摘要
目的在黄芪多糖(APS)作用于骨髓单个核细胞(BMNC)来源的树突状细胞(DC)成熟过程中,研究其是否可以增加树突状细胞来源的外泌体(Dex)表面功能分子的表达水平。方法分离急性髓细胞白血病完全缓解期(AML-CR)患者BMNC,采用含有重组人粒细胞巨噬细胞集落刺激因子(r HuGM-CSF)和重组人白细胞介素4(rhuIL-4)的RPMI 1640完全培养基培养在37℃5%CO2细胞培养箱中培养至第7天。阳性对照组加常规量的脂多糖(LPS),实验组用APS 50、100、200 g/L刺激DC,阴性对照组加相同体积的培养基,培养36小时后,分级分离法提取DC培养上清液中Dex,在透射电镜下观察Dex的形态,流式细胞仪检测Dex表面分子人白细胞抗原DR、CD80、CD86的表达水平。并对DC进行形态学和表面分子鉴定。结果APS组与对照组所诱生的Dex的形态学差异无统计学意义。中剂量APS组中Dex表面分子(CD80、CD86、HLA-DR)表达量较其他各组显著增高(P<0.05)。结论APS可以增加急性髓细胞白血病患者BMNC来源的DC诱生的Dex表面分子的表达量。
Objective To explore the augmentation of astragulus polysaccharide（APS） on surface molecular expression of exosomes（Dex） derived from dentritie cells（DCs） by bone marrow mononuclear cells（BMNCs） in vitro. Methods BMNCs from acute mylocytic leukemia complete remission（AML-CR） patients were cultured in RPMI1640 medium containing recombinant human granulocyte-macrophage colony stimulating factor（GM-CSF）/recombinant human interleukin 4（IL-4） placed in 5% COs air at 37 ℃ incubator. After 7 days, positive control group was affected by a conventional close of lipopolysaccharide（LPS）,experimental groups were stimulated by APS at 50,100,200 g/L dose, negative control group was treated by RPMI1640 medium of an equal volume. After 36 hours of stimulation,exosomes were extracted from the supernatant of DCs culture by repeated ultraeentrefugation, the morpbologic features of exosomes（Exo） were observed by transmission electron microscope and the exosomal phenotype was detected by flow cytometry. Besides,the DCs were identified by Wright staining and flow cytometry. Results There was no statistical significance about exosomal morphology among different groups. Compared with control groups, APS groups increased the expression of HLA-DR, CD80, CD86 on the Dex （P 〈 0.05）. Conclusion APS could enhance the monocular expression on Dex in AML-CR patients.
出处
《临床荟萃》
CAS
2009年第12期1029-1032,F0003,共5页
Clinical Focus
关键词
白血病
粒细胞
急性
单核细胞
膜糖蛋白类
培养基
leukemia, myelocytic, acute
monocytes
membrane glycoproteins
culture media
