吉非替尼对哮喘C57BL/6小鼠气道炎症和气道重塑的影响及作用机制

doi:10.3760/cma.j.cn112137-20231205-01297

科研成果详情

题名	吉非替尼对哮喘C57BL/6小鼠气道炎症和气道重塑的影响及作用机制
其他题名	The effects and mechanisms of Gefitinib on airway inflammation and airway remodeling in C57BL/6 mice with asthma
作者	吴立琴1; 郑锐1; 李飞静1; 王美艳 2; 鲁春 3; 方乐彤 1,4; 张亚利1,4; 戴元荣1
发表日期	2024-05-28
发表期刊	中华医学杂志影响因子和分区
语种	中文
原始文献类型	Periodical
关键词	哮喘吉非替尼气道炎症气道重塑小鼠
其他关键词	Asthma ; Gefitinib ; Airway inflammation ; Airway remodeling ; Mice
摘要	目的:探讨表皮生长因子受体（EGFR）抑制剂吉非替尼对哮喘C57BL/6小鼠气道炎症和气道重塑的作用，并分析其作用机制。方法:用随机数字法将雄性6~8周龄C57BL/6小鼠分为A组（对照组）、B组（哮喘组）、C组（哮喘+20 mg/kg吉非替尼组）、D组（哮喘+40 mg/kg吉非替尼组）和E组（40 mg/kg吉非替尼组），每组7只。分别于第0、14天腹腔内注射卵清蛋白（OVA）及Al（OH） 3［20 μg OVA+2 mg Al（OH） 3溶于0.2 ml生理盐水］混合液0.2 ml致敏；B、C、D组分别于第25至31天开始雾化吸入1% OVA溶液8 ml激发哮喘，1次/d，每次约40 min，连续雾化7 d。C、D组每次雾化激发前半小时灌胃给予0.5%羧甲基纤维素钠（CMCNa）溶解的吉非替尼0.2 ml，E组仅同时给予0.5% CMCNa溶解的吉非替尼0.2 ml。A组和B组灌胃给予等量的0.5% CMCNa。末次激发24 h后，收集小鼠支气管肺泡灌洗液（BALF）进行总细胞计数及嗜酸性粒细胞计数，ELISA检测各组小鼠血清总IgE、BALF及肺组织匀浆中白细胞介素（IL）-4、IL-5和IL-13水平；检测肺组织中IL-4、IL-5和IL-13的mRNA表达水平；免疫组化、Western印迹实验检测肺组织中EGFR等指标的表达水平。结果:B组小鼠血清总IgE水平，BALF中总细胞计数、嗜酸性粒细胞计数、IL-4、IL-5、IL-13水平，肺组织中EGFR的磷酸化及下游激活水平均高于A组（均 P<0.05）。C、D组小鼠血清总IgE水平［（261.32±44.38）、（194.09±52.39）比（1 023.70±105.51）ng/ml］，BALF中总细胞计数［（23.70±4.08）×10 5个/ml、（14.92±4.06）×10 5个/ml比（35.36±6.30）×10 5个/ml］、嗜酸性粒细胞计数［（108.00±13.69）×10 4个/ml、（67.00±17.28）×10 4个/ml比（147.86±20.06）×10 4个/ml］，IL-4［（36.42±4.48）、（30.45±8.12）比（58.72±7.17）pg/ml］、IL-5［（16.20±4.62）、（13.38±5.14）比（23.46±5.38）pg/ml］、IL-13［（18.45±7.28）、（14.33±7.70）比（104.12±24.66）pg/ml］水平均低于B组（均 P<0.05）；肺组织中IL-4、IL-5、IL-13水平及其mRNA水平均低于B组（均 P<0.05）。C、D组小鼠肺组织中磷酸化表皮生长因子受体（p-EGFR）阳性表达率［（40.53±6.80）%、（23.60±4.42）%比（70.78±5.36）%］、p-EGFR/EGFR（61.68±7.48、51.13±5.19比105.90±11.66）、磷酸化细胞外调节蛋白激酶（p-Erk）/细胞外调节蛋白激酶（Erk）（75.28±7.11、47.54±4.83比98.76±4.71）和磷酸化蛋白激酶B（p-Akt）/蛋白激酶B（Akt）（96.24±5.40、68.52±2.73比103.30±4.52）比值均低于B组（均 P<0.05）。A、E组相关指标差异均无统计学意义（均 P0.05）。结论:吉非替尼可能通过抑制EGFR磷酸化，影响下游Erk和Akt的活化，进而缓解哮喘小鼠气道炎症和气道重塑。
其他摘要	Objective:To investigate the effects of the epidermal growth factor receptor(EGFR) inhibitor Gefitinib on airway inflammation and airway remodelling in asthmatic C57BL/6 mice, and to analyze its possible mechanisms.Methods:Male C57BL/6 mice, aged 6-8 weeks, were randomly assigned into five groups: Group A (control group), Group B (asthma group), Group C (asthma+20 mg/kg gefitinib group), Group D (asthma+40 mg/kg gefitinib group), and Group E (40 mg/kg gefitinib group), with seven mice per group. Mice were sensitized by intraperitoneal injection of a mixture of 0.2 ml solution containing OVA and Al(OH) 3 [20 μg OVA+2 mg Al(OH) 3 dissolved in 0.2 ml of physiological saline] at Day 0 and 14. Starting from Day 25 to 31, Group B, C, and D were challenged with nebulization of 1% OVA solution (8 ml) to induce asthma, once a day for approximately 40 minutes, with continuous aerosolization for 7 days. Group C and D were given 0.2 ml of Gefitinib dissolved in 0.5% carboxymethylcellulose sodium (CMCNa) by gavage half an hour before challenging, and Group E was simultaneously given with 0.2 ml of Gefitinib dissolved in 0.5% CMCNa only. Group A and B were given an equivalent volume of 0.5% CMCNa by gavage. After 24 h of final challenge, the bronchoalveolar lavage fluid (BALF) was prepared for the determination of total cell count and eosinophil count. The levels of total immune globulin E (IgE) in serum and interleukin (IL)-4, IL-5 and IL-13 in BALF and lung tissue homogenates were measured by ELISA. The mRNA expression levels of IL-4, IL-5, IL-13 in lung were measured. Immunohistochemistry and Western blot experiments were used to detect the expression levels of EGFR in lung tissues. Results:In Group B, the level of total IgE in serum, total cell count, eosinophil count, the levels of IL-4, IL-5, IL-13 in BALF and the phosphorylation of EGFR and its downstream activation in lung were higher than those in Group A (all P<0.05). The levels of total IgE in serum [(261.32±44.38) ng/ml, (194.09±52.39) ng/ml vs (1 023.70±105.51) ng/ml], total cell count [(23.70±4.08)×10 5/ml, (14.92±4.06)×10 5/ml vs (35.36±6.30)×10 5/ml], eosinophil count [(108.00±13.69)×10 4/ml, (67.00±17.28)×10 4/ml vs (147.86±20.06)×10 4/ml], IL-4 [(36.42±4.48) pg/ml, (30.45±8.12) pg/ml vs (58.72±7.17) pg/ml], IL-5 [(16.20±4.62) pg/ml, (13.38±5.14) pg/ml vs (23.46±5.38) pg/ml], IL-13 [(18.45±7.28) pg/ml, (14.33±7.70) pg/ml vs (104.12±24.66) pg/ml] in BALF of Group C and D were lower than those in Group B (all P<0.05). The levels of IL-4, IL-5, and IL-13 as well as their mRNA levels in the lung tissue of Group C and D were lower than those in Group B (all P<0.05). In Group C and D, the positive expression rate of phosphorylated epidermal growth factor receptor (p-EGFR) in lung tissue [(40.53±6.80)%, (23.60±4.42)% vs (70.78±5.36)%], p-EGFR/EGFR (61.68±7.48, 51.13±5.19 vs 105.90±11.66), phosphorylated extracellular regulated protein kinase (p-Erk)/extracellular regulated protein kinase (Erk) (75.28±7.11, 47.54±4.83 vs 98.76±4.71), and phosphorylated protein kinase B (p-Akt)/protein kinase B (Akt) (96.24±5.40, 68.52±2.73 vs 103.30±4.52) was lower than those of Group B (all P<0.05). There was no statistically significant difference in the relevant indicators between Group A and E (all P0.05). Conclusion:Gefitinib may alleviate airway inflammation and airway remodeling in asthmatic mice by inhibiting EGFR phosphorylation and affecting the activation of downstream Erk and Akt.
资助项目	National Natural Science Foundation of China[82173876]
出版者	Chinese Medical Journals Publishing House Co.Ltd
ISSN	0376-2491
卷号	104 期号:20 页码:1860-1867
DOI	10.3760/cma.j.cn112137-20231205-01297
页数	8
收录类别	万方 ; PUBMED ; SCOPUS ; CSCD ; ISTIC ; 北大核心
URL	查看原文
PubMed ID	38782755
SCOPUSEID	2-s2.0-85194125977
通讯作者地址	[Dai, Yuanrong]Department of Pulmonary and Critical Care Medicine,The Second Affiliated Hospital and Yuying Children′s Hospital of Wenzhou Medical University,Wenzhou,325000,China
Scopus学科分类	Medicine (all)
引用统计
文献类型	期刊论文
条目标识符	https://kms.wmu.edu.cn/handle/3ETUA0LF/214819
专题	附属第二医院药学院（分析测试中心）第二临床医学院、附属第二医院、育英儿童医院附属第二医院_重症医学科
作者单位	1.温州医科大学附属第二医院(育英儿童医院)呼吸与危重症医学科，温州　325000; 2.浙江省台州市第一人民医院呼吸与危重症医学科，台州　318000; 3.浙江省萧山区第一人民医院呼吸与危重症医学科，杭州　311200; 4.温州医科大学药学院化学生物学研究中心，温州　325035
第一作者单位	附属第二医院; 第二临床医学院，附属第二医院、育英儿童医院; 重症医学科
通讯作者单位	附属第二医院
第一作者的第一单位	附属第二医院
推荐引用方式 GB/T 7714	吴立琴,郑锐,李飞静,等. 吉非替尼对哮喘C57BL/6小鼠气道炎症和气道重塑的影响及作用机制[J]. 中华医学杂志,2024,104(20):1860-1867.
APA	吴立琴., 郑锐., 李飞静., 王美艳., 鲁春., ... & 戴元荣. (2024). 吉非替尼对哮喘C57BL/6小鼠气道炎症和气道重塑的影响及作用机制. 中华医学杂志, 104(20), 1860-1867.
MLA	吴立琴,et al."吉非替尼对哮喘C57BL/6小鼠气道炎症和气道重塑的影响及作用机制".中华医学杂志 104.20(2024):1860-1867.