生长激素对人外周血来源NK细胞杀伤活性的影响及机制探讨

发布时间：2018-03-22 07:47

本文选题：生长激素　切入点：自然杀伤细胞　出处：《山东医药》2017年18期 　论文类型：期刊论文

【摘要】：目的观察生长激素(GH)对人外周血来源自然杀伤(NK)细胞杀伤活性的影响,并探讨其机制。方法选取10名健康志愿者,采用免疫磁珠法分离并培养其外周血NK细胞。将NK细胞分为A、B、C组,分别与终浓度0、5、20μg/mL的重组人GH共培养48 h。以人类髓性白血病K562细胞为靶细胞,NK细胞为效应细胞,按不同的效靶比(25∶1、50∶1、100∶1)混合,采用CCK-8法检测NK细胞对K562细胞的杀伤率;取各组培养48 h的NK细胞,分别采用流式细胞术和Real-time RT-PCR法检测NK细胞表面活化受体NKG2D蛋白及基因。结果三个不同效靶比下,B、C组NK细胞对K562细胞的杀伤率高于A组,且C组高于B组(P均0.01)。A、B、C组NKG2D阳性表达的NK细胞百分数分别为78.8%±2.8%、87.5%±3.1%、91.8%±3.3%,B、C组NKG2D阳性表达的NK细胞百分数高于A组(P均0.05)。A、B、C组NKG2D mRNA相对表达量分别为1±0.01、5±2.32、10±3.21,B、C组NKG2D mRNA相对表达量高于A组,且C组高于B组(P均0.05)。结论 GH可通过上调NK细胞NKG2D的表达,增强其杀伤活性。
[Abstract]:Objective to observe the effect of growth hormone (GH) on the cytotoxicity of NK cells from human peripheral blood and its mechanism. Methods Ten healthy volunteers were selected. NK cells in peripheral blood were isolated and cultured by immunomagnetic bead method. NK cells were divided into two groups: group C, co-cultured with recombinant human GH with final concentration of 0 ~ 5 渭 g/mL for 48 h. Human myeloid leukemia K562 cells were used as target cells and NK cells were used as effector cells. The cytotoxicity of NK cells to K562 cells was detected by CCK-8 assay, and the NK cells were cultured for 48 h. Flow cytometry and Real-time RT-PCR were used to detect the surface activated receptor (NKG2D) protein and gene of NK cells. Results the cytotoxicity of NK cells to K562 cells was higher in group C than in group A. The percentage of NKG2D positive NK cells in group C was higher than that in group B (78.8% 卤2.8% 卤3.1% 卤3.1% 卤3.1% 卤3.31%), respectively. The relative expression of NKG2D mRNA in group C was higher than that in group A (1 卤0.01 卤2.32 10 卤3.21). The relative expression of NKG2D mRNA in group C was higher than that in group A (P = 0.05 卤2.32 10 卤3.21), and the percentage of NK cells in group C was higher than that in group A (P = 0.05 卤2.32 10 卤3.21), and the percentage of NK cells in group C was higher than that in group A (P 0.05 卤2.32 10 卤3.21). Conclusion GH can enhance the cytotoxicity of NK cells by upregulating the expression of NKG2D.
【作者单位】：滨州医学院附属医院;
【基金】：国家自然科学基金资助项目(81171119) 滨州医学院科研计划课题项目(BY2015KJ28)
【分类号】：R392

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