Su Yinghao, Geng Limei, Ma Yunlei, Yu Xiangyan, Kang Ziyi, Kang Zenglu
Department of Respiratory and Critical Care Medicine, Hebei Provincial Hospital of Traditional Chinese Medicine, Shijiazhuang, Hebei Province, China.
Exp Lung Res. 2021 Oct;47(8):368-381. doi: 10.1080/01902148.2021.1974125. Epub 2021 Sep 11.
House dust mite has been well documented as a major source of allergen in asthma. Circular RNAs (circRNAs) vacuolar protein sorting 33A (circVPS33A, circ_0000455) is overexpressed in a murine asthma model. Herein, we sought to identify its critical action in peptidase 1 (Der p1)-induced dysfunction of BEAS-2B cells.
The levels of circVPS33A, microRNA (miR)-192-5p, and high-mobility group box 1 (HMGB1) were assessed by quantitative real-time PCR (qRT-PCR) or western blot. Actinomycin D treatment and Ribonuclease R (RNase R) assay were used to characterize circVPS33A. Cell viability, proliferation, apoptosis, migration, and invasion were evaluated by Cell Counting Kit-8 (CCK-8), 5-ethynyl-2'-deoxyuridine (EdU), flow cytometry, and transwell assays, respectively. Enzyme-linked immunosorbent assay (ELISA) was used to quantify interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and IL-6. Direct relationship between miR-192-5p and circVPS33A or HMGB1 was verified by dual-luciferase reporter and RNA immunoprecipitation (RIP) assay.
CircVPS33A was highly expressed in asthma plasma and Der p1-treated BEAS-2B cells. Knocking down circVPS33A suppressed Der p1-induced injury in BEAS-2B cells. CircVPS33A targeted miR-192-5p. MiR-192-5p directly targeted HMGB1, and miR-192-5p-mediated repression of HMGB1 alleviated Der p1-driven cell injury. Furthermore, circVPS33A modulated HMGB1 expression through miR-192-5p.
Our findings demonstrated that circVPS33A regulated house dust mite-induced injury in human bronchial epithelial cells at least partially depending on the modulation of the miR-192-5p/HMGB1 axis.
屋尘螨已被充分证明是哮喘中变应原的主要来源。环状RNA（circRNA）空泡蛋白分选33A（circVPS33A，circ_0000455）在小鼠哮喘模型中过表达。在此，我们试图确定其在蛋白酶1（Der p1）诱导的BEAS-2B细胞功能障碍中的关键作用。
通过定量实时PCR（qRT-PCR）或蛋白质免疫印迹法评估circVPS33A、微小RNA（miR）-192-5p和高迁移率族蛋白B1（HMGB1）的水平。放线菌素D处理和核糖核酸酶R（RNase R）检测用于鉴定circVPS33A。分别通过细胞计数试剂盒-8（CCK-8）、5-乙炔基-2'-脱氧尿苷（EdU）、流式细胞术和Transwell检测评估细胞活力、增殖、凋亡、迁移和侵袭。酶联免疫吸附测定（ELISA）用于定量白细胞介素-1β（IL-1β）、肿瘤坏死因子-α（TNF-α）和IL-6。通过双荧光素酶报告基因和RNA免疫沉淀（RIP）检测验证miR-192-5p与circVPS33A或HMGB1之间的直接关系。
circVPS33A在哮喘血浆和Der p1处理的BEAS-2B细胞中高表达。敲低circVPS33A可抑制Der p1诱导的BEAS-2B细胞损伤。circVPS33A靶向miR-192-5p。miR-192-5p直接靶向HMGB1，且miR-192-5p介导的HMGB1抑制减轻了Der p1驱动的细胞损伤。此外，circVPS33A通过miR-192-5p调节HMGB1表达。
我们的研究结果表明，circVPS33A至少部分通过调节miR-192-5p/HMGB1轴来调控屋尘螨诱导的人支气管上皮细胞损伤。
