Shi Lu, Huang Man, Chen Si-An, Xu Jun-Peng, Zhang Qi-Hao, Cao Wen-Jie, Tian Yun-Na, Wang Xiao-Ting, Wang Wan-Tie
College of Physical Education and Health, Yibin University, Yibin 644000, China.
Institute of Ischemia/Reperfusion Injury, Wenzhou Medical University, Wenzhou 325035, China.
Sheng Li Xue Bao. 2024 Aug 25;76(4):517-525.
The aim of this study was to investigate whether the protective effect of 2-deoxyglucose (2-DG) on lung ischemia/reperfusion (I/R) injury is mediated by inhibiting nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3)-mediated pyroptosis in rats. Male Sprague-Dawley rats were randomly divided into control group, 2-DG group, lung I/R injury group (I/R group) and 2-DG+I/R group. 2-DG (0.7 g/kg) was intraperitoneally injected 1 h prior to lung ischemia. The tissue structure was measured under light microscope. Lung injury parameters were detected. The contents of malondialdehyde (MDA), myeloperoxidase (MPO) and lactate were determined by commercially available kits. ELISA was used to detect the levels of IL-1β and IL-18. Western blot, qRT-PCR and immunofluorescence staining were used to measure the expression changes of glycolysis and pyroptosis related indicators. The results showed that there was no significant difference in the parameters between the control group and the 2-DG group. However, the lung injury parameters, oxidative stress response, lactic acid content, IL-1β, and IL-18 levels were significantly increased in the I/R group. The protein expression levels of glycolysis and pyroptosis related indicators including hexokinase 2 (HK2), pyruvate kinase 2 (PKM2), NLRP3, Gasdermin superfamily member GSDMD-N, cleaved-Caspase1, cleaved-IL-1β and cleaved-IL-18, and the gene expression levels of HK2, PKM2 and NLRP3 were markedly up-regulated in the I/R group compared with those in the control group. The expression of HK2 and NLRP3 was also increased detected by immunofluorescence staining. Compared with the I/R group, the 2-DG+I/R group exhibited significantly improved alveolar structure and inflammatory infiltration, reduced lung injury parameters, and decreased expression of glycolysis and pyroptosis related indicators. These results suggest that 2-DG protects against lung I/R injury possibly by inhibiting NLRP3-mediated pyroptosis in rats.
本研究旨在探讨2-脱氧葡萄糖（2-DG）对大鼠肺缺血/再灌注（I/R）损伤的保护作用是否通过抑制核苷酸结合寡聚化结构域样受体蛋白3（NLRP3）介导的细胞焦亡来实现。雄性Sprague-Dawley大鼠随机分为对照组、2-DG组、肺I/R损伤组（I/R组）和2-DG+I/R组。在肺缺血前1小时腹腔注射2-DG（0.7 g/kg）。在光学显微镜下测量组织结构。检测肺损伤参数。使用商用试剂盒测定丙二醛（MDA）、髓过氧化物酶（MPO）和乳酸的含量。采用酶联免疫吸附测定（ELISA）法检测白细胞介素-1β（IL-1β）和白细胞介素-18（IL-18）水平。采用蛋白质免疫印迹法（Western blot）、实时荧光定量聚合酶链反应（qRT-PCR）和免疫荧光染色法检测糖酵解和细胞焦亡相关指标的表达变化。结果显示，对照组和2-DG组之间的参数无显著差异。然而，I/R组的肺损伤参数、氧化应激反应、乳酸含量、IL-1β和IL-18水平显著升高。与对照组相比，I/R组中包括己糖激酶2（HK2）﹑丙酮酸激酶2（PKM2）、NLRP3、Gasdermin超家族成员GSDMD-N、裂解的半胱天冬酶-1（cleaved-Caspase1）、裂解的IL-1β和裂解的IL-18等糖酵解和细胞焦亡相关指标的蛋白质表达水平以及HK2、PKM2和NLRP3的基因表达水平均明显上调。免疫荧光染色也检测到HK2和NLRP3表达增加。与I/R组相比，2-DG+I/R组的肺泡结构和炎症浸润明显改善，肺损伤参数降低，糖酵解和细胞焦亡相关指标的表达减少。这些结果表明，2-DG可能通过抑制大鼠NLRP3介导的细胞焦亡来预防肺I/R损伤。
