The Expression of Serum MiR-29b，MiR-199a，MiR-19a-3p in Patients with Acute Cerebral Infarction And Their Relationship with Disease Severity And Prognosis
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摘要:
目的 探讨急性脑梗死（acute cerebral infarction，ACI）患者血清微小RNA-29b（miR-29b）、微小RNA-199a（miR-199a）、微小RNA-19a-3p（miR-19a-3p）表达及其与病情严重程度和预后的关系。 方法 （1）大鼠随机分为对照组（Control组）及模型组（Model组），每组6只，Model组大鼠采用改良Longa线栓法制作ACI模型，检测分析大鼠血清及脑组织中miR-29b、miR-199a、miR-19a-3p表达水平；（2）选取2023年6月至2024年6月在唐山市人民医院确诊的106例ACI患者（研究组），并选择同期在该院体检的108例健康人群为对照组。根据NIHSS评估结果，将研究组患者分为轻度组42例，中度组34例，重度组30例。按照mRS评估结果将患者分成预后良好组68例，预后不良组38例。实时荧光定量PCR法测定血清miR-29b、miR-199a、miR-19a-3p表达水平；通过Pearson相关性分析ACI患者血清miR-29b、miR-199a、miR-19a-3p水平与患者NIHSS评分、mRS评分的相关性；ACI患者预后不良的风险因素则采用多因素Logistic回归分析；ROC曲线分析血清miR-29b、miR-199a、miR-19a-3p水平及NHISS评分对ACI患者预后不良的预测价值。 结果 （1）与Control组比较，Model组大鼠血清及脑组织中miR-199a、miR-19a-3p水平明显上升，而miR-29b水平明显下降（P < 0.05）；（2）与对照组患者比较，伴随ACI患者病情逐渐加重，轻度组、中度组、重度组患者血清miR-199a、miR-19a-3p水平明显升高，miR-29b水平明显降低（P < 0.05）；预后不良组患者入院NHISS评分、miR-199a、miR-19a-3p水平显著高于预后良好组，miR-29b显著低于预后良好组（P < 0.05）；血清miR-199a、miR-19a-3p水平与NIHSS评分呈正相关；miR-29b水平与NIHSS评分呈负相关（P < 0.05）；血清miR-199a、miR-19a-3p水平升高，入院NHISS评分提高是ACI患者发生预后不良的独立危险因素，血清miR-29b水平升高是ACI患者发生预后不良的保护因素（P < 0.05）；血清miR-29b、miR-199a、miR-19a-3p水平及NHISS评分联合预测ACI患者预后不良的AUC为0.988；联合预测AUC优于单独预测（Z = 2.878，3.551，3.300，3.452，P < 0.05）。 结论 ACI患者血清miR-199a、miR-19a-3p表达显著上调，miR-29b显著下调，与患者病情及预后有关，三者联合预测患者预后不良具有一定价值。miR-29b、miR-199a、miR-19a-3p可作为评估ACI患者病情发展及预后的重要预测指标。 Abstract:Objective To investigate the expression of serum microRNA-29b （miR-29b）, microRNA-199a （miR-199a）, and microRNA-19a-3p （miR-19a-3p） in patients with acute cerebral infarction （ACI） and their relationship with disease severity and prognosis. Methods （1） Rats were randomly divided into a control group （Control group） and a model group （Model group）, with 6 rats in each group. ACI was induced in the model group using the modified Longa suture method; the expression levels of miR-29b, miR-199a, miR-19a-3p in rat serum and brain tissue were detected and analyzed. （2）A total of 106 ACI patients diagnosed at Tangshan People's Hospital from June 2023 to June 2024 were enrolled as the study group, and 108 healthy individuals who underwent physical check-ups at the same hospital during the same period were chosen as the control group. According to the NIHSS evaluation results, the study group patients were assigned into mild （42 cases）, moderate （34 cases）, and severe （30 cases） groups. Based on the mRS evaluation results, the patients were assigned into good prognosis （68 cases） and poor prognosis （38 cases） groups. Real-time fluorescence quantitative PCR was applied to measure the expression levels of serum miR-29b, miR-199a, and miR-19a-3p. Pearson correlation analyzed the correlation between serum miR-29b, miR-199a, miR-19a-3p levels and NIHSS score and mRS score in ACI patients. The risk factors for poor prognosis in ACI patients were analyzed using multivariate logistic regression. ROC curves were used to analyze the predictive value of serum miR-29b, miR-199a, miR-19a-3p levels and NHISS scores in poor prognosis of ACI patients. Results （1） Compared with the Control group, the levels of miR-199a and miR-19a-3p in the serum and brain tissue of the Model group rats were significantly increased, while the level of miR-29b was significantly decreased （P < 0.05）; （2） Compared with the control group, as the severity of ACI patients gradually increased, the levels of serum miR-199a and miR-19a-3p unusually increased in the mild, moderate, and severe groups, while the level of miR-29b was significantly decreased （P < 0.05）. The NHISS score, miR-199a and miR-19a-3p levels in the poor prognosis group were significantly higher than those in the good prognosis group, while miR-29b was significantly lower than that in the good prognosis group（P < 0.05）. The levels of serum miR-199a and miR-19a-3p were positively correlated with NIHSS score; the level of miR-29b was negatively correlated with NIHSS score （P < 0.05）. Elevated serum miR-199a and miR-19a-3p levels and increased NHISS score were independent risk factors for poor prognosis in ACI patients, and elevated serum miR-29b levels were protective factors for poor prognosis in ACI patients （P < 0.05）. The combined prediction of poor prognosis in ACI patients using serum miR-29b, miR-199a, miR-19a-3p levels and NHISS scores yielded an ACI of 0.988, which was superior to individual predictions （Z = 2.878, 3.551, 3.300, 3.452, P < 0.05）. Conclusion The serum expression levels of miR-199a and miR-19a-3p were significantly up-regulated, while miR-29b levels significantly down-regulated, which were related to the disease severity and prognosis. The the combination of the three miRNAs had certain value in predicting the poor prognosis of patients. MiR-29b, miR-199a, and miR-19a-3p can be used as important predictors to evaluate the progression and prognosis of ACI patients. -
Key words:
- Acute cerebral infarction /
- MicroRNA-29b /
- MicroRNA-199a /
- MicroRNA-20b /
- Disease severity /
- Prognosis
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图 1 血清miR-29b、miR-199a、miR-19a-3p水平及NHISS评分对ACI患者预后不良预测的ROC曲线
Figure 1. ROC curve of serum miR-29b，miR-199a，miR-19a-3p levels and NHISS score for predicting poor prognosis in ACI patients
表 1 引物序列
Table 1. Primer Sequence
基因 上游引物5'-3' 下游引物5'-3' miR-29b CTCAACTGGTGTCGTGGAGTCGGCAATTCAGTTGAG ACACTCCAGCTGGGTAGCACCATTTGAAATC miR-199a AAGGTGAAGGTCGGAGTCAAC GGGGTCATTGATGGCAACAATA miR-19a-3p GGCGGGGAAAGTGTGTGTCT GTGCAGTCGTGGCGTGTG U6 CATCCGTAAAGACCTCTATGCCAAC ATGGAGCCACCGATCCACA 
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表 2 大鼠血清及脑组织中miR-29b、miR-199a、miR-19a-3p水平比较（$\bar x \pm s $）
Table 2. Comparison of miR-29b，miR-199a，miR-19a-3p levels in rat serum and brain tissue （$\bar x \pm s $）
组别 n 血清 脑组织 miR-29b miR-199a miR-19a-3p miR-29b miR-199a miR-19a-3p Control组 6 1.02 ± 0.01 1.01 ± 0.01 1.01 ± 0.01 1.02 ± 0.02 1.02 ± 0.01 1.01 ± 0.01 Model组 6 0.72 ± 0.05 1.44 ± 0.11 1.56 ± 0.14 0.66 ± 0.04 1.38 ± 0.14 1.52 ± 0.13 t − 14.412 9.536 9.599 19.718 6.283 9.581 P − <0.001* <0.001* <0.001* <0.001* <0.001* <0.001* *P < 0.05。
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表 3 两组基线资料比较[（$\bar x \pm s $）/n（%）]
Table 3. Comparison of baseline data between two groups [（$\bar x \pm s $）/n（%）]
组别 n 年龄 性别（男） BMI（kg/m2） 研究组 106 64.37 ± 9.45 54（50.94） 26.05 ± 2.72 对照组 108 65.96 ± 9.21 59（54.63） 25.87 ± 2.64 χ2/t − 1.246 0.292 0.491 P − 0.214 0.589 0.624
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表 4 各组患者血清miR-29b、miR-199a、miR-19a-3p水平比较（$\bar x \pm s $）
Table 4. Comparison of serum miR-29b，miR-199a，and miR-19a-3p levels among different patient groups （$\bar x \pm s $）
组别 n miR-29b miR-199a miR-19a-3p 对照组 108 1.01 ± 0.11 1.04 ± 0.12 1.21 ± 0.15 轻度组 42 0.73 ± 0.08a 1.47 ± 0.15a 2.01 ± 0.21a 中度组 34 0.64 ± 0.07b 1.56 ± 0.16b 2.19 ± 0.23b 重度组 30 0.57 ± 0.06bc 1.72 ± 0.18bc 2.36 ± 0.25bc F − 275.366 265.441 460.552 P − <0.001* <0.001* <0.001* *P < 0.05；与对照组比较，aP < 0.05；与轻度组比较，bP < 0.05；与中度组比较，cP < 0.05；与重度组比较，dP < 0.05。
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表 5 血清miR-29b、miR-199a、miR-19a-3p水平与NIHSS评分、mRS评分的相关性分析
Table 5. Correlation analysis of serum miR-29b，miR-199a，miR-19a-3p levels with NIHSS score and mRS score
项目 miR-29b miR-199a miR-19a-3p r P r P r P NIHSS评分 −0.495 <0.001* 0.598 <0.001* 0.612 <0.001* mRS评分 −0.526 <0.001* 0.605 <0.001* 0.557 <0.001* *P < 0.05。
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表 6 ACI患者预后不良的单因素分析[（$\bar x \pm s $）/n（%）]
Table 6. Univariate analysis of poor prognosis in ACI patients [（$\bar x \pm s $）/n（%）]
一般资料 预后不良组（n=38） 预后良好组（n=68） χ2/t P 性别 2.177 0.140 男 23（60.53） 31（45.59） 女 15（39.47） 37（54.41） 年龄（岁） 65.15 ± 9.17 63.93 ± 8.59 0.684 0.495 BMI（kg/m2） 26.22 ± 2.78 25.97 ± 2.69 0.453 0.651 发病至溶栓时间（min） 164.63 ± 17.02 159.57 ± 16.03 1.524 0.130 脑梗死部位 0.586 0.900 脑干 7（18.42） 16（23.53） 基底节区 18（47.37） 29（42.65） 脑叶 7（18.42） 14（20.59） 多部位并存 6（15.79） 9（13.24） 糖尿病 0.021 0.885 有 19（50.00） 35（51.47） 无 19（50.00） 33（48.53） 高血压 1.477 0.224 是 22（57.89） 31（45.59） 否 16（42.11） 37（54.41） LDL-C（mmol/L） 1.71 ± 0.18 1.67 ± 0.17 1.137 0.258 HDL-C（mmol/L） 1.29 ± 0.13 1.34 ± 0.16 1.646 0.103 TG（mmol/L） 1.73 ± 0.19 1.67 ± 0.18 1.613 0.110 TC（mmol/L） 5.83 ± 0.61 5.64 ± 0.58 1.588 0.115 FBG（mmol/L） 6.75 ± 0.68 6.69 ± 0.67 0.440 0.661 WBC（×109/L） 8.45 ± 0.87 8.23 ± 0.84 1.277 0.205 CRP（mmol/L） 113.45 ± 11.48 109.81 ± 11.02 1.607 0.111 Hb（g/L） 128.71 ± 12.93 130.65 ± 13.09 0.735 0.464 ALB（g/L） 39.76 ± 4.02 40.87 ± 4.09 1.348 0.181 Scr（mmol/L） 90.12 ± 9.04 88.73 ± 8.91 0.766 0.445 UA（μmol/L） 301.27 ± 30.18 292.64 ± 29.37 1.437 0.154 入院NHISS评分（分） 9.03 ± 0.98 7.89 ± 0.81 6.438 <0.001* mRS评分（分） 4.26 ± 0.74 1.66 ± 0.23 26.831 <0.001* miR-29b 0.58 ± 0.07 0.67 ± 0.07 6.348 <0.001* miR-199a 1.71 ± 0.18 1.49 ± 0.16 6.489 <0.001* miR-19a-3p 2.36 ± 0.25 2.06 ± 0.22 6.409 <0.001* *P < 0.05。
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表 7 ACI患者预后不良影响因素的多因素Logistic回归分析
Table 7. Multivariate Logistic regression analysis of factors affecting poor prognosis in ACI patients
影响因素 B SE Waldχ2 OR 95%CI P miR-29b −0.350 0.092 14.436 0.705 0.589～0.844 <0.001* miR-199a 0.943 0.267 12.477 2.568 1.522～4.334 <0.001* miR-19a-3p 0.889 0.256 12.052 2.432 1.472～4.017 <0.001* 入院NHISS评分 0.720 0.213 11.420 2.054 1.353～3.118 <0.001* 常量 −12.350 2.764 20.177 0.000 − <0.001* *P < 0.05。
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表 8 血清miR-29b、miR-199a、miR-19a-3p水平及NHISS评分对ACI患者预后不良预测的ROC结果
Table 8. ROC results for predicting poor prognosis in ACI patients based on serum miR-29b，miR-199a，miR-19a-3p levels and NHISS score
指标 敏感度（%） 特异性（%） 截断值 AUC 95%CI 约登指数 miR-29b 90.11 78.47 0.62 0.894 0.819～0.945 0.686 miR-199a 81.58 75.00 1.57 0.857 0.776～0.917 0.566 miR-19a-3p 91.31 72.59 2.2 0.886 0.809～0.939 0.627 NHISS评分 89.47 79.41 8.41 0.896 0.821～0.947 0.689 联合 95.27 99.96 0.86 0.988 0.945～0.985 0.999
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