论文部分内容阅读
目的 :探讨测定急性呼吸窘迫综合征早产儿血浆中mir-124-3p和SPOCK2蛋白的临床意义,为早产儿严重肺损伤寻找新的高危因子和治疗靶标。方法:根据早产儿呼吸窘迫综合征(respiratory distress syndrome,RDS)诊断标准,早产儿分成RDS组17例及对照组(无RDS等并发症的普通早产儿)21例。采用ELISA和Western blot法测定外周血浆中和体外培养的人肺成纤维细胞中的SPOCK2蛋白。采用Target Scan分析与SPOCK2基因3′末端非编码区结合的可能mi RNA,并用荧光实时定量RTPCR扩增候选mi RNA。采用Pearson统计方法对血浆中SPOCK2蛋白和mi RNA进行相关分析。构建mir-124-3p慢病毒载体,并转染人肺成纤维细胞,研究mir-124-3p过表达对SPOCK2蛋白表达的影响。结果:对照组早产儿血浆中SPOCK2蛋白浓度为(7.24±0.43)μg/L,而RDS组升高为(16.43±0.54)μg/L,两组间比较差异有统计学意义(t=12.81,P<0.01)。SPOCK2基因3′末端存在mir-124-3p、mir-25-3p、mir-122-5p等候选的mi RNA结合位点。荧光定量RT-PCR证实外周血中不能测到mir-25-3p、mir-122-5p。mir-124-3p表达在对照组中较高,在RDS组中显著降低,差异有统计学意义(P<0.05)。并且mir-124-3p的表达量与SPOCK2蛋白血浆水平呈负相关(r=-0.645 9,P=0.012 6)。mir-124-3p慢病毒载体转染可以显著抑制人肺成纤维细胞中SPOCK2蛋白的表达。结论:测定早产儿血浆中SPOCK2蛋白,可能对RDS等肺疾病的病情评估有一定临床意义。利用mir-124-3p调控SPOCK2蛋白表达,可能对早产儿RDS等严重肺损伤的治疗有潜在应用价值。
Objective: To investigate the clinical significance of detecting mir-124-3p and SPOCK2 proteins in the plasma of premature infants with acute respiratory distress syndrome and to find new high-risk factors and therapeutic targets for severe lung injury in premature infants. Methods: According to the diagnostic criteria of respiratory distress syndrome (RDS) in preterm infants, premature infants were divided into RDS group (n = 17) and control group (n = 21). The SPOCK2 protein in human lung fibroblasts cultured in vitro and in vitro was determined by ELISA and Western blot. Target Scan was used to analyze possible mi RNAs that bind to the non-coding region of the 3 ’end of the SPOCK2 gene and candidate mi RNAs were amplified using real-time fluorescence quantitative RT-PCR. Pearson statistical method was used to analyze the correlation between SPOCK2 protein and miRNA in plasma. The mir-124-3p lentiviral vector was constructed and transfected into human lung fibroblasts to study the effect of mir-124-3p overexpression on SPOCK2 protein expression. Results: The SPOCK2 protein concentration in the control group was (7.24 ± 0.43) μg / L and that in RDS group was (16.43 ± 0.54) μg / L, the difference was statistically significant (t = 12.81, P <0.01). The miRNA binding sites of mir-124-3p, mir-25-3p and mir-122-5p were selected at the 3’-end of SPOCK2 gene. Fluorescent quantitative RT-PCR confirmed that mir-25-3p and mir-122-5p could not be detected in peripheral blood. The expression of mir-124-3p was higher in the control group and significantly lower in the RDS group, the difference was statistically significant (P <0.05). And the expression of mir-124-3p was negatively correlated with SPOCK2 protein plasma level (r = -0.6459, P = 0.012 6). Transfection with mir-124-3p lentiviral vector significantly inhibited SPOCK2 protein expression in human lung fibroblasts. Conclusion: The determination of SPOCK2 protein in plasma of preterm infants may have certain clinical significance in the evaluation of the disease such as RDS and other lung diseases. The use of mir-124-3p to regulate SPOCK2 protein expression may have potential value in the treatment of severe lung injury such as RDS in premature infants.