循环白细胞ZDHHC24和FAM46C基因表达与冠心病发病风险的相关性

The correlation between ZDHHC24and FAM46Cgene expression levels and risk factors in patients with coronary artery disease

目的:芯片筛选结合在线数据库分析冠状动脉粥样硬化性心脏病(冠心病)共同差异表达基因,通过临床样本验证其表达变化差异,并揭示候选基因表达水平与冠心病发病风险的相关性。方法:数据筛选:病例组选取某医院心内科5名ST段抬高型心肌梗死(STEMI)患者,对照组选取同时期参与体检的3名志愿者,收集外周血,分离血浆外泌体,对其差异表达mRNA做高通量筛选;结合GEO数据集GSE62646、GSE59867分析共同差异表达基因。临床样本验证:病例组选取该医院心内科151例冠心病患者(含STEMI患者96例,稳定型冠心病患者55例)及85例健康者,qPCR检测外周血白细胞候选差异基因的表达情况;并对其在THP-1单核细胞泡沫化进程中的表达变化情况进行探索。结果:芯片分析结果显示:与健康对照组相比,STEMI患者血浆外泌体中ZDHHC24表达显著上调,FAM46C表达显著下调(均P<0.001);GEO数据集分析发现,与稳定型冠心病相比,STEMI患者外周血单个核细胞(PBMCs)中ZDHHC24表达显著上调,FAM46C表达显著下调(均P<0.05)。临床样本验证结果显示:与健康对照组及稳定型冠心病相比,STEMI患者ZDHHC24、FAM46C的表达水平均显著降低(均P<0.05);与健康对照组相比,稳定型冠心病患者ZDHHC24的表达水平显著降低(P<0.05)。多因素Logistic回归分析显示,ZDHHC24的低表达是独立于性别、年龄、血脂异常、高血压及糖尿病外稳定型冠心病与STEMI的发病风险因素(OR=0.272,95%CI=(0.166~0.445),P<0.001)。ROC曲线分析显示,ZDHHC24联合HDL-C和Apo-A1对冠心病具有较高的诊断特性(AUC分别为0.798,0.849;敏感度分别为60.3%,64.2%;特异度均为89.4%;均P<0.001)。此外,THP-1单核细胞巨噬化和泡沫化进程中ZDHHC24、FAM46CmRNA的表达水平显著上调(均P<0.05)。结论:外周血白细胞ZDHHC24、FAM46C的差异性表达可能参与冠心病的发展,ZDHHC24联合相关脂代谢指标可能对疾病的诊断提供参考价值。

Objective:Microarray screening combined with the online database to analyze common differentially expressed genes in coronary artery disease(CAD),verify the differences in expression changes through clinical samples,and reveal the correlation between the expression level of candidate genes and the risk of CAD.Method:Data screening:5 patients with ST-segment elevation myocardial infarction(STEMI) in a hosipital were selected in the case group,and 3 volunteers were selected in the control group to take part in the physical examination at the same time.Plasma exosomes were isolated,and high-throughput screening was performed on their differentially expressed mRNAs;combined with the GEO datasets GSE62646,GSE59867 to analyze common differentially expressed genes.Clinical sample verification:In this group,151 CAD patients(including 96 STEMI patients and 55 stable CAD patients) and 85 healthy patients were selected.The expression of candidate differential genes in peripheral blood leukocytes was detected by qPCR;and to explore the change of expression in the foaming process of THP-1 monocytes.Result:The results of microarray analysis showed that compared with the healthy control,the expression of ZDHHC24 in plasma exosomes was significantly upregulated,and the expression of FAM46 C was significantly downregulated(all P<0.001);GEO datasets analysis found that compared with stable CAD,STEMI patients the expression of ZDHHC24 in peripheral blood mononuclear cells(PBMCs)was significantly up-regulated,and the expression of FAM46 Cwas significantly downregulated(all P<0.05).The clinical sample verification results showed that compared with the healthy control group and stable CAD,the expression levels of ZDHHC24 and FAM46 Cin STEMI patients were significantly reduced(all P<0.05);compared with the healthy control group,the expression level of ZDHHC24 in stable CAD patients was significantly reduced(P<0.05).Multivariate logistic regression analysis showed that low expression of ZDHHC24 was a risk factor for CAD outside of gender,age,hyperlipidemia,hypertension,and diabetes(OR=0.272,95%CI=(0.166~0.445),P<0.001),The ROC analysis showed that ZDHHC24 combined with HDL-C,Apo-A1 had higher diagnostic characteristics for CAD(AUC were 0.798,0.849 respectively;sensitivity were 60.3%,64.2%respectively;specificity was 89.4%respectively;all P<0.001).In addition,ZDHHC24 and FAM46 CmRNA expression levels were significantly upregulated during THP-1 monocyte macrophage and foaming(all P<0.05).Conclusion:Differential expression of peripheral blood leukocytes ZDHHC24 and FAM46 Cmaybe involved in the development of CAD.ZDHHC24 combined with related lipid metabolism indicators may provide reference value for the diagnosis of diseases.