CAJ | 학술논문

目的：通过检测微囊藻毒素-LR(MC-LR)对BV-2细胞内一氧化氮(NO)和一氧化氮合酶(NOS)，以及培养上清液中的肿瘤坏死因子-α(TNF-α)和白介素-6(IL-6)表达的影响，为进一步探讨MC-LR引起中枢神经系统免疫反应奠定基础。方法：用MC-LR刺激BV-2细胞建立炎症模型，以四甲基噻唑蓝(MTT)法测定MC-LR对细胞的毒性作用；采用硝酸还原酶法检测BV-2细胞中NO的表达量；NOS分型法检测NOS的表达水平；酶联免疫吸附实验(ELISA)检测TNF-α和IL-6炎症因子的产生情况。结果：MC-LR染毒24 h后，染毒浓度在0～128μg/L范围内，随着剂量的增加，细胞活性呈下降趋势(r=-0.609，P<0.05)，其半数有效浓度(EC50)为22.49μg/L。当染毒浓度≥5.0μg/L时，细胞分泌的IL-6和NOS的含量明显增加，与对照组比较差异具有统计学意义(P<0.05)；10.0μg/L染毒后TNF-α和NO含量明显增加，与对照组比较差异具有统计学意义(P<0.05)。结论：MC-LR诱导BV-2细胞产生大量的炎性细胞因子NO、NOS、IL-6及TNF-α，引起神经系统的炎症反应。
목적：통과검측미낭조독소-LR(MC-LR)대BV-2세포내일양화담(NO)화일양화담합매(NOS)，이급배양상청액중적종류배사인자-α(TNF-α)화백개소-6(IL-6)표체적영향，위진일보탐토MC-LR인기중추신경계통면역반응전정기출。방법：용MC-LR자격BV-2세포건립염증모형，이사갑기새서람(MTT)법측정MC-LR대세포적독성작용；채용초산환원매법검측BV-2세포중NO적표체량；NOS분형법검측NOS적표체수평；매련면역흡부실험(ELISA)검측TNF-α화IL-6염증인자적산생정황。결과：MC-LR염독24 h후，염독농도재0～128μg/L범위내，수착제량적증가，세포활성정하강추세(r=-0.609，P<0.05)，기반수유효농도(EC50)위22.49μg/L。당염독농도≥5.0μg/L시，세포분비적IL-6화NOS적함량명현증가，여대조조비교차이구유통계학의의(P<0.05)；10.0μg/L염독후TNF-α화NO함량명현증가，여대조조비교차이구유통계학의의(P<0.05)。결론：MC-LR유도BV-2세포산생대량적염성세포인자NO、NOS、IL-6급TNF-α，인기신경계통적염증반응。
OBJECTIVE:To investigate the effects ofmycrocystin-LR(MC-LR) on the neurotoxic mechanisms of the central nervous system,the contents of nitric oxide (NO),nitric oxide synthase (NOS),tumor necrosis factor-α(TNF-α),and interleukin- 6 (IL-6) in BV-2 microglial cells were detected in our study.METHODS:With the methyl thiazolyl tetrazolium (MTT) assay,the cytotoxicity of the inflammatory model in BV-2 cells was performed after exposure to MC-LR. The levels of NO and NOS were measured by the nitric acid reductase assay and nitric oxide synthase assay,respectively. The contents of TNF-α and IL-6 were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS:After exposure to MC-LR for 24 h,theviability in BV-2 cells was decreased with the increase of MC-LR concentration within the range of 0-28μg/L (r=-0.609,P<0.05),and the median effective concentration (EC50) was 22.49μg/L. Besides,the levels of IL-6 and NOS in BV-2 cells were increased at the concentration of 5-10μg/L MC-LR,and the levels of TNF-α and NO were increased at the concentration of 10μg/L MC-LR. Compared withcontrol group,those differences above mentioned were statistically significant (P<0.05).CONCLUSION:MC-LR induced inflammatory response in BV-2 microglial cells by increasesin p roductionof NO,NOS,IL-6 and TNF-α.