Zeng Qi, Xiao Yan, Zeng Xueliang, Xiao Hai
Department of Ultrasound, First Affiliated Hospital of Gannan Medical University, Ganzhou, China.
Key Laboratory of Prevention and Treatment of Cardiovascular and Cerebrovascular Diseases of Ministry of Education, Gannan Medical University, Ganzhou, China.
PeerJ. 2024 Dec 23;12:e18754. doi: 10.7717/peerj.18754. eCollection 2024.
Ischemic stroke (IS) remains a leading cause of disability and mortality worldwide, and inflammation and oxidative stress play significant roles in its pathogenesis. This study investigates the effects of dihydromyricetin (DHM) on IS using RT-qPCR and western blot with SH-SY5Y cells, focusing on its effects on the small nucleolar RNA host gene 10 (SNHG10)/microRNA (miR)-665/Ras association domain family member 5 (RASSF5) axis and nuclear factor-kappa B (NF-κB) signaling. In addition, the effects of the SNHG10/miR-665/RASSF5 axis on SH-SY5Y cell activity, apoptosis, oxidative stress, and inflammatory markers were analyzed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, flow cytometry, and enzyme-linked immunosorbent assays. Our results showed that, in response to oxygen-glucose deprivation/reperfusion (OGD/R), DHM treatment improved cell viability, reduced apoptosis, and attenuated neuroinflammation and oxidative stress in a dose-dependent manner ( < 0.05). Interestingly, lncRNA SNHG10 was overexpressed during OGD/R and suppressed by DHM. Through bioinformatics analysis and experimental validation, we identified miR-665 as a direct target of SNHG10 and RASSF5 as a direct target of miR-665. The protective effect of DHM against OGD/R injury was partially reversed by SNHG10 overexpression and further enhanced by co-transfection with the miR-665 mimic and si-RASSF5 ( < 0.05). This study identifies a novel mechanism of DHM against IS, which may act via modulation of the SNHG10/miR-665/RASSF5 axis and inactivation of NF-κB signaling, and offers a promising therapeutic target for IS.
缺血性中风（IS）仍然是全球致残和致死的主要原因，炎症和氧化应激在其发病机制中起重要作用。本研究使用RT-qPCR和蛋白质免疫印迹法，以SH-SY5Y细胞为研究对象，探讨二氢杨梅素（DHM）对缺血性中风的影响，重点关注其对小核仁RNA宿主基因10（SNHG10）/微小RNA（miR）-665/Ras关联结构域家族成员5（RASSF5）轴和核因子-κB（NF-κB）信号传导的影响。此外，使用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐、流式细胞术和酶联免疫吸附测定法，分析SNHG10/miR-665/RASSF5轴对SH-SY5Y细胞活性、凋亡、氧化应激和炎症标志物的影响。我们的结果表明，在氧糖剥夺/再灌注（OGD/R）条件下，DHM处理以剂量依赖性方式提高细胞活力、减少凋亡，并减轻神经炎症和氧化应激（P<0.05）。有趣的是，lncRNA SNHG10在OGD/R期间过表达，并被DHM抑制。通过生物信息学分析和实验验证，我们确定miR-665是SNHG10的直接靶点，RASSF5是miR-665的直接靶点。SNHG10过表达部分逆转了DHM对OGD/R损伤的保护作用，而与miR-665模拟物和si-RASSF5共转染进一步增强了这种保护作用（P<0.05）。本研究确定了DHM抗缺血性中风的新机制，其可能通过调节SNHG10/miR-665/RASSF5轴和使NF-κB信号失活发挥作用，并为缺血性中风提供了一个有前景的治疗靶点。
