Clinical value of routine electroencephalogram combined with serum miR-146a and miR-129-5p levels in diagnosis of drug-resistant epilepsy patients

doi:10.11958/20240666

Abstract

Abstract:

Objective To investigate the clinical diagnostic value of routine electroencephalogram (EEG) combined with serum miR-146a and miR-129-5p levels in drug-resistant epilepsy. Methods Sixty patients with refractory epilepsy admitted to our hospital from June 2021 to June 2023 were included in the refractory epilepsy group, and 40 healthy volunteers were included in the control group. Human microvascular endothelial drug-resistant cells (HBMECs) continuously exposed to PHT2 were cultured in vitro and transfected with miR-NC (the miR-NC group), miR-146a mimics (the miR-146a mimics group) and miR-129-5p mimics (the miR-129-5p mimics group), respectively. Western blod assay was used to detect the expression of high-mobility group protein B1 (HMGB1) in each group. Serum miR-146a and miR-129-5p levels were detected by fluorescent quantitative PCR (polymerase chain reaction), and serum HMGB1 protein expression was detected by Western blod assay. The sensitivity, specificity and accuracy of routine EEG, serum miR-146a and miR-129-5p levels and combined diagnosis of drug-resistant epilepsy were evaluated using the comprehensive consultation results of several expert physicians as the gold standard, and ROC curves were drawn. Results Compared with the control group, the expression of HMGB1 was significantly decreased in the drug-resistant group. Compared with the miR-NC group, HMGB1 expression was significantly decreased in the miR-129-5p mimics group and the miR-146a mimics group (P<0.05). Compared with the healthy group, the serum HMGB1 protein expression was down-regulated and miR-146a and miR-129-5p expression levels were significantly up-regulated in the refractory epilepsy group. ROC curve analysis indicated that the area under the curve, sensitivity, specificity and accuracy of conventional EEG combined with serum miR-146a and miR-129-5p levels were higher in the diagnosis of refractory epilepsy than that of single diagnostic method. Conclusion The combination of routine EEG and serum miR-129-5p and miR-146a levels can provide help for the diagnosis of drug-resistant patients.

Key words: drug resistant epilepsy, electroencephalography, microRNAs, ROC curve, HMGB1 protein, miR-146a, miR-129p, drug refractory epilepsy

CLC Number:

R742.1

Figures/Tables 7

Tab.1 Primer sequences for miR-129-5p, miR-146a and U6

基因名称	引物序列（5′→3′）	产物大小/bp
miR-129-5p	上游：CTTTTTGCGGTCTGGGCTTGC	65
	下游：AGTGCAGGGTCCGAGGTATT	65
miR-146a	上游：TGTTGCTGAAGAGCTCTGGTAC	70
	下游：TCGTCTGATGATGTCGAATGTAC	70
U6	上游：CTCGCAACATGGTCGCACAT	66
	下游：GAACGGATCAGATATTGCGT	66

Fig.1 Comparison of HMGB1 protein expression between five groups of cells

Fig.2 Comparison of serum HMGB1 protein expression between the two groups

Tab.2 Comparison of serum levels of miR-129-5p and miR-146a between the two groups of patients

组别	n	miR-129-5p	miR-146a
对照组	40	0.98±0.19	1.13±0.40
难治性癫痫组	60	1.59±0.32	1.96±0.40
t		10.950^＊＊	10.160^＊＊

Fig. 3 Correlation analysis of serum miR-129-5p and miR-146a levels in patients with drug-resistant epilepsy

Tab.3 The diagnostic value of serum miR-129-5p, miR-146a levels, routine EEG and their combined detection in drug-resistant epilepsy

诊断方法	临界值	AUC	95%CI	敏感度/%	特异度/ %	准确度/ %
常规脑电图	-	0.900	0.830~0.970	91.67	82.50	88.00
血清miR-146a	1.54	0.920	0.869~0.971	88.33	85.00	87.00
血清miR-129-5p	1.12	0.930	0.879~0.981	91.67	87.50	90.00
联合诊断	-	0.946	0.891~1.000	96.67	90.00	94.00

Fig.4 ROC curve analysis of serum miR-129-5p, miR-146a levels, routine EEG and combined detection for clinical diagnosis of drug-resistant epilepsy

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