摘要
目的观察脐血单核细胞(CBMC)经不同水平胆红素孵育后髓样分化蛋白2(MD-2)、TNF受体相关因子6(TRAF6)mRNA表达的变化。方法采集10例健康足月新生儿的脐血,经纤维蛋白结合法分离得CBMC,将其加入含不同水平胆红素(0μmol.L-1、17.1μmol.L-1、42.8μmol.L-1、102.6μmol.L-1、153.9μmol.L-1、220.6μmol.L-1和307.8μmol.L-1)的牛血清清蛋白溶液中孵育1 h,再加入脂多糖(LPS)刺激30 min。收集CBMC,采用反转录-PCR方法测定其MD-2、TRAF6 mRNA的表达水平。结果 1.LPS单独作用对CMBC MD-2 mRNA和TRAF6 mRNA表达水平无明显影响,低水平胆红素(17.1μmol.L-1、42.8μmol.L-1)单独作用对CBMC MD-2 mRNA和TRAF6 mRNA表达水平无明显影响。2.先经不同水平胆红素孵育,再接受LPS刺激,胆红素有抑制细胞MD-2 mRNA表达的趋势,随着胆红素水平的升高,抑制趋势越明显;先经不同水平胆红素孵育,再接受LPS刺激,低水平胆红素促进细胞TRAF6 mRNA的表达,当胆红素≥102.6μmol.L-1时,抑制TRAF6 mRNA表达,胆红素水平越高,抑制作用越明显。在相同胆红素水平作用下,随着MD-2 mRNA表达水平的降低,TRAF6 mRNA表达水平随之降低。结论低水平胆红素对CBMC MD-2 mRNA的表达无明显影响,对TRAF6 mRNA的表达有促进作用。高水平胆红素对二者的表达均有抑制作用,随着胆红素水平升高,抑制作用越来越明显。高胆红素血症患儿免疫功能低下可能与高浓度胆红素对免疫细胞的抑制作用有关。
Objective To observe the changes of myeloid differentiation protein 2（MD-2） and tumor necrosis factor receptor-associa-ted factor 6（TRAF6） mRNA expressions after the cord blood monocytes（CBMC） were incubated with different concentrations of bilirubin. Methods Cord blood of 10 cases of healthy full-term newborns were collected,the CBMC separated by the method of gelatin/plasma coated flasks were preincubated with various concentrations of bilirubin（0 μmol·L-1,17.1 μmol·L-1,42.8 μmol·L-1,102.6 μmol·L-1,153.9 μmol·L-1,220.6 μmol·L-1 and 307.8 μmol·L-1） dissolved in bovine albumin solution for 1 hour,then were further stimulated with lipopolysaccharide（LPS） for 30 min.After collecting CBMC,the expressions of MD-2 and TRAF6 in CBMC were measured by reverse transcription-PCR. Results 1.LPS had no significant effect on mRNA expression levels of MD-2 and TRAF6 of CBMC alone,and also low concentrations of bilirubin（17.1 μmol·L-1,42.8 μmol·L-1） had no significant effect on mRNA expression levels of MD-2 and TRAF6 of CBMC.2.At first,CBMC were incubated with different concentrations of bilirubin,and then accepted the stimulation of LPS,the trend of MD-2 mRNA expression of CBMC inhibited by bilirubin.The higher the concentration,the more obvious the trends.CBMC were incubated with different concentrations of bilirubin firstly,and then accepted the stimulation of LPS,low concentrations of bilirubin promoted TRAF6 mRNA expression of CBMC;when the bilirubin concentration ≥ 102.6 μmol·L-1,TRAF6 mRNA expression of CBMC were inhibited.The higher the concentration of bilirubin,the more obvious the inhibition.Under the same concentration of bilirubin,the changes of the MD-2 mRNA expression levels of CBMC were declined trend as TRAF6 mRNA expression levels decreased. Conclusions The low concentration of bilirubin don′t significantly affect on the MD-2 mRNA expression and promotes the TRAF6 mRNA expression.High concentration of bilirubin inhibits the expression of both,with the bilirubin concentration increased,the more obvious inhibition.Low immune function in newborns with hyperbilirubinemia may be related to high concentrations of the inhibitory effect of bilirubin on the immune cells.
出处
《实用儿科临床杂志》
CAS
CSCD
北大核心
2012年第2期91-94,共4页
Journal of Applied Clinical Pediatrics
基金
四川省科技厅科研基金资助项目(05JY029-054-2
07SG004-005)
关键词
胆红素
脐血
单核细胞
髓样分化蛋白2
肿瘤坏死因子受体相关因子6
bilirubin
cord blood
monocyte
myeloid differentiation protein 2
tumor necrosis factor receptor-associated factor 6
