伤。结果表明,所有浓度为500μg/mL的提取物在暴露24小时的肝细胞中造成的损伤较小,但即使在低浓度下,48小时时也会导致全部死亡。此外,还发现了与细胞凋亡相对应的空泡。通过HPLC/MS检测到,所有蓝藻水华样品中均含有MC-LR,样品1中的浓度为(5.65μg/mL),样品2中的浓度为(1.24μg/mL),样品3中的浓度为(57.29μg/mL)。此外,在所有样品中均检测到高分子量峰,这可能对应于其他微囊藻毒素。此外,使用酶促微量法(MTT)在U-937、J774、Hela和Vero细胞系中评估了蓝藻水华及其粗提物的一些色谱馏分的细胞毒性作用。在U-937细胞(LC=29.7μg/mL)和Vero细胞(LC=39.7μg/mL)中检测到最高毒性。基于这些结果,必须指出的是,遗传毒性和细胞毒性测定是预测受蓝藻水华污染水体中潜在生物风险的有价值方法,因为化学分析只能描述蓝藻毒素的存在,而不能描述其生物效应。