miR-151a-3p在急性脑梗死患者血清中的表达及与炎性因子的相关性研究

The expression of serum miR-151a-3p in patients with acute cerebral infarction and its correlation with pro-inflammatory factors

目的：探讨急性脑梗死（ACI）患者外周血微小RNA（miRNA）表达的临床意义及与炎性因子的相关性，为其在ACI诊断和治疗方面提供新的依据及思路。方法采用回顾性研究方法，选择2004年4月至7月武汉大学人民医院神经内科收治的114例首次发病且病程为2～14d的ACI患者临床资料，以同期年龄、性别相匹配的58例健康体检者为健康组。记录ACI患者的脑梗死危险因素，以及所有研究对象的血清miR-151a-3p、白细胞介素（IL-6、IL-8）、C-反应蛋白（CRP）、肿瘤坏死因子-α（TNF-α）水平。分析血清miR-151a-3p水平与脑梗死面积和分型、梗死原因及炎性因子的关系；并分析不同miR-151a-3p表达水平ACI患者的10年存活率。结果114例ACI患者中男性59例，女性55例；年龄48～63岁，平均（55.0±6.7）岁；大梗死25例，中梗死26例，小梗死53例，腔隙性梗死10例。按改良急性卒中治疗低分子肝素试验（TOAST）分型，动脉粥样硬化性血栓形成性脑梗死（AT）92例，心源性脑栓塞（CE）10例，小动脉闭塞性脑梗死（SAD）12例。排除脑梗死危险因素对miRNA表达水平的影响后，ACI组血清miR-151a-3p表达水平明显高于健康组（2-ΔΔCt：2.28±1.85比1.27±0.98，P＜0.01）；大梗死组、中梗死组、小梗死组和腔隙性梗死组血清miR-151a-3p表达水平均高于健康组（2-ΔΔCt：1.78±1.02、1.92±1.11、2.22±1.54、2.61±1.82比1.27±0.98，均P＜0.05），不同梗死面积组间比较差异无统计学意义；AT组、CE组血清miR-151a-3p表达水平较健康组明显升高（2-ΔΔCt：2.01±1.45、1.99±0.89比1.27±0.98，均P＜0.05），而SAD组与健康组比较差异无统计学意义（2-ΔΔCt：1.72±0.30比1.27±0.98，P＞0.05）。ACI组血清IL-6、IL-8、CRP和TNF-α水平均明显高于健康组〔IL-6（ng/L）：45.21±14.33比39.70±13.15，IL-8（μg/L）：29.12±14.92比22.50±10.12，CRP（mg/L）：6.61±3.02比5.40±2.75，TNF-α（ng/L）：65.20±16.14比55.70±14.35，均P＜0.05〕。相关性分析显示，ACI患者血清miR-151a-3p与IL-6、IL-8、CRP、TNF-α均呈显著正相关（R2值分别为0.092、0.055、0.034、0.036，均P＜0.05）。miR-151a-3p低表达组患者10年存活率较miR-151a-3p高表达组明显升高〔以1.27±0.98为界值，48.57%（17/35）比34.18%（27/79），log-rank＝3.411，P＝0.045〕。结论血清miR-151a-3p水平可提示脑梗死的发病及进展，有可能作为诊断ACI的潜在生物学标志物。

Objective To investigate the clinical significance of serum microRNA-151a-3p （miR-151a-3p） expression in peripheral blood of patients with acute cerebral infarction （ACI）, and to analyze the correlation between miR-151a-3p and related inflammatory factors, in order to obtain new evidence and ideas in the diagnosis and treatment of ACI. Methods A retrospective analysis was conducted. The clinical data of patients with ACI admitted to Department of Neurology of People＆#39;s Hospital of Wuhan University from April to July in 2004 were enrolled. 114 ACI patients with first onset and duration of 2-14 days served as the research objects, and in the same period 58 healthy persons with matched age, and gender served as healthy control group. The risk factors of cerebral infarction in ACI patients and levels of serum miR-151a-3p, interleukins （IL-6, IL-8）, C-reactive protein （CRP）, tumor necrosis factor-α （TNF-α） in all the subjects were completely recorded. The correlation between serum miR-151a-3p and the area and type of cerebral infarction, the causes of infarction as well as the inflammatory cytokines was analyzed. The correlation of 10-year survival rate of patients with different expression levels of miR-151a-3p in patients with ACI was analyzed. Results A total of 114 patients with ACI were enrolled, with 59 male, 55 female, and age ranged 48-63 years with a mean of （55.0±6.7） years. Large infarction was found in 25 cases, middle sized infarction in 26 cases, small infarction in 53 cases, and lacunar infarction in 10 cases. According to the modified Trial of Org 10172 in acute stroke treatment （TOAST）, the patients were classified as thrombotic cerebral infarction （AT） 92 cases, embolism （CE） from cardiac origin 10 cases, and small arterial occlusive cerebral infarction （SAD） 12 cases. After eliminating the influence of cerebral infarction risk factors on the expression level of miRNAs, and compared with that of healthy control group, the level of serum miR-151a-3p expression was significantly increased in ACI group （2-ΔΔCt： 2.28±1.85 vs. 1.27±0.98, P 〈 0.01）; the levels of serum miR-151a-3p in large, middle, small, lacunar infarction groups were markedly up-regulated （2-ΔΔCt： 1.78±1.02, 1.92±1.11, 2.22±1.54, 2.61±1.82 vs. 1.27±0.98, all P 〈 0.05） with no significant difference among different infarction groups. The serum miR-151a-3p expression in AT and CE groups was significantly higher than that of the healthy control group （2-ΔΔCt： 2.01±1.45, 1.99±0.89 vs. 1.27±0.98, both P 〈 0.05）, but no significant difference was found between SAD group and healthy control group （2-ΔΔCt： 1.72±0.30 vs. 1.27±0.98, P 〉 0.05）. The levels of serum IL-6, IL-8, CRP and TNF-α in ACI group were all higher than those of healthy control group [IL-6 （ng/L）： 45.21±14.33 vs. 39.70±13.15, IL-8 （μg/L）： 29.12±14.92 vs. 22.50±10.12, CRP （mg/L）： 6.61±3.02 vs. 5.40±2.75, TNF-α （ng/L）： 65.20±16.14 vs. 55.70±14.35, all P 〈 0.05]. In addition, higher expression of serum pro-inflammatory mediators IL-6, IL-8, CRP and TNF-α were positively correlated with miR-151a-3p （R2 value were 0.092, 0.055, 0.034, 0.036, all P 〈 0.05）. Ten-year survival rate was higher in patients with low expression of miR-151a-3p [with 1.27±1.98 as the boundary, 48.57% （17/35） vs. 34.18% （27/79）, log-rank = 3.411, P = 0.045]. Conclusions Up-regulated serum miR-151a-3p may be involved in the pathophysiology of ACI. Therefore, miR-151a-3p may be used as a reference to predict the severity of neurological deficit in clinic.