Department of Anesthesiology, Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hospital (SAMS&SPPH, Affiliated Hospital of UESTC), Chengdu, Sichuan, China.
Hum Gene Ther. 2022 Oct;33(19-20):1073-1090. doi: 10.1089/hum.2022.078. Epub 2022 Aug 22.
Sepsis is a common life-threatening pathology. This study investigated the role of transcription factor sex-determining region Y (SRY)-box 9 (SOX9) in sepsis-induced cardiomyocyte pyroptosis. A murine model of sepsis was established, followed by detection of cardiac functions and myocardial injury. HL-1 mouse cardiomyocytes were induced by lipopolysaccharides (LPS). The levels of interleukin (IL)-18, IL-1β, tumor necrosis factor-α (TNF-α), IL-6, malondialdehyde (MDA), and superoxide dismutase (SOD) in myocardial tissues and HL-1 mouse cardiomyocytes were determined. SOX9 ubiquitination level was measured. The binding relationships between SOX9-miR-96-5p and miR-96-5p-NLR pyrin domain containing 3 (NLRP3) were analyzed, and the interaction between ubiquitin-specific peptidase 7 (USP7) and SOX9 was measured. SOX9 was highly expressed in septic mice and LPS-induced HL-1 mouse cardiomyocytes. SOX9 silencing improved cardiac function, alleviated myocardial injury, reduced the levels of IL-1β, IL-18, cleaved caspase-1, gasdermin D N-terminal domain, TNF-α, IL-6, and MDA in myocardial tissues and HL-1 mouse cardiomyocytes, increased the level of SOD, and alleviated cardiomyocyte pyroptosis. USP7 upregulated SOX9 expression through deubiquitination. SOX9 inhibited miR-96-5p expression and miR-96-5p targeted NLRP3. miR-96-5p silencing or USP7 overexpression reversed the inhibitory effect of SOX9 silencing on cardiomyocyte pyroptosis. Collectively, USP7 upregulated SOX9 expression through deubiquitination, and SOX9 suppressed miR-96-5p expression by binding to the miR-96-5p promoter region, thereby promoting NLRP3 expression and then exacerbating sepsis-induced myocardial injury and cardiomyocyte pyroptosis.
脓毒症是一种常见的危及生命的病理。本研究探讨了转录因子性别决定区 Y（SRY）-盒 9（SOX9）在脓毒症诱导的心肌细胞细胞焦亡中的作用。建立脓毒症小鼠模型，检测心功能和心肌损伤。用脂多糖（LPS）诱导 HL-1 小鼠心肌细胞。检测心肌组织和 HL-1 小鼠心肌细胞中白细胞介素（IL）-18、IL-1β、肿瘤坏死因子-α（TNF-α）、IL-6、丙二醛（MDA）和超氧化物歧化酶（SOD）的水平。测定 SOX9 泛素化水平。分析 SOX9-miR-96-5p 与 miR-96-5p-NLR 吡喃结构域包含 3（NLRP3）的结合关系，并测定泛素特异性肽酶 7（USP7）与 SOX9 的相互作用。SOX9 在脓毒症小鼠和 LPS 诱导的 HL-1 小鼠心肌细胞中高表达。沉默 SOX9 可改善心功能，减轻心肌损伤，降低心肌组织和 HL-1 小鼠心肌细胞中 IL-1β、IL-18、裂解型半胱天冬酶-1、gasdermin D N 端结构域、TNF-α、IL-6、MDA 的水平，增加 SOD 水平，减轻心肌细胞细胞焦亡。USP7 通过去泛素化上调 SOX9 表达。SOX9 抑制 miR-96-5p 的表达，miR-96-5p 靶向 NLRP3。沉默 miR-96-5p 或过表达 USP7 可逆转 SOX9 沉默对心肌细胞细胞焦亡的抑制作用。总之，USP7 通过去泛素化上调 SOX9 表达，SOX9 通过与 miR-96-5p 启动子区结合抑制 miR-96-5p 的表达，从而促进 NLRP3 的表达，进而加重脓毒症引起的心肌损伤和心肌细胞细胞焦亡。
