瑞舒伐他汀对急性冠脉综合征患者CD4^+T淋巴细胞miRNA表达的影响

Differences of CD4^+ T lymphocyte miRNA gene expression in acute coronary artery syndrome(ACS) patients and the effects of rosuvastatin on its expressions

目的研究瑞舒伐他汀对急性冠脉综合征(ACS)患者外周血CD4+T淋巴细胞基因中微小RNA(miRNA)表达谱的影响,探讨瑞舒伐他汀钙治疗ACS的机制。方法选取2012年3-7月入住济南军区总医院的9例ACS患者,另以因疑似不典型冠心病症状而行320CT示冠脉病变≤50%的住院患者9例作为正常对照组。利用密度梯度法离心分离ACS患者及对照者的外周血单核细胞,免疫磁珠法进一步分离出CD4+T淋巴细胞。采用miRNA基因芯片技术检测CD4+T淋巴细胞miRNA的表达谱,筛选ACS患者与正常对照组,以及ACS患者用药前后差异表达的miRNA。选取其中差异性最为显著的3个miRNA,采用实时荧光定量聚合酶链反应(RT-PCR)对差异表达的miRNA进行验证。结果通过基因芯片共检测基因1900余条,其中有差异的有300余条。与正常对照比较,ACS患者中显著上调的基因有126条,如miRNA-21、miRNA-142、miRNA-20a等,显著下调的基因有202条,如miRNA-4734、miRNA-1182、miRNA1273f等。ACS患者服用瑞舒伐他汀钙(20mg,1次/d)10d后,与用药前相比,显著上调的基因有157条,如miRNA-4734、miRNA-1182、miRNA-663b等,显著下调的基因有137条,如miRNA-4789、miRNA-5692c、miRNA-26a等。采用RT-PCR进一步验证,结果与miRNA芯片检测结果一致。结论瑞舒伐他汀钙可能通过调节miRNA-4734、miRNA-1182、miRNA-4789等miRNA的表达从而发挥其对ACS的治疗作用。

Objective To investigate the effects of rosuvastatin on the expression profile of peripheral blood CD4~ T Iymphocytes miRNA gene in the patients with acute coronary syndrome （ACS） and screen the differentially expressed miRNAs before and after treatment, and elucidate the mechanisms of rosuvastatin calcium in the treatment of patients with ACS. Methods Nine cases were selected from the patients with ACS treated in the General Hospital of Jinan Military Command from Mar. to Jul. of 2012, with other 9 cases selected as controls, whose degree of coronary artery stenosis was less than 50% as confirmed by 320CT. Peripheral blood mononuclear cells were isolated by density gradient centrifugation, and CD4~ T lymphocytes were isolated by immunomagnetic beads method, miRNAs of CD4＋ T lymphocytes were detected by miRNA gene chip technology. The differentially expressed miRNAs between ACS patients and normal control, and those in ACS patients before and after treatment were screened. Three of the maximum difference miRNAs were selected and followed by verification by real-time polymerase chain reaction （RT-PCR）. Results More than 1900 genes were detected by gene microarray, of which more than 300 genes showed significant differences in expression. Comparing the ACS patients and normal controls, 126 genes were significantly up-regulated, including miRNA-21, miRNA-142, and miRNA-20a; and 202 genes were significantly down-regulated, including miRNA-4734, miRNA-1182, and miRNA-1273f. A total of 157 genes were significantly up-regulated after treatment with rosuvastatin calcium （20mg/d x 10d）,such as miRNA-4734, miRNA-1182, and miRNA-663b; and 137 genes were significantly down-regulated, such as miRNA-4789, miRNA-5692c, and miRNA-26a. The results were validated by RT-PCR and they were same as miRNA microarray. Conclusion Rosuvastain may play a role in the treatment of patients with ACS by regulating a series of miRNAs such as miRNA-4734, miRNA- 1182, and miRNA-4789.