不稳定型心绞痛气虚血瘀证患者外泌体MicroRNA差异表达

Differential Expression of MicroRNA in the Exosomes of Unstable Angina Patients with Qi Deficiency and Blood Stasis Syndrome

目的分析冠心病不稳定型心绞痛(UA)气虚血瘀证患者血清外泌体中MicroRNA(miRNA)的差异表达。方法收集115例UA患者(气虚血瘀证患者59例、非气虚血瘀证患者56例)和36名健康志愿者的外周血液,提取上清中的外泌体。采用高通量测序方法,检测9例气虚血瘀证、9例非气虚血瘀证UA患者以及6名健康志愿者血清外泌体中miRNA,分析他们之间差异表达的miRNA检测结果,确定目标miRNA。再采用qRT-PCR检测方法对未作高通量测试的97例UA患者(50例气虚血瘀证、47例非气虚血瘀证)及30名健康志愿者进行验证。结果(1)经过高通量测序后分析发现,与健康志愿者比较,miR-125b(P=0.032)、miR-10b(P=0.038)在UA患者血清外泌体中呈高表达。与UA非气虚血瘀证患者比较,miR-125b(P=0.028)、miR-10b(P=0.043)在气虚血瘀证患者血清外泌体中呈高表达。确定miR-125b、miR-10b为本研究的血清外泌体中目标miRNA。(2)对目标miRNA进行qRT-PCR检测验证发现,UA患者血清外泌体中miR-125b、miR-10b表达量高于健康志愿者,UA气虚血瘀证患者血清外泌体中miR-125b、miR-10b表达量高于非气虚血瘀证患者。使用ROC曲线下面积(AUC)进一步分析,提示miR-125b、miR-10b的ROC AUC分别为0.807、0.818,表明miR-125b、miR-10b高表达与UA气虚血瘀证之间存在一定关联。结论UA气虚血瘀证与非气虚血瘀证患者血清外泌体中存在差异表达的miRNA,其中,miR-125b、miR-10b在UA气虚血瘀证患者血清外泌体中呈高表达。

Objective To analyze differential expression of MicroRNA(miRNA)in serum exosomes of unstable angina(UA)patients.Methods Peripheral blood of 115 UA patients[including 59 patients with qi deficiency and blood stasis syndrome(QDBSS)and 56 patients without QDBSS]and 36 healthy volunteers were recruited,and the exosomes in the supernatant were extracted.The miRNA expression in the serum exosomes of 9 UA patients with QDBSS,9 patients without QDBSS,and 6 healthy volunteers were selected to perform high-throughput sequencing.Differentially expressed miRNA among the three groups to determine the target miRNA.According to the target miRNA,97 UA patients(50 patients with QDBSS and 47 patients without QDBSS)and 30 healthy volunteers without high-throughput sequencing were verified by qRT-PCR.Results(1)It was found from high-throughput sequencing that miR-125b(P=0.032)and miR-10b(P=0.038)were highly expressed in serum exosomes of UA patients than those of healthy volunteers,which were higher in UA patients with QDBSS than those without QDBSS(miR-125b,P=0.028;miR-10b,P=0.043).Therefore,miR-125b and miR-10b were determined as the target miRNA in the serum exosomes.(2)qRT-PCR showed,miR-125b and miR-10b were validated to be highly expressed in serum exosomes of UA patients than those of healthy volunteers,which were higher in UA patients with QDBSS than those without QDBSS.Further analysis showed that area under curve(AUC)of ROC of miR-125b and miR-10b were 0.807 and 0.818 respectively,which indicated that there was a certain correlation between the high expression of miR-125b and miR-10b and QDBSS of UA patients.Conclusions There were the differentially expressed miRNA in the serum exosomes of the UA patients with/without QDBSS.In particular,miR-125b and miR-10b showed high expression in the serum exosomes of the UA patients with QDBSS.