Li Ya-Hong, Zhang Shun, Tang Lu, Feng Jianguo, Jia Jing, Chen Ye, Liu Li, Zhou Jun
Department of Anesthesiology, The Affiliated Hospital of Southwest Medical University, Luzhou 646000, China.
Anesthesiology and Critical Care Medicine Key Laboratory of Luzhou, The Affiliated Hospital of Southwest Medical University, Luzhou 646000, China.
J Pers Med. 2023 Apr 3;13(4):631. doi: 10.3390/jpm13040631.
Cerebral ischemia/reperfusion (CI/R) injury causes high disability and mortality. Hydrogen (H) enhances tolerance to an announced ischemic event; however, the therapeutic targets for the effective treatment of CI/R injury remain uncertain. Long non-coding RNA lincRNA-erythroid prosurvival (EPS) (lincRNA-EPS) regulate various biological processes, but their involvement in the effects of H and their associated underlying mechanisms still needs clarification. Herein, we examine the function of the lincRNA-EPS/Sirt1/autophagy pathway in the neuroprotection of H against CI/R injury. HT22 cells and an oxygen-glucose deprivation/reoxygenation (OGD/R) model were used to mimic CI/R injury in vitro. H, 3-MA (an autophagy inhibitor), and RAPA (an autophagy agonist) were then administered, respectively. Autophagy, neuro-proinflammation, and apoptosis were evaluated by Western blot, enzyme-linked immunosorbent assay, immunofluorescence staining, real-time PCR, and flow cytometry. The results demonstrated that H attenuated HT22 cell injury, which would be confirmed by the improved cell survival rate and decreased levels of lactate dehydrogenase. Furthermore, H remarkably improved cell injury after OGD/R insult via decreasing pro-inflammatory factors, as well as suppressing apoptosis. Intriguingly, the protection of H against neuronal OGD/R injury was abolished by rapamycin. Importantly, the ability of H to promote lincRNA-EPS and Sirt1 expression and inhibit autophagy were abrogated by the siRNA-lincRNA-EPS. Taken together, the findings proved that neuronal cell injury caused by OGD/R is efficiently prevented by H via modulating lincRNA-EPS/Sirt1/autophagy-dependent pathway. It was hinted that lincRNA-EPS might be a potential target for the H treatment of CI/R injury.
脑缺血/再灌注（CI/R）损伤会导致高致残率和死亡率。氢气（H₂）可增强对已知缺血事件的耐受性；然而，CI/R损伤有效治疗的靶点仍不明确。长链非编码RNA lincRNA-红细胞促生存因子（EPS）（lincRNA-EPS）调节多种生物学过程，但其在H₂作用中的参与情况及其相关潜在机制仍需阐明。在此，我们研究lincRNA-EPS/沉默调节蛋白1（Sirt1）/自噬途径在H₂对CI/R损伤的神经保护中的作用。使用HT22细胞和氧糖剥夺/复氧（OGD/R）模型在体外模拟CI/R损伤。然后分别给予H₂、3-甲基腺嘌呤（3-MA，一种自噬抑制剂）和雷帕霉素（RAPA，一种自噬激动剂）。通过蛋白质免疫印迹法、酶联免疫吸附测定、免疫荧光染色、实时聚合酶链反应和流式细胞术评估自噬、神经促炎反应和细胞凋亡。结果表明，H₂减轻了HT22细胞损伤，这可通过提高细胞存活率和降低乳酸脱氢酶水平得到证实。此外，H₂通过降低促炎因子以及抑制细胞凋亡，显著改善了OGD/R损伤后的细胞损伤。有趣的是，雷帕霉素消除了H₂对神经元OGD/R损伤的保护作用。重要的是，小干扰RNA（siRNA）-lincRNA-EPS消除了H₂促进lincRNA-EPS和Sirt1表达以及抑制自噬的能力。综上所述，这些发现证明H₂通过调节lincRNA-EPS/Sirt1/自噬依赖性途径有效预防了OGD/R引起的神经元细胞损伤。这提示lincRNA-EPS可能是H₂治疗CI/R损伤的潜在靶点。
