Objective

This study is to investigate the role of microRNA (miR)-30b in the pathogenesis of hypoxic-ischemic encephalopathy (HIE) in neonates.

Methods

Totally 26 cases of neonatal HIE were included in this study. The protein expression levels of CD26P and PAI-1 were detected with ELISA. Serum levels of miR-30b and PAI-1 mRNA was measured by quantitative real-time PCR. Human brain microvascular endothelial cells (HBMECs) were cultured under hypoxic condition, and the intracellular expression levels of miR-30b and PAI-1 were evaluated. Dual-luciferase reporter assay was performed to confirm the interaction between miR-30b and PAI-1.

Results

Compared with the control group, both the mRNA and protein expression levels of PAI-1 in the serum were up-regulated in the neonates with HIE, together with up-regulated serum CD26P levels. However, the serum expression level of miR-30b was down-regulated in neonatal HIE. In hypoxia-induced HBMECs, the mRNA and protein expression levels of PAI-1 were significantly up-regulated, while the miR-30b expression level was significantly down-regulated. Dual-luciferase reporter assay showed that PAI-1 was the direct target of miR-30b.

Conclusion

Neonatal HIE is accompanied with abnormal platelet activation, significantly up-regulated serum PAI-1 expression levels, and down-regulated miR-30b expression. MiR-30b might regulate the disease pathogenesis and immune responses via modulating PAI-1.

中文翻译：

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MicroRNA-30b对新生儿缺氧缺血性脑病（HIE）的调控机制

目的

这项研究旨在调查microRNA（miR）-30b在新生儿缺氧缺血性脑病（HIE）发病机理中的作用。

方法

本研究共纳入26例新生儿HIE。用ELISA检测CD26P和PAI-1的蛋白表达水平。通过定量实时PCR测量血清miR-30b和PAI-1 mRNA的水平。在缺氧条件下培养人脑微血管内皮细胞（HBMEC），并评估miR-30b和PAI-1的细胞内表达水平。进行双荧光素酶报告基因测定以确认miR-30b和PAI-1之间的相互作用。

结果

与对照组相比，HIE新生儿血清中PAI-1的mRNA和蛋白表达水平均升高，血清CD26P水平也升高。然而，在新生儿HIE中miR-30b的血清表达水平被下调。在缺氧诱导的HBMEC中，PAI-1的mRNA和蛋白表达水平显着上调，而miR-30b的表达水平显着下调。双荧光素酶报告基因检测表明，PAI-1是miR-30b的直接靶标。

结论

新生儿HIE伴有异常的血小板活化，血清PAI-1表达水平明显上调和miR-30b表达下调。MiR-30b可能通过调节PAI-1调节疾病的发病机制和免疫反应。