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目的检测血清中环状RNA hsa_circ_0029642的含量,明确其与肺动脉高压是否有密切关系,探讨循环环状RNA hsa_circ_0029642作为肺动脉高压血清标志物的可能性。方法收集2016年3-7月入院的先天性心脏病伴肺动脉高压患者血清20例(A组),先天性心脏病不伴肺动脉高压患者血清20例(B组),风湿性心脏病伴肺动脉高压患者血清20例(C组),各组从血清中提取总RNA并进行质量检测,进行反转录反应,获得相应的c DNA,以GAPDH作为内参进行标定,进行实时荧光定量PCR反应,根据各样本的定量PCR结果计算RQ值,绘制ROC曲线。结果 3组样本的总RNA均有较好的质量,A组与C组的RQ值差异无统计学意义(P>0.05),A组与B组的RQ值差异有统计学意义(P<0.05),B组与C组的RQ值差异有统计学意义(P<0.01)。即A组和C组的RQ值显著低于B组,而A组与C组的RQ值差异无统计学意义。ROC曲线(曲线下面积AUC=0.752,P<0.01,95%CI:0.624~0.884)。结论先天性心脏病肺动脉高压患者血清中环状RNA hsa_circ_0029642明显下调,循环环状RNA hsa_circ_0029642可作为肺动脉高压新的血清标志物。
Objective To detect the content of circulating RNA in human serum and determine whether it has a close relationship with pulmonary hypertension and to explore the possibility of circulating RNA as a serological marker of pulmonary hypertension. Methods Twenty patients (group A) with congenital heart disease and pulmonary arterial hypertension admitted from March to July 2016 were enrolled. Twenty patients (group B) with congenital heart disease without pulmonary hypertension were included in the study. Patients with rheumatic heart disease and pulmonary hypertension Twenty patients (group C) were selected. The total RNA was extracted from the serum and detected by mass spectrometry. Reverse transcription reaction was carried out to obtain the corresponding c DNA. GAPDH was used as an internal standard for real-time fluorescence quantitative PCR. RQ values were calculated using the quantitative PCR results of the samples and ROC curves were plotted. Results The total RNA of three groups had good quality. There was no significant difference in RQ between group A and group C (P> 0.05), and the difference of RQ between group A and group B was statistically significant (P <0.05 ), There was a significant difference in RQ between group B and group C (P <0.01). That is, the RQ values of group A and group C were significantly lower than those of group B, while the RQ values of group A and group C had no statistical significance. ROC curve (area under the curve AUC = 0.752, P <0.01, 95% CI: 0.624-0.884). Conclusions The serum level of hsa_circ_0029642 is significantly down-regulated in patients with congenital heart disease with pulmonary hypertension, and the circulating circular RNA hsa_circ_0029642 can be used as a new serum marker of pulmonary hypertension.