胆红素对脐血T淋巴细胞功能的影响

发布时间：2017-12-31 21:26

  本文关键词：胆红素对脐血T淋巴细胞功能的影响　出处：《青岛大学》2005年硕士论文　论文类型：学位论文

  更多相关文章： 胆红素 脐血 T淋巴细胞 CD25抗原 凋亡 γ-干扰素(IFN-γ) 白细胞介素-4(IL-4)

【摘要】：目的 探讨胆红素(Bilirubin)对脐血T淋巴细胞功能的影响。 方法 选择21例正常足月新生儿为研究对象,收集并分离脐血单个核细胞(CBMC),分别置于含有不同浓度胆红素(0mg/dl,6mg/dl,9mg/dl,12mg/dl)的牛血清白蛋白(BSA/PBS)溶液中进行预先孵育,洗涤后经植物血凝素(PHA)刺激培养48小时,采用丫锭橙/溴化乙锭(AB/EO)染色法测定细胞凋亡水平,间接免疫荧光法测定CBMC中CD25~+(IL-2受体)细胞百分率,酶联免疫吸附试验(ELISA)检测培养上清中IL-4及IFN-γ水平。 结果 ①胆红素浓度0mg/dl组、6mg/dl组、9mg/dl组、12mg/dl组细胞凋亡率分别为(28.72±11.09)%,(32.44±12.72)%,(56.36±9.24)%,(61.49±8.93)%。12mg/dl组显著高于6mg/dl组、Omg/dl组(q=12.55、14.16,p0.01),9mg/dl组亦显著高于6mg/dl组、0mg/dl组(q=10.34、11.94,p0.01),而12mg/dl组与9mg/dl组间比较细胞凋亡水平差异无显著性(q=2.22,p0.05),6mg/dl组与0mg/dl组比较细胞凋亡率亦无显著性差别(q=1.60,p0.05)。②胆红素浓度0mg/dl组、6mg/dl组、9mg/dl组、12mg/dl组CBMC体外经PHA刺激后CD25阳性细胞百分率分别为(64.25±6.18)%,(67.28±4.87)%,(22.12±4.61)%,(19.89±6.72)%。12mg/dl组CD25阳性细胞百分率明显低于6mg/dl组、0mg/dl组(q=38.35、35.90,p0.01),9mg/dl组亦明显低于6mg/dl组、0mg/dl组(q=36.54、34.09,p0.01),但12mg/dl组与9mg/dl组间比较无显著性差别(q=1.81,p0.05),且6mg/dl组与0mg/dl组比较CD25阳性细胞百分率亦无显著性差别(q=2.45,p0.05)。③CBMC体外经不同浓度胆红素作用后经PHA诱生IFN-γ水平12mg/dl组明显低于6mg/dl组、0mg/dl组(q=38.17、41.95,p0.01),9mg/dl组亦明显低于6mg/dl组、0mg/dl组(q=35.95、39.73,p0.01),6mg/dl组低于0mg/dl组,差异有统计学意义(q=3.78,p0.05),而12mg/dl组与9mg/dl组比较差异无显著性(q=2.21,p0.05)。④各组培养上清中IL-4产生水平比较均无显著性差异(p0.05)。⑤各组培养上清中IFN-γ/IL-4比值12mg/dl组明显低于6mg/dl组、0mg/dl组(q=13.28、16.23,p0.01),9mg/dl组明显低于6mg/dl组、0mg/dl组(q=12.49、15.44,p0.01),6mg/dl组低于0mg/dl组,差异有统计学意义(q=2.95,p0.05),但12mg/dl组与9mg/dl组比较差异无显著性(q=0.79,p0.05)。⑥经不同浓度胆红素作用的CBMC经48小时培养后,CD25阳性细胞百分率与凋亡细胞百分率呈显著负相关(r=-0.75,p0.01),培养上清中IFN-γ水平、IFN-γ/IL-4比值亦与凋亡细胞百分率呈显著负相关(r=-0.79、-0.69,p0.01),而培养上清中IL-4产生水平与细胞凋亡率无明显相关性(r=-0.20,
[Abstract]:Objective to investigate the effect of bilirubin (Bilirubin) on the function of T lymphocyte in umbilical cord blood.
Methods 21 cases of normal full-term newborns as the research object, collecting and separating umbilical cord blood mononuclear cells (CBMC), were placed with different concentrations of bilirubin (0mg/dl, 6mg/dl, 9mg/dl, 12mg/dl) of bovine serum albumin (BSA/PBS) solution were pre incubated after washing by phytohemagglutinin (PHA) stimulated for 48 hours. Ingot acridine orange / ethidium bromide (AB/EO) staining to determine apoptosis level, determination of CBMC CD25~+ in indirect immunofluorescence (IL-2 receptor) cells, enzyme-linked immunosorbent assay (ELISA) and IL-4 IFN- gamma detection levels in supernatant of culture.
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