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为了探讨细胞因子在三氧化二砷诱发APL细胞白细胞升高的作用 ,首先按照FAB细胞形态学和细胞遗传学标准选择APL患者。常规Ficoll密度梯度法分离骨髓和PBMC ,体外培养 1h去除贴壁细胞。体外IL 1β、IL 6、IL 8、TNF α和G CSF分泌水平采用ELISA方法进行测定 ,NBT还原实验用以检测APL细胞分化状态。直接细胞计数法和MTT法观察细胞增殖变化。研究结果证实体外 10 6M或体内以 10mg/d三氧化二砷治疗 96h后 ,APL细胞分泌IL 1β和G CSF平均水平升高 (P <0 0 5 ) ,IL 6和IL 8水平降低 (P <0 0 5 ) ,TNF α水平无明显变化 (P >0 0 5 )。进一步分析结果表明 ,体外APL细胞的增值比率与IL 1β或G CSF分泌升高比率呈现正相关 ,检测到IL 1β或G CSF患者的细胞数量增加比率高于未检测到IL 1β或G CSF组 ,说明IL 1β或G CSF分泌增加在三氧化二砷诱导后的APL细胞增殖中起着一定作用
In order to investigate the role of cytokines in arsenic trioxide-induced leukocyte proliferation in APL cells, APL patients were first selected according to FAB cell morphology and cytogenetic criteria. Bone marrow and PBMCs were isolated by routine Ficoll density gradient method and cultured for 1 hour to remove adherent cells. The levels of IL 1β, IL 6, IL 8, TNF α and G CSF secreted in vitro were determined by ELISA. The NBT reduction assay was used to detect the differentiation status of APL cells. The cell proliferation was observed by direct cell counting and MTT assay. The results showed that the average levels of IL-1β and G-CSF secreted by APL cells increased (P <0 05) and the levels of IL 6 and IL 8 decreased (P 0 05) in vitro at 10 6M or 10 mg / d after arsenic trioxide treatment ), TNFα level had no significant change (P> 0.05). Further analysis showed that in vitro APL cell proliferation rate and IL 1β or G CSF secretion increased positive correlation was detected IL 1β or G CSF in patients with increased cell population was higher than the rate of undetected IL 1β or G CSF group, Indicating that increased IL 1β or G CSF secretion plays a role in the proliferation of APL cells induced by arsenic trioxide