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[脐血单个核细胞中Toll样受体的表达及意义。]

[Expression and significance of Toll-like receptors in cord blood mononuclear cells.].

作者信息

Wang Lin, Xu Jian-bo, Tian Yuan, Liu Ya-lan, Wu He-shui

机构信息

Department of Pediatrics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China.

出版信息

Zhonghua Er Ke Za Zhi. 2007 May;45(5):365-8.

Abstract

OBJECTIVE

To investigate the expression of TLR4/2 mRNA in neonatal cord blood mononuclear cells (MNC).

METHODS

Forty-six neonates without asphyxia and 40 neonates with asphyxia were divided into groups depending on the gestational age. In the neonates without asphyxia, there were 18 full term infants (the gestational age > or = 37 weeks), 16 preterm infants whose gestational age was > or = 32 weeks but < 37 weeks, and 12 preterm infants whose gestational age was < 32 weeks. In the neonates with asphyxia, 11 were full term infants, 15 were preterm infants whose gestational age was > or = 32 weeks but < 37 weeks and 14 were preterm infants at gestational age < 32 weeks. MNCs were separated and cultured with LPS (1 microg/ml) for 3 h. Cells were collected for analysis of gene expression of TLR4/2 by RT-PCR technique. Cell supernatants were taken to measure TNF-alpha production following the ELISA protocol. Fifteen healthy adults were enrolled into the control group. In addition, the Pearson correlation analyses were carried out between the levels of TLR4, TLR2 mRNA and the levels of TNF-alpha.

RESULTS

In the neonates without asphyxia, TLR4, TLR2 mRNA and TNF-alpha levels were 0.75 +/- 0.12, 0.63 +/- 0.08, 2502.6 +/- 273.1 ng/L, separately, in the full term infants, 0.37 +/- 0.04, 0.32 +/- 0.03, 1218.8 +/- 145.7 ng/L, separately, in the preterm infants whose gestational ages were > or = 32 weeks but < 37 weeks, and 0.26 +/- 0.03, 0.20 +/- 0.03, 811.8 +/- 105.2 ng/L separately, in the preterm infants whose gestational ages were < 32 weeks. In the neonates with asphyxia, TLR4, TLR2 mRNA and TNF-alpha levels were 0.58 +/- 0.07, 0.50 +/- 0.06, 1946.4 +/- 244.2 ng/L, separately, in the full term infants, 0.29 +/- 0.03, 0.26 +/- 0.03, 970.0 +/- 94.3 ng/L, separately, in the preterm infants whose gestational age was > or = 32 weeks but < 37 weeks, and 0.17 +/- 0.02, 0.14 +/- 0.02, 652.6 +/- 60.3 ng/L, separately, in the preterm infants whose gestational age was < 32 weeks. The levels of TLR4, TLR2 mRNA and TNF-alpha in the adults were 2.71 +/- 0.75, 2.61 +/- 0.33, 9270.1 +/- 1098.3 ng/L, separately. In the preterm infants and full term infants, the levels of TLR4, TLR2 mRNA and TNF-alpha were lower in comparison to the adults. The lower the gestational age, the lower the levels of TLR4, TLR2 mRNA and TNF-alpha. There were significant differences between the levels of TLR4, TLR2 mRNA and TNF-alpha of the neonates without asphyxia and those of the neonates with asphyxia. In the neonates with asphyxia, the levels of TLR4, TLR2 mRNA and TNF-alpha were lower than those in the neonates without asphyxia (P < 0.01). Whether the neonates were asphyxic or not, the levels of TLR4, TLR2 were paralleled with the levels of TNF-alpha.

CONCLUSIONS

The expression of TLRs in the neonates, especially in the preterm infants was lower than that in the adults, which probably contributes to the susceptibility of neonates to infections.

摘要

目的

研究新生儿脐血单个核细胞(MNC)中TLR4/2 mRNA的表达。

方法

46例无窒息新生儿和40例窒息新生儿按胎龄分组。无窒息新生儿中,18例足月儿(胎龄≥37周),16例胎龄≥32周但<37周的早产儿,12例胎龄<32周的早产儿。窒息新生儿中,11例足月儿,15例胎龄≥32周但<37周的早产儿,14例胎龄<32周的早产儿。分离MNC并用脂多糖(1μg/ml)培养3小时。收集细胞采用RT-PCR技术分析TLR4/2基因表达。按照ELISA方案取细胞上清液检测TNF-α的产生。15名健康成年人纳入对照组。此外,对TLR4、TLR2 mRNA水平与TNF-α水平进行Pearson相关分析。

结果

无窒息新生儿中,足月儿TLR4、TLR2 mRNA和TNF-α水平分别为0.75±0.12、0.63±0.08、2502.6±273.1 ng/L;胎龄≥32周但<37周的早产儿分别为0.37±0.04、0.32±0.03、1218.8±145.7 ng/L;胎龄<32周的早产儿分别为0.26±0.03、0.20±0.03、811.8±105.2 ng/L。窒息新生儿中,足月儿TLR4、TLR2 mRNA和TNF-α水平分别为0.58±0.07、0.50±0.06、1946.4±244.2 ng/L;胎龄≥32周但<37周的早产儿分别为0.29±0.03、0.26±0.03、970.0±94.3 ng/L;胎龄<32周的早产儿分别为0.17±0.02、0.14±0.02、652.6±60.3 ng/L。成年人TLR4、TLR2 mRNA和TNF-α水平分别为2.71±0.75、2.61±0.33、9270.1±1098.3 ng/L。早产儿和足月儿中,TLR4、TLR2 mRNA和TNF-α水平低于成年人。胎龄越低,TLR4、TLR2 mRNA和TNF-α水平越低。无窒息新生儿与窒息新生儿的TLR4、TLR2 mRNA和TNF-α水平存在显著差异。窒息新生儿中,TLR4、TLR2 mRNA和TNF-α水平低于无窒息新生儿(P<0.01)。无论新生儿是否窒息,TLR4、TLR2水平与TNF-α水平平行。

结论

新生儿尤其是早产儿中TLRs表达低于成年人,这可能是新生儿易感染的原因。

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