Zhang Rongfang, Yang Aiping, Fu Jin, Zhang Li, Yin Liyue, Xu Ting, Dai Chunhui, Su Wenbing, Shen Wanling
Department of Rehabilitation Medicine, Qujing No. 1 Hospital, No. 1 Garden Road, Qilin District, Qujing, Yunnan 655000, China.
Toxicol Res (Camb). 2024 Aug 2;13(4):tfae115. doi: 10.1093/toxres/tfae115. eCollection 2024 Aug.
The anti-inflammatory effects of budesonide (BUN) and N-acetylcysteine (NAC) attenuate acute lung injury (ALI). The aim of this study was to investigate the effects of combination therapy consisting of BUN and NAC on ALI and the underlying mechanisms.
In vitro and in vivo models of ALI were generated by LPS induction. Western blotting was used to detect the expression levels of pyroptosis-related proteins and inflammation-related factors, and RT-qPCR was used to detect the expression of miR-381. Cell proliferation and apoptosis were detected by CCK-8 and flow cytometry, respectively. ELISA was used to detect the levels of inflammation-related factors. HE staining was used to detect lung injury.
The results showed that LPS effectively induced pyroptosis in cells and promoted the expression of pyroptosis-related proteins (Caspase1, Gasdermin D and NLRP3) and inflammatory cytokines (TNF-α, IL-6 and IL-1β). The combination of BUN and NAC significantly alleviated LPS-induced pyroptosis and inflammation. In addition, the combination of BUN and NAC effectively promoted miR-381 expression. Transfection of miR-381 mimics effectively alleviated LPS-induced pyroptosis and inflammation, while transfection of miR-381 inhibitors had the opposite effect. miR-381 negatively regulates NLRP3 expression. Treatment with a miR-381 inhibitor or pc-NLRP3 reversed the effects of the combination of BUN and NAC. In a mouse model of ALI, the combination of BUN and NAC effectively improved lung injury, while treatment with a miR-381 inhibitor or pc-NLRP3 effectively reversed this effect.
Overall, this study revealed that BUN + NAC inhibits the activation of NLRP3 by regulating miR-381, thereby alleviating ALI caused by pyroptosis-mediated inflammation.
布地奈德（BUN）和N-乙酰半胱氨酸（NAC）的抗炎作用可减轻急性肺损伤（ALI）。本研究旨在探讨BUN与NAC联合治疗对ALI的影响及其潜在机制。
通过脂多糖（LPS）诱导建立ALI的体外和体内模型。采用蛋白质免疫印迹法检测焦亡相关蛋白和炎症相关因子的表达水平，采用逆转录-定量聚合酶链反应（RT-qPCR）检测miR-381的表达。分别采用细胞计数试剂盒-8（CCK-8）和流式细胞术检测细胞增殖和凋亡。采用酶联免疫吸附测定（ELISA）检测炎症相关因子水平。采用苏木精-伊红（HE）染色检测肺损伤情况。
结果显示，LPS可有效诱导细胞焦亡，促进焦亡相关蛋白（半胱天冬酶1、gasdermin D和NLRP3）和炎性细胞因子（肿瘤坏死因子-α、白细胞介素-6和白细胞介素-1β）的表达。BUN与NAC联合使用可显著减轻LPS诱导的焦亡和炎症反应。此外，BUN与NAC联合使用可有效促进miR-381的表达。转染miR-381模拟物可有效减轻LPS诱导的焦亡和炎症反应，而转染miR-381抑制剂则产生相反的效果。miR-381负向调节NLRP3的表达。用miR-381抑制剂或pc-NLRP3处理可逆转BUN与NAC联合使用的效果。在ALI小鼠模型中，BUN与NAC联合使用可有效改善肺损伤，而用miR-381抑制剂或pc-NLRP3处理可有效逆转这一效果。
总体而言，本研究表明BUN + NAC通过调节miR-381抑制NLRP3的激活，从而减轻由焦亡介导的炎症反应引起的ALI。
