新生儿缺氧缺血性脑病患儿外周血单核细胞Toll样受体3和微小RNA-210的表达水平及意义

Expression levels and significance of TLR3 and miRNA-210 in peripheral blood mononuclear cells of neonates with hypoxic-ischemic encephalopathy

目的探讨新生儿缺氧缺血性脑病(HIE)患儿外周血单核细胞Toll样受体3(TLR3)和微小RNA-210(miR-210)表达水平及临床意义,试图为临床早期诊断及治疗新生儿HIE提供新的靶点。方法选取2015年3月-2017年3月四川绵阳四○四医院收治的67例HIE新生儿为研究对象,其中重度HIE 21例,中度HIE 26例,轻度HIE 20例,同期选取25例四川绵阳四○四医院出生的足月健康新生儿为对照组,应用实时荧光定量PCR(qRT-PCR)检测受试新生儿外周血单核细胞中TLR3、miR-210表达水平。结果新生儿出生后1 d,HIE重度、中度、轻度组TLR3、miR-210 mRNA水平均明显高于对照组,差异有统计学意义(均P<0.05),且随着疾病严重程度增加,TLR3、miR-210 mRNA水平呈明显上升趋势,各疾病等级之间两两比较,差异有统计学意义(均P<0.05),新生儿出生后3 d、7 d,HIE组TLR3、miR-210 mRNA水平呈明显下降趋势,差异有统计学意义(均P<0.05);HIE重度、中度、轻度组新生儿行为神经(NBNA)评分明显低于对照组,差异有统计学意义(均P<0.05),且随着疾病严重程度增加,NBNA评分呈明显下降趋势,差异有统计学意义(P<0.05);HIE患儿外周血单核细胞中TLR3、miR-210水平与NBNA评分呈明显负性相关(r=-0.396、-0.457,均P<0.05),HIE患儿外周血单核细胞中TLR3与miR-210表达呈明显正相关(r=0.493,P<0.05)。结论 TLR3、miR-210在HIE新生儿外周血单核细胞中呈明显高表达,二者有望成为HIE早期诊断、临床分度及判断预后的参考指标。

Objective To explore the expression levels and significance of Toll-like receptor 3（ TLR3） and miRNA-210 in peripheral blood mononuclear cells of neonates with hypoxic-ischemic encephalopathy（ HIE）,provide a new target for clinical early diagnosis and treatment of neonatal HIE. Methods Sixty-seven HIE neonates treated in Sichuan Mianyang 404 Hospital from March 2015 to March 2017 were selected as study object,including 21 neonates with severe HIE,26 neonates with moderate HIE,and 20 neonates with mild HIE. Twenty-five healthy full-term neonates born in Sichuan Mianyang 404 Hospital during the same period were selected as control group. Real-time fluorescence quantitative PCR was used to detect the expression levels of TLR3 and miRNA-210 in peripheral blood mononuclear cells. Results At one day after birth,the levels of TLR3 and miR-210 mRNA in severe HIE group,moderate HIE group,and mild HIE group were statistically significantly higher than those in control group（ P〈0. 05）. With the increase of severity of the disease,the levels of TLR3 and miR-210 mRNA showed a significantly increasing trend,there were statistically significant differences among the neonates with different pathological grades（ P〈0. 05）. At 3 and 7 days after birth,the levels of TLR3 and miR-210 mRNA in neonates with HIE decreased significantly,there were statistically significant differences（ P〈0. 05）. NBNA scores in severe HIE group,moderate HIE group,and mild HIE group were statistically significantly lower than that in control group（ P〈0. 05）. With the increase of severity of the disease,NBNA scores showed a significantly decreasing trend,there were statistically significant differences among the neonates with different pathological grades（ P〈0. 05）. The levels of TLR3 and miR-210 in peripheral blood mononuclear cells of neonates with HIE were significantly negatively correlated with NBNA score（ r =-0. 396,P〈0. 05; r =-0. 457,P〈0. 05）. TLR3 was significantly positively correlated with miR-210 expression in peripheral blood mononuclear cells of neonates with HIE（ r = 0. 493,P〈0. 05）. Conclusion The expression levels of TLR3 and miR-210 in peripheral blood mononuclear cells of neonates with HIE are high; TLR3 and miR-210 are expected to be reference indexes for early diagnosis,clinical grading,and prognosis evaluation of HIE.