阻塞性睡眠呼吸暂停低通气综合征患者血清miRNA-92a水平与血管内皮功能损伤的关系研究

Study on the serum miRNA-92a level and vascular endothelial function injury in OSAHS patients

目的明确阻塞性睡眠呼吸暂停低通气综合征(OSAHS)部分通过微小RNA(mi RNA)-92a介导损伤血管内皮细胞。方法收集2018年1月至12月接受睡眠呼吸检测的患者,测定其血清中mi RNA-92a水平,分析其与OSAHS相关指标的关系。同时体外培养内皮细胞,OSAHS血清刺激观察内皮细胞形态变化及JC-1染色结果,进一步明确内皮细胞损伤情况;采用逆转录聚合酶链反应(RT-PCR)和蛋白免疫印迹(Western blot)方法检测mi RNA-92a靶基因变化以进一步明确内皮细胞损伤机制。结果接受睡眠呼吸检测的患者72例,其中非OSAHS组22例,轻度OSAHS组18例,中度OSAHS组10例,重度OSAHS组22例。OSAHS患者血清mi RNA-92a水平显著升高,且随OSAHS严重程度加重mi RNA-92a水平亦升高。体外实验中OSAHS血清可显著损伤内皮细胞,内皮细胞肿胀排列紊乱,边界不清楚。JC-1染色显示绿色荧光较对照组显著增强。RT-PCR及Western blot检测提示重度OSAHS血清刺激后的内皮细胞Krüppel样因子-2(KLF-2)、Krüppel样因子-4(KLF-4)及内皮型一氧化氮合酶(e NOS)表达显著降低。结论OSAHS患者血清mi RNA-92a显著增加,作用于其靶基因KLF-2、KLF-4及e NOS,减少靶基因表达,影响内皮细胞线粒体功能,损伤内皮细胞。

Objective To clarify that the vascular endothelial cell injury caused by obstructive sleep apnoea hypopnea syndrome(OSAHS)is partly mediated by mi RNA-92 a.Methods Serum mi RNA-92 a level was measured in patients who underwent polysomnography between January 2018 and December 2018.The correlation between mi RNA-92 a and OSAHS was analyzed.Meanwhile,endothelial cells were cultured in vitro,and morphological changes and JC-1 staining results of endothelial cells were observed after OSAHS serum stimulation,so as to further clarify the injury of endothelial cells.The changes of mi RNA-92 a target gene were detected by reverse transcription-polymerase chain reaction(RT-PCR)and Western blot to further clarify the mechanism of endothelial cell injury.Results Seventy-two patients received polysomnography,including 22 cases in the non-OSAHS group,18 in the mild OSAHS group,10 in the moderate OSAHS group,and 22 in the severe OSAHS group.Serum mi RNA-92 a level was significantly increased in the OSAHS patients,and it also increased with the aggravation of OSAHS severity.OSAHS serum significantly damaged endothelial cells.Endothelial cells were swollen,disordered arrangement,and unclear boundaries.JC-1 staining showed that green fluorescence was significantly enhanced compared with the control group.RT-PCR and Western blot showed that the expressions of Krüppel-like factor-2(KLF-2),Krüppel-like factor-4(KLF-4)and endothelial nitric oxide synthase(e NOS)were significantly decreased under OSAHS serum stimulation.Conclusion Serum mi RNA-92 a of OSAHS patients is significantly increased,and reduces the expression of target genes KLF-2,KLF-4 and e NOS,affects the mitochondrial function of endothelial cells,and injures endothelial cells.