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目的研究粘蛋白域蛋白-3(TiM-3)在先天性心脏病患儿外周血中的表达水平。方法临床纳入广东省深圳市南山区妇幼保健院2014年3月至2015年3月期间儿科收治的62例先天性心脏病患儿为研究组,另同期选取62例正常健康儿童为对照组。采用多色流式技术和荧光实时定量PCR(q RT-PCR)技术检测两组研究对象外周血单核细胞亚群表面上的TiM-3分子以及TiM-3 m RNA表达水平。采用脂多糖(LPS)与重组人galectin-9刺激两组对象外周血单核细胞,利用酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)测定血清中IL-1β,IL-6以及TNF-α水平,探讨TiM-3与先天性心脏病的关系。结果观察组单核细胞TiM-3,CD14+CD16+TiM-3,CD14+CD16-TiM-3以及单核细胞TiM-3 m RNA表达水平均明显高于对照组,两组之间差异有统计学意义(t=22.635,8.407,24.447,7.087,均P<0.05)。LPS+galectin-9组IL-1β,IL-6以及TNF-α水平均明显高于LPS组和空白组,三组间差异有统计学意义(F=853.30,102.84,506.45,P<0.05)。结论先天性心脏病患儿TiM-3表达水平明显上升,可能诱导单核细胞炎症的分泌从而引起疾病的发生与发展。
Objective To study the expression of mucin domain protein-3 (TiM-3) in peripheral blood of children with congenital heart disease. Methods Sixty-two children with congenital heart disease admitted to Pediatrics from March 2014 to March 2015 in Nanshan District MCH Hospital of Shenzhen City, Guangdong Province were enrolled in the study. Another 62 healthy children were selected as the control group. The expression of TiM-3 mRNA and TiM-3 mRNA on the surface of peripheral blood mononuclear cell subgroups were detected by multi-color flow cytometry and real-time quantitative PCR (q RT-PCR) Serum levels of IL-1β, IL-6 and TNF-α were measured by enzyme linked immunosorbent assay (ELISA) Level, to explore the relationship between TiM-3 and congenital heart disease. Results The expression levels of TiM-3, CD14 + CD16 + TiM-3, CD14 + CD16-TiM-3 and monocyte TiM-3 mRNA in monocytes were significantly higher than those in the control group Significance (t = 22.635,8.407,24.447,7.087, all P <0.05). The levels of IL-1β, IL-6 and TNF-α in LPS + galectin-9 group were significantly higher than those in LPS group and blank group. There was significant difference among the three groups (F = 853.30,102.84,506.45, P <0.05). Conclusion The expression of TiM-3 in children with congenital heart disease obviously increases, which may induce the secretion of monocyte inflammation and cause the occurrence and development of the disease.