Ding Haidong, Wu Rina
Altern Ther Health Med. 2024 Apr 18.
Chronic Obstructive Pulmonary Disease (COPD) is characterized by airflow limitation and inflammation resulting from genetic and environmental factors, notably cigarette smoke. Pyroptosis, a cell death process, is implicated in COPD, but its mechanisms are unclear. SHP2, a phosphatase, modulates inflammatory pathways, suggesting a role in COPD pathogenesis and potential therapeutic avenues.
This study investigates the mechanism by which SHP2 regulates cell pyroptosis in bronchial epithelial cells in COPD patients.
In this prospective study, we employed in vivo and in vitro models to investigate the mechanisms underlying COPD progression. Hematoxylin and eosin (H&E) staining were utilized to assess the morphological changes characteristic of COPD. Electron microscopy enabled precise quantification of pyroptotic bodies to highlight cellular changes associated with COPD pathogenesis. Immunofluorescence analysis facilitated the measurement of protein fluorescence intensity, allowing for the assessment of inflammatory responses within bronchial epithelial cells. Additionally, Western blot analysis was conducted to evaluate the expression levels of key pathway proteins involved in COPD progression.
In the COPD model, lesions worsened in SHP2-KD mice compared to SHP2-NC. Western blot results showed increased p22, p47, p-IRE1α, XBP1, STING, p-TBK1, NLRP3, Caspase1, and IL-1β expression levels in both in vivo and in vitro models. Transmission electron microscopy revealed more pyroptotic bodies in SHP2-KD+CSE than in SHP2-NC+CSE. Immunofluorescence demonstrated significantly higher NLRP3 and GSDMD fluorescence intensities in SHP2-KD+CSE versus SHP2-NC+CSE. Additionally, Western blot analysis indicated increased expression of Bax, Caspase3, Caspase8, and Caspase9 proteins in the in vitro model. No differences were observed between SHP2 NC and SHP2-KD groups without CSE stimulation in immunofluorescence, electron microscopy, and Western blot findings in the cellular model.
SHP2 promotes COPD progression by inducing oxidative stress, endoplasmic reticulum stress, and pyroptosis.
慢性阻塞性肺疾病（COPD）的特征是气流受限和由遗传及环境因素（尤其是香烟烟雾）导致的炎症。细胞焦亡是一种细胞死亡过程，与COPD有关，但其机制尚不清楚。SHP2是一种磷酸酶，可调节炎症途径，提示其在COPD发病机制及潜在治疗途径中发挥作用。
本研究探讨SHP2调节COPD患者支气管上皮细胞焦亡的机制。
在这项前瞻性研究中，我们采用体内和体外模型来研究COPD进展的潜在机制。苏木精和伊红（H&E）染色用于评估COPD的形态学变化特征。电子显微镜能够精确量化细胞焦亡小体，以突出与COPD发病机制相关的细胞变化。免疫荧光分析有助于测量蛋白质荧光强度，从而评估支气管上皮细胞内的炎症反应。此外，进行蛋白质印迹分析以评估参与COPD进展的关键信号通路蛋白的表达水平。
在COPD模型中，与SHP2-NC小鼠相比，SHP2-KD小鼠的病变更严重。蛋白质印迹结果显示，体内和体外模型中p22、p47、p-IRE1α、XBP1、STING、p-TBK1、NLRP3、Caspase1和IL-1β的表达水平均升高。透射电子显微镜显示，SHP2-KD+CSE组比SHP2-NC+CSE组有更多的细胞焦亡小体。免疫荧光显示，SHP2-KD+CSE组的NLRP3和GSDMD荧光强度明显高于SHP2-NC+CSE组。此外，蛋白质印迹分析表明，体外模型中Bax、Caspase3、Caspase8和Caspase9蛋白的表达增加。在细胞模型中，未用CSE刺激时，SHP2 NC组和SHP2-KD组在免疫荧光、电子显微镜和蛋白质印迹结果方面未观察到差异。
SHP2通过诱导氧化应激、内质网应激和细胞焦亡促进COPD进展。
