Ma Xin-Gang, Liu Ye, Xue Ming-Xi
Department of Anesthesiology, Zibo Maternal and Child Health Hospital, Zibo 255000, China.
Sheng Li Xue Bao. 2021 Apr 25;73(2):253-262.
The aim of this study was to investigate the effects of dexmedetomidine (Dex) on hepatic ischemia/reperfusion injury (HIRI) and the underlying mechanism. The in vitro HIRI was induced by culturing HL-7702 cells, a human hepatocyte cell line, under 24 h of hypoxia and 12 h of reoxygenation. Quantitative real time PCR (qRT-PCR) and Western blot were performed to detect the expression levels of long non-coding RNA MALAT1, microRNA-126-5p (miR-126-5p) and high mobility group box-1 (HMGB1). Bioinformatics prediction and double luciferase assay were used to verify the targeting relationship between miR-126-5p and MALAT1, HMGB1. Reactive oxygen species (ROS), malondialdehyde (MDA) and ATP levels in culture medium were detected by corresponding kits. The results showed that Dex significantly reduced the levels of ROS and MDA, but increased the level of ATP in HL-7702 cells with HIRI. HIRI up-regulated the expression levels of MALAT1 and HMGB1, and down-regulated the level of miR-126-5p. Dex reversed these effects of HIRI. Furthermore, Dex inhibited HIRI-induced cellular apoptosis, whereas MALAT1 reversed the effect of Dex. This inhibitory effect of Dex could be restored by up-regulation of miR-126-5p. The results suggest that Dex protects hepatocytes from HIRI via regulating MALAT1/miR-126-5p/HMGB1 axis.
本研究旨在探讨右美托咪定（Dex）对肝缺血/再灌注损伤（HIRI）的影响及其潜在机制。通过对人肝细胞系HL-7702细胞进行24小时缺氧培养和12小时复氧来诱导体外HIRI。采用定量实时聚合酶链反应（qRT-PCR）和蛋白质免疫印迹法检测长链非编码RNA MALAT1、微小RNA-126-5p（miR-126-5p）和高迁移率族蛋白B1（HMGB1）的表达水平。利用生物信息学预测和双荧光素酶报告基因检测验证miR-126-5p与MALAT1、HMGB1之间的靶向关系。用相应试剂盒检测培养基中活性氧（ROS）、丙二醛（MDA）和三磷酸腺苷（ATP）水平。结果显示，Dex显著降低了HIRI的HL-7702细胞中ROS和MDA水平，但提高了ATP水平。HIRI上调了MALAT1和HMGB1的表达水平，下调了miR-126-5p水平。Dex逆转了HIRI的这些作用。此外，Dex抑制了HIRI诱导的细胞凋亡，而MALAT1逆转了Dex的作用。上调miR-126-5p可恢复Dex的这种抑制作用。结果表明，Dex通过调节MALAT1/miR-126-5p/HMGB1轴保护肝细胞免受HIRI损伤。
