CAJ | 학술논문

本研究旨在探讨活性氧（ ROS）清除剂N－乙酰－L－半胱氨酸（ NAC）能否抑制滴虫排泄分泌物（ Tv ESP）诱导的人子宫颈癌SiHa细胞凋亡。体外培养阴道毛滴虫并制备Tv ESP，分别用100μg／mL Tv ESP和PBS处理SiHa细胞， Tv ESP处理SiHa细胞30 min前加入NAC，进行预处理。使用CellTiter 96 AQueous单溶液细胞增殖检测试剂盒检测细胞存活率；利用2′，7′－二氯荧光黄双乙酸盐（ DCFH－DA）进行荧光探针标记测定胞内ROS水平；免疫印迹法检测cleaved caspase－3和PARP－1蛋白表达。结果显示， NAC预处理可降低Tv ESP的细胞毒性作用。 Tv ESP作用SiHa细胞16 h后，细胞内ROS水平升高，但经过NAC预处理后胞内ROS生成显著降低。此外， NAC预处理后能显著下调Tv ESP诱导的cleaved caspase－3和cleaved PARP－1的表达水平。
본연구지재탐토활성양（ ROS）청제제N－을선－L－반광안산（ NAC）능부억제적충배설분비물（ Tv ESP）유도적인자궁경암SiHa세포조망。체외배양음도모적충병제비Tv ESP，분별용100μg／mL Tv ESP화PBS처리SiHa세포， Tv ESP처리SiHa세포30 min전가입NAC，진행예처리。사용CellTiter 96 AQueous단용액세포증식검측시제합검측세포존활솔；이용2′，7′－이록형광황쌍을산염（ DCFH－DA）진행형광탐침표기측정포내ROS수평；면역인적법검측cleaved caspase－3화PARP－1단백표체。결과현시， NAC예처리가강저Tv ESP적세포독성작용。 Tv ESP작용SiHa세포16 h후，세포내ROS수평승고，단경과NAC예처리후포내ROS생성현저강저。차외， NAC예처리후능현저하조Tv ESP유도적cleaved caspase－3화cleaved PARP－1적표체수평。
This study intends to investigate whether the reactive oxygen species ( ROS ) scavenger N-aeetyl-L-cysteine ( NAC ) can inhibit Trichomonas vaginalis excretory-secretory products ( Tv ESP ) induced apoptosis of human cervical cancer ( SiHa) cells.T.vaginalis were in vitro cultured and Tv ESP were prepared.SiHa cells were treated with 100 μg/mL Tv ESP, PBS and NAC was pretreated with SiHa cells 30 min prior to exposure to 100 μg/mL Tv ESP.The cell viability was evaluated using CellTiter 96 AQueous One Solution Cell Proliferation Assay kit, and the intercellular ROS level was measured by fluorescent probe 2′, 7′-dichlorfluorescein-diacetate ( DCFH-DA ) .Cleaved caspase-3 and PARP-1 protein levels were detected by Western blot.Our results demonstrate that pretreatment of NAC reduced Tv ESP induced SiHa cell cytotoxicity.Intracellular ROS levels were highly induced after exposure to 100 μg/mL Tv ESP for 16 h, however, intracellular ROS levels were significantly reduced by pretreatment of NAC.In addition, pretreatment of NAC, significantly reduced cleaved caspase-3 and cleaved PARP-1 protein levels induced by Tv ESP.