目的 检测微小RNA-138(miR-138)、微小RNA-338(miR-338)在痉挛型脑性瘫痪患儿中表达水平,并探究两者的临床价值。方法 选取2018年3月—2020年4月于本院门诊就诊及住院治疗的64例痉挛型脑性瘫痪患儿为研究对象(痉挛型脑性瘫痪组);同期选取62例健康体检儿童作为健康对照组。采用实时荧光定量PCR(qRT-PCR)法检测各组受试儿童血清中miR-138、miR-338表达水平;采用酶联免疫吸附(ELISA)法检测各组受试儿童血清肿瘤坏死因子-α(TNF-α)、C-反应蛋白(CRP)、神经生长因子(NGF)、内皮素(ET)水平;采用Pearson法分析痉挛型脑性瘫痪患儿血清miR-138、miR-338与TNF-α、CRP、NGF、ET表达水平相关性;采用受试者工作特征曲线(ROC)分析血清miR-138、miR-338水平对痉挛型脑性瘫痪的诊断价值;采用多因素Logistic回归分析影响痉挛型脑性瘫痪发生的因素。结果 痉挛型脑性瘫痪组血清中miR-138、miR-338、TNF-α、CRP、ET表达水平高于健康对照组,NGF表达水平低于健康对照组,差异均有统计学意义(t=9.598、9.592、11.039、42.397、14.172、5.744,P＜0.05)。痉挛型脑性瘫痪患儿血清miR-138与miR-338呈正相关(r=0.687,P＜0.05),且二者均与TNF-α、CRP、ET水平呈正相关(miR-138:r=0.519、0.483、0.494;miR-138:r=0.506、0.492、0.516,P＜0.05),与NGF水平呈负相关(r=-0.522、-0.496,P＜0.05)。血清miR-138、miR-338诊断痉挛型脑性瘫痪的曲线下面积(AUC)分别为0.882、0.858,截断值分别为1.189、1.406,特异性分别为75.8%、93.5%,敏感度分别为87.5%、67.2%;二者联合诊断痉挛型脑性瘫痪的AUC为0.950,特异性为90.3%,敏感度为93.8%。Logistic回归显示miR-138、miR-338是痉挛型脑性瘫痪发生的危险因素[OR(95%CI)分别为2.280(1.742~2.983)、2.222(1.637~3.016)],NGF是痉挛型脑性瘫痪发生的保护因子[OR(95%CI)为0.720(0.527~0.984),P＜0.05]。结论 痉挛型脑性瘫痪患儿血清中miR-138、miR-338表达水平显著上调,二者对痉挛型脑性瘫痪的发生、发展可能有重要提示作用。
Abstract
Objective To detect the expression levels of microRNA-138 (miR-138) and microRNA-338 (miR-338) in children with spastic cerebral palsy and explore the clinical value. Methods A total of 64 children with spastic cerebral palsy who were treated in the Third Affiliated Hospital of Zhengzhou University from March 2018 to April 2020 were selected into spastic cerebral palsy group. Meanwhile,62 healthy children were selected as healthy control group. The expression levels of miR-138 and miR-338 were detected by real-time quantitative PCR (qRT-PCR),the serum levels of tumor necrosis factor-α (TNF-α),C-reactive protein (CRP),nerve growth factor (NGF) and endothelin (ET) were detected by enzyme-linked immunosorbent assay (ELISA). Pearson method was used to analyze the correlation between serum miR-138,miR-338 and TNF-α,CRP,NGF,ET expression levels in children with spastic cerebral palsy Receiver operating characteristic curve (ROC) was used to analyze the diagnostic value of serum miR-138 and miR-338 in spastic cerebral palsy. Multivariate Logistic regression analysis was used to analyze the influencing factors of spastic cerebral palsy. Results The expression levels of miR-138,miR-338,TNF-α,CRP and ET in serum in spastic cerebral palsy group were higher than those in healthy control group,while NGF expression level was lower than that in control group(t=9.598,9.592,11.039,42.397,14.172,5.744,P＜0.05). There was a positive correlation between serum miR-138 and miR-338 in children with spastic cerebral palsy (r = 0.687,P < 0.05),and they were positively correlated with TNF-α,CRP,ET levels (miR-138:r=0.519,0.483,0.494;miR-338:r=0.506,0.492,0.516,P＜0.05),and negatively correlated with NGF level (r=-0.522,-0.496,P＜0.05). The area under the curve (AUC) of serum miR-138 and miR-338 in the diagnosis of spastic cerebral palsy was 0.882 and 0.858,the cut-off value was 1.189 and 1.406,the specificity was 75.8%,93.5%,and the sensitivity was 87.5% and 67.2%,respectively.The AUC of the combined diagnosis was 0.950,the specificity was 90.3%,and the sensitivity was 93.8%. Logistic regression analysis that miR-138,miR-338 and NGF were protective factors of spastic cerebral palsy [OR (95%CI):2.280 (1.742-2.983),2.222 (1.637-3.016),P<0.05],and NGF was risk factor of spastic cerebral palsy [OR (95%CI):0.720 (0.527-0.984),P＜0.05]. Conclusions The serum expression levels of miR-138 and miR-338 in children with spastic cerebral palsy are significantly up-regulated,and they may play an important role in the occurrence and development of spastic cerebral palsy.
关键词
痉挛型脑性瘫痪 /
血清 /
微小RNA-138 /
微小RNA-338 /
儿童
Key words
spastic cerebral palsy /
serum /
microRNA-138 /
microRNA-338 /
children
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基金
2018年度河南省医学科技攻关计划联合共建项目(2018020192)
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