Zhang Xiao-Yu, Chen Zhuo-Chang, Li Nan, Wang Zhi-Hua, Guo Ya-Li, Tian Cui-Jie, Cheng Dong-Jun, Tang Xue-Yi, Zhang Luo-Xian
Department of Respiratory Disease and Intensive Care, Henan Provincial People's Hospital, Zhengzhou 450003, People's Republic of China.
Department of Respiratory Disease and Intensive Care, People's Hospital Affiliated to Zhengzhou University, Zhengzhou 450003, People's Republic of China.
Hum Mol Genet. 2022 Feb 21;31(4):638-650. doi: 10.1093/hmg/ddab283.
Activated neutrophil-derived exosomes reportedly contribute to the proliferation of airway smooth muscle cells (ASMCs), thereby aggravating the airway wall remodeling during asthma; however, the specific mechanism remains unclear. Lipopolysaccharide (LPS)-EXO and si-CRNDE-EXO were extracted from the media of human neutrophils treated with LPS and LPS + si-CRNDE (a siRNA targets long non-coding RNA CRNDE), respectively. Human ASMCs were co-cultured with LPS-EXO or si-CRNDE-EXO, and cell viability, proliferation and migration were measured. The interplay of colorectal neoplasia differentially expressed (CRNDE), inhibitor of nuclear factor kappa B kinase subunit beta (IKKβ) and nuclear receptor subfamily 2 group C member 2 (TAK1) was explored using RNA immunoprecipitation (RIP) and Co-IP assays. A mouse model of asthma was induced using ovalbumin. CRNDE was upregulated in LPS-EXO and successfully transferred from LPS-treated neutrophils to ASMCs through exosome. Mechanically, CRNDE loaded in LPS-EXO reinforced TAK1-mediated IKKβ phosphorylation, thereby activating the nuclear factor kappa B (NF-κB) pathway. Functionally, silencing CRNDE in LPS-EXO, an IKKβ inhibitor, and an NF-κB inhibitor all removed the upregulation of cell viability, proliferation and migration induced by LPS-EXO in ASMCs. In the end, the in vivo experiment demonstrated that CRNDE knockdown in neutrophils effectively reduced the thickness of bronchial smooth muscle in a mouse model for asthma. Activated neutrophils-derived CRNDE was transferred to ASMCs through exosomes and activated the NF-κB pathway by enhancing IKKβ phosphorylation. The latter promoted the proliferation and migration of ASMCs and then contributed to airway remodeling in asthma.
据报道，活化的中性粒细胞衍生的外泌体有助于气道平滑肌细胞（ASMCs）的增殖，从而加剧哮喘期间的气道壁重塑；然而，具体机制仍不清楚。脂多糖（LPS）-EXO和si-CRNDE-EXO分别从用LPS和LPS+si-CRNDE（一种靶向长链非编码RNA CRNDE的小干扰RNA）处理的人中性粒细胞培养基中提取。将人ASMCs与LPS-EXO或si-CRNDE-EXO共培养，并测量细胞活力、增殖和迁移。使用RNA免疫沉淀（RIP）和免疫共沉淀（Co-IP）实验探究结直肠癌差异表达基因（CRNDE）、核因子κB激酶亚基β（IKKβ）抑制剂和转化生长因子β激活激酶1（TAK1）之间的相互作用。用卵清蛋白诱导建立哮喘小鼠模型。CRNDE在LPS-EXO中上调，并通过外泌体成功地从LPS处理的中性粒细胞转移到ASMCs。从机制上讲，LPS-EXO中携带的CRNDE增强了TAK1介导的IKKβ磷酸化，从而激活核因子κB（NF-κB）通路。在功能上，沉默LPS-EXO中的CRNDE、一种IKKβ抑制剂和一种NF-κB抑制剂均消除了LPS-EXO诱导的ASMCs细胞活力、增殖和迁移的上调。最后，体内实验表明，敲低中性粒细胞中的CRNDE可有效降低哮喘小鼠模型中支气管平滑肌的厚度。活化的中性粒细胞衍生的CRNDE通过外泌体转移到ASMCs，并通过增强IKKβ磷酸化激活NF-κB通路。后者促进了ASMCs的增殖和迁移，进而导致哮喘中的气道重塑。
