CAJ | 학술논문

目的探讨甘露聚糖结合凝集素(mannan-binding lectin,MBL)对白假丝酵母菌(Candida albicans)刺激的THP1/CD14细胞产生TNF-α和IL-8的影响.方法以不同浓度人MBL预处理THP1/CD14细胞2 h后,再用热灭活的酵母相C.albicans和/或菌丝相C. albicans刺激细胞24 h,收集培养上清,以ELISA从蛋白水平分析其TNF-α和IL-8的产生.收集细胞提取总RNA,以RT-PCR从转录水平评估TNF-α和IL-8的表达,Western blot分析NF-KB的细胞核移位.结果 ELISA检测发现,两种相态的C. albicans均可刺激各组细胞分泌TNF-α和IL-8,酵母相C.albicans刺激细胞产生细胞因子水平稍高;高浓度MBL(10～20 mg/L)均可抑制两种相态的C. albicans诱导细胞分泌TNF-α和IL-8,低浓度MBL(1 mg/L)则几乎无影响;RT-PCR分析亦显示,与相应只用C. albicans刺激的实验组相比,高浓度MBL(20 mg/L)对两种相态的C. albicans诱导的TNF-α、IL-B的mRNA表达均有不同程度的抑制作用;Western blot分析显示,高浓度MBL(20 mg/L)对两种相态的C. albicans诱导的NF-KB细胞核移位有显著的抑制作用.结论 MBL可抑制C.albicans诱导的THP1/CD14细胞产生TNF-α和IL-8,提示MBL能够在抗C.albicans 免疫中起调控作用.
목적 탐토감로취당결합응집소(mannan-binding lectin,MBL)대백가사효모균(Candida albicans)자격적THP1/CD14세포산생TNF-α화IL-8적영향.방법 이불동농도인MBL예처리THP1/CD14세포2 h후,재용열멸활적효모상C.albicans화/혹균사상C. albicans자격세포24 h,수집배양상청,이ELISA종단백수평분석기TNF-α화IL-8적산생.수집세포제취총RNA,이RT-PCR종전록수평평고TNF-α화IL-8적표체,Western blot분석NF-KB적세포핵이위.결과 ELISA검측발현,량충상태적C. albicans균가자격각조세포분비TNF-α화IL-8,효모상C.albicans자격세포산생세포인자수평초고;고농도MBL(10～20 mg/L)균가억제량충상태적C. albicans유도세포분비TNF-α화IL-8,저농도MBL(1 mg/L)칙궤호무영향;RT-PCR분석역현시,여상응지용C. albicans자격적실험조상비,고농도MBL(20 mg/L)대량충상태적C. albicans유도적TNF-α、IL-B적mRNA표체균유불동정도적억제작용;Western blot분석현시,고농도MBL(20 mg/L)대량충상태적C. albicans유도적NF-KB세포핵이위유현저적억제작용.결론 MBL가억제C.albicans유도적THP1/CD14세포산생TNF-α화IL-8,제시MBL능구재항C.albicans 면역중기조공작용.
Objective To investigate the effects of mannan-binding lectin (MBL) on IL-8 and TNF-α production induced by Candida albicans ( C. albicans) in human THP1/CD14 monocytes. Methods The THP1/CD14 cells were stimulated for 24 h with heat-inactivated yeast form or hyphal form cells of C. albicans strain at the indicated ratios after pretreated with human natural MBL at concentrations ranging from 1 to 20 mg/L for 2 h. The content of IL-8 and TNF-α in culture supernatants were detected by ELISA,and the levels of IL-8 and TNF-α mRNA expressions in these cells were determined by RT-PCR. Western blot was used to detect C. albicans-induced NF-κB translocation in THP1/CDI4 cells. Results ELISA showed that secretion of IL-8 and TNF-α from THP1/CD14 cells could be induced by both yeast cells and hyphal cells. Hyphal cells proved to be much less efficient than yeast cells in stimulating production of IL-8and TNF-α by THP1/CD14 cells. The productions of IL-8 and TNF-α by THP1/CD14 cells induced with C.albicans were profoundly inhibited by MBL at higher concentrations ( 10-20 mg/L) but not MBL at lower concentrations ( 1 mg/L). RT-PCR analysis also indicated that the mRNA expressions of IL-8 and TNF-αt in THP1/CD14 cells were decreased to various extents by MBL at higher concentration, compared to the corresponding THP1/CD14 cells stimulated with C. albicans only. Similarly, MBL at higher concentration ( 20mg/L) decreased the NF-κB translocation in THP1/CD14 cells. Conclusion MBL may inhibit IL-8 and TNF-α production induced by dimorphism C. albicans in THP1/CD14 cells, suggesting that MBL can play some roles on the regulation of C. albicans immune response.