基于外泌体多miRNA表达水平构建的子痫前期风险预测模型效能分析

doi:10.11958/20230485

摘要/Abstract

摘要：

目的探讨基于多个外泌体微小RNA（miRNA）表达水平构建子痫前期（PE）风险模型的可行性，并验证其对PE的预测效能。方法选择2019年6月—2021年12月建档且孕周≤20周的孕妇1 037例作为研究对象，入组后收集所有患者首次建档的一般资料及实验室检查资料。通过qRT-PCR分析所有样本中外泌体miRNA表达情况（包括miR-155-5p、miR-215-5p、miR-203a-3p、miR-199a-5p及miR-125a-3p）。而后对所有患者随访至妊娠结束，统计随访期间PE发生情况并根据结果将患者分为PE组及对照组。使用Cox回归分析PE的影响因素，以ggrisk包构建多miRNA风险模型，并采用受试者工作特征（ROC）曲线评估所构建模型对PE的预测效果。结果截至2022年10月31日随访结束，最终完成随访974例（93.92%）。PE组65例和对照组909例。PE组年龄、孕前体质量指数（BMI）、孕12周腰围均大于对照组，吸烟史及饮酒史占比高于对照组（P<0.05）。PE组三酰甘油、低密度脂蛋白胆固醇、总胆固醇、丙氨酸转氨酶、天冬氨酸转氨酶、血小板分布宽度、平均血小板体积、血红蛋白、miR-155-5p、miR-199a-5p、miR-215-5p水平均高于对照组，而促甲状腺激素、miR-125a-3p及miR-203a-3p水平低于对照组（P<0.05）。miR-125a-3p、miR-155-5p、miR-199a-5p、miR-203a-3p及miR-215-5p表达水平为PE发生的独立预测因子（P<0.05）。由上述外泌体miRNA构建的风险预测模型在PE发生方面具有良好的预测价值（AUC=0.998，P<0.001），敏感度为96.92%（63/65），特异度为93.94%（854/909）。结论 miR-125a-3p、miR-155-5p、miR-199a-5p、miR-203a-3p及miR-215-5p与PE的发生相关，以上述外泌体中5个miRNA构建的PE预测模型效果较好。

关键词: 先兆子痫, 外泌体, 微RNAs, 预测模型

Abstract:

Objective To investigate the feasibility of constructing a preeclampsia (PE) risk model based on multiple exosomal micrornas (miRNA) expression levels and to verify its efficacy in predicting PE. Methods A total of 1 037 pregnant women who were archived in our hospital from June 2019 to December 2021 and whose gestational weeks were less than or equal to 20 weeks were selected as the research subjects. The expression of exosomal miRNA (including miR-155-5p, miR-215-5p, miR-203a-3p, miR-199a-5p and miR-125a-3p) in all samples was detected by qRT-PCR. Then, all patients were followed up to the end of pregnancy. The occurrence of PE during the follow-up period was counted, and all samples were divided into the PE group and the control group according to results. Cox regression was used to analyze the influencing factors of PE. The multi-miRNA risk model was constructed with ggrisk package, and the predictive effect of the model on PE was evaluated by receiver operating characteristic (ROC) curve. Results By the end of follow-up on October 31, 2022, 974 cases were finally followed up, and the follow-up completion rate was 93.92%. Among all the 974 patients who completed the follow-up, 65 patients developed PE, so they were finally divided into the PE group, and 909 cases were used as the control group. The age, pre-pregnancy BMI and waist circumference at 12 weeks of gestation were higher in the PE group than those in the control group (P<0.05). The proportions of smoking history and drinking history were higher in the PE group than those of the control group (P<0.05). The contents of triglyceride (TG), low density lipoprotein cholesterol (LDL-C), total cholesterol (TC), alanyl aminotransferase (ALT), aspartate aminotransferase (AST), platelet distribution width (PDW), mean platelet volume (MPV), miR-155-5p, miR-199a-5p and miR-215-5p were higher in the PE group than those in the control group，while contents of thyroid stimulating hormone (TSH), miR-125a-3p and miR-203a-3p were lower in the PE group than those in the control group (P<0.05). The expression levels of miR-125a-3p, miR-155-5p, miR-199a-5p and miR-215-5p were independent predictors of PE (P<0.05). The predictive risk model constructed from the above miRNAs had good predictive value in the occurrence of PE (AUC=0.998), with a sensitivity of 98.46% (63/65) and a specificity of 93.94% (854/909). Conclusion miR-125a-3p, miR-155-5p, miR-199a-5p, miR-203a-3p and miR-215-5p are significantly related to the occurrence of PE, and the PE prediction model constructed with the above five miRNAs has better effect.

Key words: pre-eclampsia, exosomes, microRNAs, predict model

中图分类号:

R714.252

图/表 7

参考文献 24

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基因名称	引物序列（5′→3′）	产物大小/bp
miR-125a- 3p	上游：ACGTGTGTAGCTTATCAGACTG	22
miR-125a- 3p	下游：AATGGTTGTTCTCCACACTCTC	22
miR-155- 5p	上游：GAGGGTTAATGCTAATCGTGATAGG	25
miR-155- 5p	下游：GCACAGAATCAACACGACTCACTAT	25
miR-199a- 5p	上游：TTATTACCCAGGCAGACACCG	21
miR-199a- 5p	下游：AGTGCGAACTGTGGCGATGC	21
miR-203a- 3p	上游：CCGGTGAAATGTTTAGGACC	20
miR-203a- 3p	下游：GCCGCGTGAAAATGTTTAGG	20
miR-215- 5p	上游：CTCGAGATGTCATCCTCAG	19
miR-215- 5p	下游：GAATTCGTGAGTTCTTCTG	19
U6	上游：CTTCGGCAGCACATATAC	18
U6	下游：GAACGCTTCACGAATTTG	18

基因名称	引物序列（5′→3′）	产物大小/bp
miR-125a- 3p	上游：ACGTGTGTAGCTTATCAGACTG	22
miR-125a- 3p	下游：AATGGTTGTTCTCCACACTCTC	22
miR-155- 5p	上游：GAGGGTTAATGCTAATCGTGATAGG	25
miR-155- 5p	下游：GCACAGAATCAACACGACTCACTAT	25
miR-199a- 5p	上游：TTATTACCCAGGCAGACACCG	21
miR-199a- 5p	下游：AGTGCGAACTGTGGCGATGC	21
miR-203a- 3p	上游：CCGGTGAAATGTTTAGGACC	20
miR-203a- 3p	下游：GCCGCGTGAAAATGTTTAGG	20
miR-215- 5p	上游：CTCGAGATGTCATCCTCAG	19
miR-215- 5p	下游：GAATTCGTGAGTTCTTCTG	19
U6	上游：CTTCGGCAGCACATATAC	18
U6	下游：GAACGCTTCACGAATTTG	18

组别	n	年龄/岁	孕前BMI/ （kg/m²）	孕12周腰围/cm	饮酒史	吸烟史	高血压	收缩压/mmHg	舒张压/mmHg	孕≥2次
对照组	909	27.28±3.89	21.53±1.62	77.54±3.83	67（7.4）	90（9.9）	114（12.5）	115.66±6.68	77.51±5.42	288（31.7）
PE组	65	29.85±6.46	21.99±3.07	80.97±8.67	10（15.4）	14（21.5）	9（13.8）	115.31±8.33	76.89±4.34	26（40.0）
t或χ²		4.895^＊＊	2.027^＊	6.189^＊＊	5.351^＊	8.631^＊＊	0.093	0.408	0.902	1.921

组别	n	年龄/岁	孕前BMI/ （kg/m²）	孕12周腰围/cm	饮酒史	吸烟史	高血压	收缩压/mmHg	舒张压/mmHg	孕≥2次
对照组	909	27.28±3.89	21.53±1.62	77.54±3.83	67（7.4）	90（9.9）	114（12.5）	115.66±6.68	77.51±5.42	288（31.7）
PE组	65	29.85±6.46	21.99±3.07	80.97±8.67	10（15.4）	14（21.5）	9（13.8）	115.31±8.33	76.89±4.34	26（40.0）
t或χ²		4.895^＊＊	2.027^＊	6.189^＊＊	5.351^＊	8.631^＊＊	0.093	0.408	0.902	1.921

组别		n	FPG/（mmol/L）		TG/（mmol/L）		LDL-C/（mmol/L）			TC/（mmol/L）		ALT/（U/L）		AST/（U/L）
对照组		909	5.11±1.41		1.89±0.47		3.08±0.60			5.64±0.89		23.24±5.42		18.41±4.11
PE组		65	5.22±1.98		2.05±0.72		3.31±1.15			5.91±1.89		25.16±8.42		19.75±4.31
t			0.584		2.558^＊		2.756^＊＊			2.111^＊		2.643^＊＊		2.524^＊
组别	PLT/（×10⁹/L）			Hb/（g/L）		PDW/%		MPV/fL	TSH/（U/L）		FT3/（pmol/L）		FT4/（pmol/L）
对照组	214.59±37.01			123.43±9.33		15.30±2.04		8.77±2.01	2.25±0.39		4.25±0.42		12.56±1.91
PE组	219.36±57.39			126.47±10.35		17.60±4.27		9.98±2.96	1.95±0.34		4.29±0.64		12.95±3.21
t	0.961			2.519^＊		7.933^＊＊		4.516^＊＊	6.070^＊＊		0.664		1.529