摘要
目的探讨内质网应激(ERS)信号通路在哮喘小鼠气道黏液高分泌及气道重塑中的作用。方法选择无特定病原体(SPF)级雌性C57BL/6小鼠24只,实验分为对照组、模型组、ERS抑制剂组,每组8只。除对照组外,其余两组卵清清蛋白(OVA)+氢氧化铝制备哮喘模型,从第21天起ERS抑制剂组每次激发前30 min灌胃0.35 mg/g 4-苯基丁酸(4-PBA),对照组和模型组灌胃等体积生理盐水。对肺泡灌洗液(BALF)中细胞计数;酶联免疫吸附试验(ELISA)检测BALF中白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)水平;苏木精-伊红(HE)染色检测肺组织形态情况;马松染色检测肺组织胶原沉积情况;过碘酸雪夫(PAS)染色检测肺组织杯状上皮细胞增生情况;Western blot检测肺组织中肌醇需求酶1α(IRE1α)、活化转录因子6(ATF6)、ERS相关基因C/EBP同源蛋白(CHOP)、葡萄糖调节蛋白78(GRP78)表达情况。结果相较于对照组,模型组肺血管周围聚集大量炎症细胞、肺泡间隔增厚、结构破坏、平滑肌增厚,胶原纤维沉积明显增加,杯状细胞数量明显增多;ERS抑制剂组相较于模型组炎症细胞数量减少、肺泡间隔有所增加,但结构破坏仍严重,胶原纤维沉积、杯状细胞数量均降低。与对照组比较,模型组BALF中细胞总数、嗜酸粒细胞、中性粒细胞、淋巴细胞、巨噬细胞数量,IL-6、TNF-α水平,肺组织中IRE1α、ATF6、CHOP、GRP78水平均升高(P<0.05);与模型组比较,ERS抑制剂组BALF中细胞总数、嗜酸粒细胞、中性粒细胞、淋巴细胞、巨噬细胞数量,IL-6、TNF-α水平,肺组织中IRE1α、ATF6、CHOP、GRP78水平均降低(P<0.05)。结论抑制ERS信号通路可以抑制炎性反应和气道重塑,缓解气道黏液高分泌情况,实现对哮喘小鼠的保护。
Objective To investigate the role of endoplasmic reticulum stress(ERS)signaling pathway in airway mucus hypersecretion and airway remodeling of asthmatic mice.Methods Twenty-four specific pathogen-free(SPF)grade female C57BL/6 mice were selected and divided into the control group,model group and ERS inhibitor group,8 cases in each group.Except for the control group,the asthma model was prepared by ovalbumin(OVA)and aluminum hydroxide in the other two groups,then from 21 d,the ERS inhibitor group was given 0.35 mg/g of 4-phenylbutyric acid(4-PBA)by gavage at 30 min before each stimulation,and the control group and the model group were given the same volume of normal saline by gavage.The cells in bronchial alveolar lavage fluid(BALF)were counted;the levels of interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in BALF were detected by the enzyme-linked immunosorbent assay(ELISA);the morphology of lung tissues was detected by hematoxylin eosin(HE)staining;the collagen deposition in lungtissues was detected by Masson staining;the proliferation of goblet epithelial cells was detected by Periodic Acid-Schiff(PAS)staining;in addition,the protein expressions of inositol-requiring enzyme 1α(IRE1α),activating transcription factor 6(ATF6),endoplasmic reticulum stress-related gene C/EBP homologous protein(CHOP)and glucose-regulated protein 78(GRP78)in lung tissues were detected by Western blot.Results In the model group,a large number of inflammatory cells gathered around the pulmonary vessels,the alveolar septum was thickened,the structure was destroyed,and smooth muscle was thickened,the collagen deposition was increased significantly,and the number of goblet cells was increased significantly;compared with those in the model group,the number of inflammatory cells in the ERS inhibitor group was decreased,the alveolar septum was widened,but the structural damage was still serious,the collagen fiber deposition and the number of goblet cells were decreased.Compared with those in the control group,the total number of cells
作者
陈家亮
周向东
陈小妹
李琪
刘锋
CHEN Jialiang;ZHOU Xiangdong;CHEN Xiaomei;LI Qi;LIU Feng(Department of Respiratory Medicine,First Affiliated Hospital of Hainan Medical University,Haikou,Hainan 570102,China;Key Laboratory of Emergency and Trauma,Ministry of Education,Haikou,Hainan 571199,China;Department of General Medicine,First Affiliated Hospital of Hainan Medical University,Haikou,Hainan 570102,China)
出处
《重庆医学》
CAS
2021年第10期1633-1637,共5页
Chongqing medicine
基金
国家自然科学基金项目(81811530063)
海南医学院第一附属医院青年培育基金项目(HYFYPY201808)。
关键词
内质网应激信号通路
哮喘
小鼠
气道黏液高分泌
气道重塑
endoplasmic reticulum stress signaling pathway
asthma
mice
airway mucus hypersecretion
airway remodeling
