血清miR-146a/b实时荧光定量分析及其作为分子标志物的初步评价

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Abstract Objective: To establish method for the quantitative fluorescence detection of serum miRNAs, and to preliminarily evaluate the potential value of miR-146a/b as serum inflammatory biomarkers. Using serum miR-146a/b, specifically regulating the Toll-like receptor signalling pathway, as the targets. Methods: Serum samples were collected from 20 cases of normal weight children (control group), 20 cases of overweight children (overweight), 20 cases of obese children (obese group) and 50 cases of healthy adults (healthy adult group). Serum total RNAs were extracted by phenol-chloroform extracting methods, copy numbers of serum miR-146a/b were analyzed by SYBR Green real-time fluorescent quantitative PCR techniques, followed by statistical analysis using Graphpad Prism5.0 software and graphics. Results: Copy numbers of serum miR-146a in control group were significantly lower than that in overweight (P=0.0061) and obese groups (P=0.0262), whereas no significant difference was found between overweight group and obesity group (P=0.0656). Area under the ROC curves (AUC) of miR-146a expression in different groups showed as follows: control group vs overweight group AUC=0.8475, P=0.0002; control group vs obese group AUC=0.6050, P=0.2560; overweight group vs obese group AUC=0.5475, P=0.6073. Expression of miR-146b and miR-146a exhibited different trends, copy numbers of serum miR-146b in overweight children were significantly higher than that of normal weight children (P=0.0090) and obese children (P=0.0023); even though the copy numbers of serum miR-146b in obese children were slightly lower than that in control group, the difference had no statistically significant (P=0.1556). The area under the ROC curves (AUC) of miR-146b expression were analyzed as shown in following data: normal group vs overweight group AUC=0.7425, P=0.0087; normal group vs obese group AUC=0.6325, P=0.1517; overweight group vs obese group AUC=0.7825, P=0.0023. The coefficient of variation (CV) values of miR-146a/b copy numbers was largely diverse: CV values of miR-146a copy numbers in control, overweight and obese children were 80.94%, 110.94% and 175.88%, respectively; and that of miR-146b copy numbers were 164.11%, 189.72% and 152.00%, respectively. In healthy adults, the expression patterns of serum miR-146a/b were similar; CV values of miR-146a and miR-146b were 37.86% and 74.82%. Conlusion: The different expression levels of miR-146a in control, overweight and obese groups indicate that miR-146a is correlated with childhood obesity and positively correlated with levels of inflammation in the body. However, the reference range of miR-146a could not be determined on account of its widely varied serum copy numbers and no clear dividing line among different groups. Therefore, the difference of serum miR-146a/b copy number is insufficient to be used in clinical diagnosis, the values of serum miR-146a/b as biomarkers of inflammation still need to be explored further.

Key words： obese children inflammation circulation miRNAs molecular biomarkers

Received: 15 April 2013

Corresponding Authors: 1.School of Laboratory Medicine and Life Sciences, Zhejiang Provincial Key Laboratory of Medical Genetics, Wenzhou Medical University, Wenzhou, 325035; 2.Department of Endocrinology, the Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325027

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	LI Chunmei1
	ZHENG Chao2
	QIAO Minmin1
	LV Jianxin1
	LI Wei1.

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https://xb.wmu.edu.cn/EN/ OR https://xb.wmu.edu.cn/EN/Y2014/V44/I1/1

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