摘要
目的 探究低浓度丁酸和白细胞介素5(IL-5)诱导人嗜酸性粒细胞白血病细胞系(EoL-1)分化为嗜酸性粒细胞的最优时间。方法 使用50μM丁酸和1 ng/ml IL-5诱导EoL-1细胞分化,分别在第0、3、6、9 d时,通过倒置显微镜观察细胞生长状态,并拍照记录。通过实时定量荧光聚合酶链式反应(Q-PCR)技术检测第0、3、5、6、9 d时,嗜酸性粒细胞相关因子的相对表达情况。HE染色观察细胞核形态变化及胞浆中嗜酸性颗粒蛋白染色情况。结果 低浓度丁酸和IL-5刺激后,光镜下可见随着刺激天数增加,细胞增殖速度降低,成团生长减少,细胞数量降低。Q-PCR结果显示,嗜酸性粒细胞相关因子IL-5、IL-10、IL-13、IL-17A、干扰素γ(IFN-γ)、基质金属蛋白酶9(MMP-9)、趋化因子26(CCL-26)的表达水平在第5天、第6天高于其他天数。HE染色结果显示,刺激6 d时,细胞核由椭圆形变为双叶核,细胞质中嗜酸性颗粒增多,胞浆被染为红色。结论 低浓度丁酸和IL-5诱导EoL-1细胞分化为嗜酸性粒细胞,6 d左右为最优时间。
OBJECTIVE To investigate the optimal time for low concentrations of butyric acid and interleukin 5(IL-5) to induce differentiation into eosinophils in human eosinophilic leukemia cell line(EoL-1).METHODS EoL-1 cell differentiation was induced by 50 μM butyric acid and 1 ng/ml IL-5,and the cell growth status was observed by inverted microscope on days 0,3,6 and 9,respectively,and photographed and recorded.The relative expression of eosinophil-associated factors at days 0,3,5,6,and 9 was detected by real-time quantitative fluorescence polymerase chain reaction(Q-PCR).Observe the morphological changes of the nucleus and the staining of eosinophil granule proteins in the cytoplasm by hematoxylin-eosin staining(HE).RESULTS After stimulation with low concentration of butyric acid and IL-5,it was visible under light microscopy that with the increase of stimulation days,the rate of cell proliferation decreased,the growth of clumps decreased,and the number of cells decreased.Q-PCR results showed eosinophil-associated factor IL-5,IL-10,IL-13,IL-17A,interferon-γ(IFN-γ),matrix metalloproteinase 9(MMP-9) and C-C motif chemokine ligand 26(CCL26) were higher on the 5th and 6th days,The HE staining results showed that after 6 days of stimulation,the nucleus changed from elliptical to double lobed,the number of eosinophilic particles in the cytoplasm increased,and the cytoplasm was stained red.CONCLUSION Low concentrations of butyric acid and IL-5 induce differentiation of EoL-1 cells into eosinophils for about 6 days.
作者
王晶
闫冰
任益民
王成硕
张罗
WANG Jing;YAN Bing;REN Yimin;WANG Chengshuo;ZHANG Luo(Beijing Tongren Hospital,Capital Medical University,Beijing,100730,China;Beijing Institute of Otolaryngology,Beijing Key Laboratory of Nasal diseases,Beijing,100005,China;Department of Otolaryngology Head and Neck Surgery,Beijing Tongren Hospital,Capital Medical University,Beijing,100005,China;Department of Allergy,Beijing Tongren Hospital,Capital Medical University,Beijing,100005,China;Research Unit of Diagnosis and Treatment of Chronic Nasal Diseases,Chinese Academy of Medical Sciences,Beijing,100005,China)
出处
《中国耳鼻咽喉头颈外科》
CSCD
2023年第7期443-447,共5页
Chinese Archives of Otolaryngology-Head and Neck Surgery
基金
国家重点研发计划(2022YFC2504100)
国家自然科学基金(82025010、81630023、82171108、81900917、81870698)
东城区优秀人才计划(2022-dchrcpyzz-3)。
