CAJ | 학술논문

目的：比较不同实验条件对检测肺泡巨噬细胞吞噬功能的影响，为流式细胞术检测肺泡巨噬细胞吞噬功能筛选快速、灵敏、稳定的实验方法。方法：支气管肺泡灌洗获取肺泡巨噬细胞，将肺泡巨噬细胞分别与转红色荧光蛋白基因质粒细菌〔E.coliDH5α（pJZ402rfp)〕及未转质粒细菌（E.coliDH5α）孵育，并选择〔E.coliDH5α（pJZ402rfp)〕与肺泡巨噬细胞按不同的细菌与细胞比例(50：1、100：1、200：1)、孵育时间（30min、60min、120min、180min）及培养溶液（PBS、Hanks、无血清1640）进行孵育，流式细胞仪检测肺泡巨噬细胞的吞噬功能。结果：E.coliDH5α（pJZ402rfp)具有稳定的红色荧光，用E.coliDH5α（pJZ402rfp)检测肺泡巨噬细胞的吞噬功能比用E.coliDH5α获得更多的参数。肺泡巨噬细胞的吞噬率随着细菌与细胞比例的增大而增加，而吞噬指数在细菌与细胞比例为100：1时为最高（P<0.05）；肺泡巨噬细胞的吞噬率从30分钟到120分钟变化不大，但到180分钟吞噬率有明星的降低（P<0.05），吞噬指数则在60分钟时最高，有显著差异（P<0.05）；Hanks液组肺泡巨噬细胞的吞噬率及吞噬指数显著高于PBS组及1640组（P<0.05）。结论：在室温（260C）以肺泡巨噬细胞与E.coliDH5α（pJZ402rfp)比为1：100的Hanks液中孵育1小时，是流式细胞术检测肺泡巨噬细胞吞噬功能的快速、灵敏、稳定的实验方法。
목적：비교불동실험조건대검측폐포거서세포탄서공능적영향，위류식세포술검측폐포거서세포탄서공능사선쾌속、령민、은정적실험방법。방법：지기관폐포관세획취폐포거서세포，장폐포거서세포분별여전홍색형광단백기인질립세균〔E.coliDH5α（pJZ402rfp)〕급미전질립세균（E.coliDH5α）부육，병선택〔E.coliDH5α（pJZ402rfp)〕여폐포거서세포안불동적세균여세포비례(50：1、100：1、200：1)、부육시간（30min、60min、120min、180min）급배양용액（PBS、Hanks、무혈청1640）진행부육，류식세포의검측폐포거서세포적탄서공능。결과：E.coliDH5α（pJZ402rfp)구유은정적홍색형광，용E.coliDH5α（pJZ402rfp)검측폐포거서세포적탄서공능비용E.coliDH5α획득경다적삼수。폐포거서세포적탄서솔수착세균여세포비례적증대이증가，이탄서지수재세균여세포비례위100：1시위최고（P<0.05）；폐포거서세포적탄서솔종30분종도120분종변화불대，단도180분종탄서솔유명성적강저（P<0.05），탄서지수칙재60분종시최고，유현저차이（P<0.05）；Hanks액조폐포거서세포적탄서솔급탄서지수현저고우PBS조급1640조（P<0.05）。결론：재실온（260C）이폐포거서세포여E.coliDH5α（pJZ402rfp)비위1：100적Hanks액중부육1소시，시류식세포술검측폐포거서세포탄서공능적쾌속、령민、은정적실험방법。
ObjectiveThe purpose of this study was to establish a flow cytometric assay on the phagocytic function in rat alveo-lar macrophage (AM) Methods AM collected by bronchoalveolar lavage from rats phagocytes E.coliDH5α(pJZ402rfp) in different condition. The percentage of positive AM with associated bacteria was determined as phagocytic rate, and phagocytic index was de-tected as ration of fluorencence index (FI) of positive AM to FI of control AM, which were analyzed by flow cytometry. Results The red fluorescent protein of E.coliDH5α(pJZ402rfp) is bright and stabilized. There are more parameters obtained from AM chal-lenged with E.coliDH5α(pJZ402rfp) than E.coliDH5α. Phagocytic rate in AM increased with the ration of bacteria to AM. Howev-er, the highest phagocytic index was found in the ratio of 100:1 ( P<0.05) Phagocytic rate decreased markedly at 180min, com-pared with 30, 60, and 120 min. While the phagocytic index significantly increased at 60min ( P<0.05) Compared with PBS group and 1640 group, phagocytic rate and phagocytic index markedly increased in Hanks group ( P<0.05) Conclusions AM phago-cytes E.coliDH5α(pJZ402rfp) with the ration of 1:100 in the Hanks for 60 min at RT. We have developed a flow cytometric assay to detect phagocytic rate and phagocytic index in AM.