摘要
目的探讨吸入性麻醉药七氟醚对脐血来源未成熟树突状细胞成熟及成熟树突状细胞(DCs)活力的影响。方法密度梯度离心法分离脐血来源单核细胞,并随机分为3组,即常规培养组(C组)作为对照组、未成熟DC组(I组)和成熟DC组(M组)。C组加入重组人粒细胞—单核细胞集落刺激因子(rhGM-CSF)50 ng/ml和重组人白介素-4(rhIL-4)10 ng/ml共培养12d,然后加入肿瘤坏死因子(TNF-α)50ng/ml,持续培养至第14天。I组、M组分别于第6、13天用含2.5%七氟醚的混合气体处理,余同C组。倒置显微镜及扫描电镜下观察细胞形态,流式细胞技术(FCM)检测细胞表面免疫表型表达(CD80/CD86、CD83/HLA-DR),MTT法检测DC刺激T细胞增殖活力,ELISA法检测上清液中IL-12水平。结果 (1)DCs形态学:C组及M组细胞表面有大量较长突起,面I组细胞表面树突状突起短,分支少。(2)CD80/CD86、CD83/HLA-DR表达率:C组、M组、I组分别为(36.9±2.8)%和(12.9±2.4)%、(39.4±2.1)%和(14.4±3.0)%、(19.6±2.6)%和(7.1±1.4)%,I组与C组比较,差异有统计学意义(P<0.05),而M组与C组比较,差异无统计学意义(P>0.05)。(3)DCs刺激T细胞增殖活力:C组、M组、I组分别为1.44±0.23、1.37±0.10和0.56±0.12,I组与C组比较,差异有统计学意义(P<0.05),而M组与C组比较,差异无统计学意义(P>0.05)。(4)DCs分泌IL-12水平:C组、M组、I组分别为(958±321)pg/ml、(982±408)pg/ml和(377±312)pg/ml,I组与C组比较,差异有统计学意义(P<0.05),而M组与C组比较,差异无统计学意义(P>0.05)。结论七氟醚可抑制脐血来源树突状细胞成熟,但对成熟树突状细胞活力影响不明显。
Objective To study the effects of sevoflurane on the growth of dendritic cells and the vatilis of mature dendritic cells（DCs）from umbilical cord blood,through expousure to sevoflurane.Methods Cord blood mononuclear cytes （CBMC） were separated by centrifugal in density gradient.The cells were randomly allocated to three groups：the normal cultured group（group C）,the immature type group（group I） and the mature type group（group M）.Group C was cultured in the presence of rhGM-CSF（50 ng/ml） and rhIL4（10 ng/ml） for 14 days and rhTNF-α（50 ng/ml） was added into till the 12th day.The cells of Group I and Group M were exposured to 2.5%sevoflurane till the 6th and 13th day for 2h,respectively. The growth and morphology of DCs were identified by inverted optical microscope and scanning electron microscope.The phenotypes of 14 day cultured DCs（CD80/CD86,CD83/HLA-DR） were identified by flow cytometry.The ability of DCs to stimulate the proliferation of the allogeneic mixed lymphocyte was detected with methyl thiazolyl terazolium（MTT） assay.The IL-12 level in supernatant was determined by enzyme linked immunosorbent assay（ELISA ）.Results（1 ） Compared with Group C,observing with inverted microscope,the dendritic pseudopods arrounding cells of Group I was shorter and scarcer. （2） The expression of CD80/CD86 and CD83/HLA DR was（36.9±2.8）%and（12.9±2.4）%,（39.4±2.1 ）%and （14.4±3.0）%,（19.6±2.6）%and（7.1±1.4）%,respectively.Furthermore,there was statistical significance between I group and C group（P0.05）,while no statistical significance between M g oup and C group（P0.05）.（3）The capability to stimulate the mixed lymphocyte to proliferate was 1.44±0.23,1.37±0.10 and 0.56±0.12 respectively in group C, M and I,there was significant difference between group I and C（P0.05）,but no significant difference between group M and C（P0.05）.（4）The levels of IL-12 secreted by DCs was（958±321） pg/ml,（982±408）pg/ml and（377±312） pg/
出处
《疑难病杂志》
CAS
2011年第11期842-845,共4页
Chinese Journal of Difficult and Complicated Cases
基金
河北省高等学校强势特色学科(No.2005-52)
河北省卫生厅重点学科跟踪项目(No.GL200737)
关键词
七氟醚
树突状细胞
脐带血单核细胞
免疫功能
Sevoflurane
Dendritic cells
Cord blood mononuclear cytes
Immunity function
