文章摘要
TLR9在不同胎龄新生儿脐血中的表达变化
TLR9 expression in umbilical cord blood of newborns at different gestational age
投稿时间：2016-01-18  
DOI：10.3969/j.issn.1000-0399.2016.10.004
中文关键词: Toll样受体9  新生儿  早产  脐血  胎龄
英文关键词: Toll-like receptors 9(TLR9)  Neonates  Preterm birth  Umbilical cord blood  Gestational age
基金项目:上海市嘉定区卫计委资助项目（项目编号：KYXM2010-12-9）
作者单位E-mail
王晓莉 201821 上海市嘉定区妇幼保健院新生儿科  
刘燕 201821 上海市嘉定区妇幼保健院新生儿科  
何振娟 200092 上海交通大学医学院附属新华医院新生儿科  
陈超 201102 复旦大学附属儿科医院新生儿科 chen6010@163.com 
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中文摘要:
      目的 通过观察早产儿不同胎龄Toll样受体9（TLR9）的表达，探讨早产儿免疫功能低下的机制。方法 采集2010年7月至2014年6月在上海市嘉定区妇幼保健院产科出生的活产新生儿的脐血229份，按胎龄分为4组，28~31周组，31~34周组，34~37周组，≥37周组，采用流式细胞术和实时荧光定量PCR方法，分别检测其TLR9的蛋白和mRNA表达情况，了解其与胎龄之间的关系，并分析mRNA和蛋白表达间的相关性。结果 TLR9阳性细胞率在28~31周组，31~34周组，34~37周组，≥37周组分别为（15.93±6.23）%，（11.63±6.70）%，（13.66±6.88）%，（20.51±12.06）%；其在胎龄28~31周较高，至31~34周逐渐下降至最低，两组差异有统计学意义（P<0.05）；34~37周后TLR9阳性细胞率表达逐渐升高，至≥37周达最高，两胎龄组比较，差异具有统计学意义（P<0.05）。31~37周间新生儿脐血TLR9阳性细胞率与胎龄呈正相关（r=0.273，P=0.006）。TLR9 mRNA表达在28~31周组，31~34周组，34~37周组，≥37周组分别为（4.95±3.44）%，（8.89±8.49）%，（13.91±10.92）%，（7.19±7.11）%；其在28~36周逐渐升高，与胎龄呈正相关（r=0.355，P< 0.001）。≥37周TLR9 mRNA表达量下降，该值虽高于28~31周，但差异无统计学意义（P>0.05）。相关性分析表明，同胎龄时期同样本新生儿的TLR9 mRNA和TLR9阳性细胞率之间存在负相关（r=-0.227，P=0.011）。结论 TLR9阳性细胞率和TLR9 mRNA表达在不同胎龄组新生儿间有差异，TLR9阳性细胞率表达在31~37周间随着胎龄的增加而增加，TLR9 mRNA在28~36周间随着胎龄的增加而增加。
英文摘要:
      Objective To investigate the mechanism of the weakened immune system in newborns via observing the effect of TLR9 on newborns at different gestational age. Methods We collected umbilical cord blood from 229 neonates at different gestational age from July 2010 to June 2014. According to the gestational age, the samples were divided into 4 groups, which were 28-31 weeks group, 31-34 weeks group, 34-37 weeks group and ≥37 weeks group. Flow cytometry was used to detect TLR9 positive cells of cord blood mononuclear(CBMC) and real-time quantitative PCR was applied to determine mRNA expression of TLR9 in order to analyze the relationship between those with gestational age. In the mean time, the correlation between mRNA and protein expressions of TLR9 was analysed. Results The rate of TLR9 positive cell was 15.93±6.23%(28-31 weeks), 11.63±6.70%(31-34 weeks), 13.66±6.88%(34-37 weeks), 20.51±12.06%(≥37 weeks), respectively. The rate of TLR9 positive cells was significantly higher in 28-31W group than in 31-34W group(P<0.05). TLR9 positive cell rate increased gradually and reached the highest in 37W group with significantly statistical difference(P<0.05). There was a positive correlation between the rate of TLR9 positive cell and gestational age from 31 weeks to 37 weeks, (r=0.273, P=0.006). The mRNA expression of TLR9 was 4.95±3.44%(28-31 weeks), 8.89±8.49%(31-34 weeks), 13.91±10.92%(34-37 weeks), 7.19±7.11%(≥37 weeks), respectively. The mRNA expression increased with gestational age from 28 weeks to 36 weeks,and it had positive correlation with gestational age(r=0.355, P<0.001). The mRNA expression of TLR9 decreased in ≥37 weeks group, and there was no significant difference compared with 28-31 weeks group(P>0.05). There was a negative correlation between the mRNA expression of TLR9 and the rate of TLR9 positive cell at the same gestational age in preterm infants(r=-0.227, P=0.011).Conclusion The rate of TLR9 positive cell and the mRNA expression of TLR9 are significantly different in newborns at different gestational age. With gestational age increasing, the rate of TLR9 positive cell rises from 31weeks to 37weeks, while the mRNA expression TLR9 is augmented from 28 weeks to 36 weeks.
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