丙泊酚与 circAPBB2 协同作用，可防止人心肌细胞缺氧/复氧诱导的氧化应激、炎症和细胞凋亡。

Propofol synergizes with circAPBB2 to protect against hypoxia/reoxygenation-induced oxidative stress, inflammation, and apoptosis of human cardiomyocytes.

机构信息

Department of Anesthesiology, Beijing Tongren Hospital, Capital Medical University, Beijing, China.

Laboratory Medicine, The Affiliated Hospital of Yanbian University, Jilin, China.

出版信息

Immun Inflamm Dis. 2023 Aug;11(8):e952. doi: 10.1002/iid3.952.

DOI:10.1002/iid3.952

PMID:37647434

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10408373/

Abstract

BACKGROUND

Myocardial injury is the main manifestation of cardiovascular diseases, and previous studies have shown that propofol (PPF) regulates myocardial injury. However, the mechanism of PPF in regulating myocardial injury remains to be further explored. This work aims to analyze the effects of PPF on human cardiomyocyte injury and the underlying mechanism.

METHODS

The regulatory and functional role of PPF and circAPBB2 in human cardiomyocyte injury were analyzed using an in vitro hypoxia/reoxygenation (H/R) cell model, which was established by treating human cardiomyocytes (AC16 cells) with H/R. The study evaluated AC16 cell injury by analyzing cytotoxicity, oxidative stress, inflammation and apoptosis of H/R-induced AC16 cells. Quantitative real-time polymerase chain reaction was performed to detect circAPBB2, miR-18a-5p and dual specificity phosphatase 14 (DUSP14) expression. Protein expression was analyzed by Western blot analysis assay. Dual-luciferase reporter assay, RNA pull-down assay and RNA immunoprecipitation assay were performed to identify the associations among circAPBB2, miR-18a-5p and DUSP14. Cytotoxicity was investigated by cell counting kit-8 assay and lactate dehydrogenase activity detection kit. Oxidative stress was evaluated by cellular reactive oxygen species assay kit and superoxide dismutase activity assay kit. The production of tumor necrosis factor-α and interleukin-1β was evaluated by enzyme-linked immunosorbent assays.

RESULTS

The expression of circAPBB2 and DUSP14 was significantly decreased, while miR-18a-5p was increased in H/R-induced AC16 cells when compared with controls. H/R treatment-induced cytotoxicity, oxidative stress, inflammation and cell apoptosis were attenuated after circAPBB2 overexpression or PPF treatment, whereas these effects were restored by increasing miR-18a-5p expression. PPF treatment improved the inhibitory effect of ectopic circAPBB2 expression on H/R-induced cell injury. MiR-18a-5p silencing ameliorated H/R-induced AC16 damage by interacting with DUSP14. Mechanically, circAPBB2 acted as a miR-18a-5p sponge, and miR-18a-5p targeted DUSP14 in AC16 cells.

CONCLUSION

PPF synergized with circAPBB2 to protect AC16 cells against H/R-induced oxidative stress, inflammation and apoptosis through the miR-18a-5p/DUSP14 pathway.

摘要

背景

心肌损伤是心血管疾病的主要表现，先前的研究表明丙泊酚（PPF）可调节心肌损伤。然而，PPF 调节心肌损伤的机制仍有待进一步探讨。本研究旨在分析 PPF 对人心肌细胞损伤的影响及其潜在机制。

方法

采用体外缺氧/复氧（H/R）细胞模型分析 PPF 和 circAPBB2 在人心肌细胞（AC16 细胞）损伤中的调节和功能作用，该模型通过用 H/R 处理人心肌细胞（AC16 细胞）建立。通过分析 H/R 诱导的 AC16 细胞的细胞毒性、氧化应激、炎症和细胞凋亡来评估 AC16 细胞损伤。采用实时定量聚合酶链反应检测 circAPBB2、miR-18a-5p 和双特异性磷酸酶 14（DUSP14）的表达。通过 Western blot 分析测定蛋白表达。通过双荧光素酶报告基因检测、RNA 下拉测定和 RNA 免疫沉淀测定鉴定 circAPBB2、miR-18a-5p 和 DUSP14 之间的关联。通过细胞计数试剂盒-8 检测和乳酸脱氢酶活性检测试剂盒检测细胞毒性。通过细胞内活性氧检测试剂盒和超氧化物歧化酶活性检测试剂盒评估氧化应激。通过酶联免疫吸附测定评估肿瘤坏死因子-α和白细胞介素-1β的产生。

结果

与对照组相比，H/R 诱导的 AC16 细胞中 circAPBB2 和 DUSP14 的表达显著降低，而 miR-18a-5p 的表达增加。过表达 circAPBB2 或 PPF 处理后，H/R 诱导的细胞毒性、氧化应激、炎症和细胞凋亡减弱，而增加 miR-18a-5p 的表达则恢复了这些作用。PPF 处理改善了外源性 circAPBB2 表达对 H/R 诱导的细胞损伤的抑制作用。沉默 miR-18a-5p 可通过与 DUSP14 相互作用改善 H/R 诱导的 AC16 损伤。机制上，circAPBB2 作为 miR-18a-5p 的海绵，miR-18a-5p 在 AC16 细胞中靶向 DUSP14。