Gao Jinglin, Wang Huijuan, Zhou Guang, Zhou Zhou, Liang Caixia, Zhong Xiaoning
Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Guangxi Medical University, Nanning 530021, China.
Liuzhou Key Laboratory of Prevention and Treatment of Rheumatic Diseases, The Fourth Affiliated Hospital of Guangxi Medical University, Liuzhou 545005, China.
Mediators Inflamm. 2025 Sep 4;2025:5595989. doi: 10.1155/mi/5595989. eCollection 2025.
To explore the potential role of the RAGE/JAK/STAT pathway along with the activation of myeloid dendritic cells (mDCs) and B cells induced by cigarette smoke exposure in mice. 57BL/6J mice and RAGEfl/flCD11c-Cre mice were subjected to cigarette smoke for 24 weeks and mated with room air controls. Mice bone marrow-derived dendritic cells (BMDCs) were treated with cigarette smoke extracts (CSEs), CSE with the RAGE inhibitor FPS-ZM1 or CSE with the JAK2 inhibitor AG490. The extent of emphysema in these mice was assessed using the average alveolar lining distance (Lm). Real-time PCR was employed to quantify the mRNA expression levels of RAGE, JAK2, STAT1, STAT3 and STAT5 in lung tissue samples. The levels of IL-6 and IL-1β in mouse serum and BMDC supernatant were quantified using ELISA. Flow cytometry was employed to measure the expression of CD40, CD86, RAGE, p-JAK2, p-STAT1, p-STAT3 and p-STAT5 of lung mDCs and BMDCs in mice. Flow cytometry was employed to identify markers CD69, CD86 and CD138 on pulmonary B cells. Exposing mice to cigarette smoke triggered an exaggerated pulmonary mDCs response and elevated the RAGE/JAK/STAT pathway in both pulmonary mDCs and lung tissue, correlating with enhanced B cells response in lungs. Conditional knockdown of RAGE on dendritic cells (DCs) resulted in a reduction of activity within JAK/STAT pathway, impeded the exaggerated mDCs and B cells responses induced by smoking, down-regulated the serum inflammatory response and mitigated emphysema in cigarette smoke-exposed mice. Within a regulated laboratory setting, BMDCs were activated, leading to the amplification of the RAGE/JAK/STAT pathway in these cells after CSE exposure. FPS-ZM1 and AG490 reduced inflammatory factors in the supernatant and activation of BMDC. In mice, prolonged exposure to cigarette smoke triggers the activation of mDCs by enhancing the RAGE/JAK/STAT pathway. Conditional knockdown of RAGE on DCs can prevent the activation of mDCs and B cells triggered by cigarette smoke, indicating that RAGE could be a potential target for treating smoking-induced emphysema.
为了探究晚期糖基化终末产物受体（RAGE）/Janus激酶（JAK）/信号转导和转录激活因子（STAT）通路以及香烟烟雾暴露诱导的小鼠骨髓来源的树突状细胞（mDCs）和B细胞活化的潜在作用。将C57BL/6J小鼠和RAGEfl/flCD11c-Cre小鼠暴露于香烟烟雾中24周，然后与室内空气对照组小鼠进行交配。用香烟烟雾提取物（CSEs）、添加RAGE抑制剂FPS-ZM1的CSE或添加JAK2抑制剂AG490的CSE处理小鼠骨髓来源的树突状细胞（BMDCs）。使用平均肺泡间隔距离（Lm）评估这些小鼠的肺气肿程度。采用实时定量聚合酶链反应（PCR）来定量肺组织样本中RAGE、JAK2、STAT1、STAT3和STAT5的信使核糖核酸（mRNA）表达水平。使用酶联免疫吸附测定（ELISA）定量小鼠血清和BMDCs上清液中白细胞介素（IL）-6和IL-1β的水平。采用流式细胞术检测小鼠肺mDCs和BMDCs中CD40、CD86、RAGE、磷酸化JAK2（p-JAK2）、磷酸化STAT1（p-STAT1）、磷酸化STAT3（p-STAT3）和磷酸化STAT5（p-STAT5）的表达。采用流式细胞术鉴定肺B细胞上的标志物CD69、CD86和CD138。将小鼠暴露于香烟烟雾中会引发过度的肺mDCs反应，并使肺mDCs和肺组织中的RAGE/JAK/STAT通路升高，这与肺中B细胞反应增强相关。在树突状细胞（DCs）上条件性敲低RAGE会导致JAK/STAT通路活性降低，阻碍吸烟诱导的过度mDCs和B细胞反应，下调血清炎症反应，并减轻香烟烟雾暴露小鼠的肺气肿。在规定的实验室环境中，BMDCs被激活，导致CSE暴露后这些细胞中RAGE/JAK/STAT通路的放大。FPS-ZM1和AG490降低了上清液中的炎症因子并抑制了BMDCs的活化。在小鼠中，长期暴露于香烟烟雾会通过增强RAGE/JAK/STAT通路触发mDCs的活化。在DCs上条件性敲低RAGE可以阻止香烟烟雾触发mDCs和B细胞的活化，这表明RAGE可能是治疗吸烟诱导的肺气肿的潜在靶点。
