MicroRNA-15b与糖尿病视网膜病变患者外周血VEGF及相关炎症因子的关系

Relationship between micro ribonucleic acid-15b and vascular endothelial growth factor and related inflammatory factors in peripheral blood of diabetic retinopathy patients

目的分析microRNA-15b(miR-15b)与糖尿病视网膜病变(DR)患者外周血血管内皮生长因子(VEGF)及相关炎症因子的关系。方法前瞻性研究。选取2019年6月至2021年3月就诊于本院的140例糖尿病患者,按DR临床分期标准分组,即无DR(NDR)组、单纯型DR(SDR)组、增生型DR(PDR)组,另选30名同期本院健康体检者作为健康对照(NC)组。采用RT-PCR法测定血清miR-15b表达,通过ELISA法测定血清VEGF及炎症因子[细胞间黏附分子-1(ICAM-1)、白细胞介素(IL)-1β、IL-8、高敏C反应蛋白(hs-CRP)、肿瘤坏死因子-α(TNF-α)]水平,采用流式细胞仪测定DR患者外周血微血管内皮功能指标[内皮祖细胞(EPC)、循环祖细胞(CPC)、内皮细胞(CEC)]比例。采用Pearson相关分析DR患者血清miR-15b与VEGF、炎症反应指标、微血管内皮功能指标的相关性;绘制受试者工作特征曲线(ROC),分析miR-15b对DR的预测价值。结果4组受检者miR-15b、VEGF、炎症因子及微血管内皮功能指标水平相比,差异均有统计学意义(均为P<0.05),其中血清miR-15b表达、EPC、CPC水平由低至高分别为PDR组、SDR组、NDR组、NC组。血清VEGF、ICAM-1、IL-1β、IL-8、hs-CRP、TNF-α及CEC水平由高至低分别为PDR组、SDR组、NDR组、NC组;血清miR-15b表达与VEGF、ICAM-1、IL-1β、IL-8、hs-CRP、TNF-α、CEC均呈负相关(均为P<0.05),与EPC、CPC均呈正相关(均为P<0.05)。miR-15b预测DR发生的AUC为0.869(95%CI为0.796~0.924),诊断临界值为0.68,灵敏度、特异度分别为82.15%、79.50%,约登指数为0.617。结论DR患者血清miR-15b表达降低,与外周血VEGF及炎症因子水平密切相关,且在微血管损伤及炎症反应中均有重要参与作用。

Objective To analyze the relationship between micro ribonucleic acid-15b(miR-15b)and vascular endothelial growth factor(VEGF)and related inflammatory factors in the peripheral blood of patients with diabetic retinopathy(DR).Methods A prospective study was conducted.Totally 140 patients with diabetes mellitus who visited our hospital from June 2019 to March 2021 were grouped according to the clinical staging criteria of DR into the non-DR(NDR)group,simple DR(SDR)group and proliferative DR(PDR)group.In addition,30 healthy subjects who underwent physical examination in our hospital during the same period were selected as the negative control(NC)group.The expression of miR-15b in the serum was determined by real-time fluorescent quantitative PCR,and the levels of VEGF and inflammatory factors[intercellular adhesion molecule-1(ICAM-1),interleukin-1β(IL-1β),interleukin-8(IL-8),high-sensitivity C-reactive protein(hs-CRP)and tumor necrosis factor-α(TNF-α)]in the serum were determined by enzyme-linked immunosorbent assay.The proportion of microvascular endothelial function indexes[endothelial progenitor cells(EPC),circulating progenitor cells(CPC),and circulating endothelial cells(CEC)]in the peripheral blood was determined by a flow cytometer.Pearson correlation analysis was used to determine the correlation between miR-15b and VEGF,inflammatory response indexes as well as microvascular endothelial function indexes.The receiver operating characteristic curve was drawn to analyze the predictive value of miR-15b to DR.Results The levels of miR-15b,VEGF,inflammatory indexes and microvascular endothelial function indexes among the four groups showed statistically significant differences(all P<0.05).The expression levels of miR-15b,EPC and CPC in the serum showed an increasing trend in the PDR group,SDR group,NDR group and NC group.The levels of VEGF,ICAM-1,IL-1β,IL-8,hs-CRP,TNF-αand CEC in the serum showed a decreasing trend in the PDR group,SDR group,NDR group and NC group.The expression level of miR-15b was negatively correlated with VEGF,ICAM-1,IL-1β,IL-8,hs-CRP,TNF-α,and CEC(all P<0.05),and positively correlated with EPC and CPC(both P<0.05).The area under the curve for MiR-15b in predicting the DR was 0.869(95%CI was 0.796-0.924),the diagnosis threshold was 0.68,sensitivity and specificity were 82.15%and 79.50%,respectively,and the Youden index was 0.617.Conclusion The decrease of serum mir-15b expression in DR patients is closely related to the levels of VEGF and inflammatory factors in peripheral blood and plays an essential role in microvascular injury and inflammatory response.