芥子碱通过抑制Notch2/Notch3-Hes1信号通路改善哮喘小鼠肺组织炎症和黏液高分泌  

Sinapine alleviates lung tissue inflammation and mucus hypersecretion in asthmatic mice by inhibiting Notch2/Notch3-Hes1 signal pathway

作　　者：唐红梅 王孝芸 王剑 张沄 王志彬 袁谢芳 王星 徐国锋 覃纲 李月蛟 Tang Hongmei;Wang Xiaoyun;Wang Jian;Zhang Yun;Wang Zhibin;Yuan Xiefang;Wang Xing;Xu Guofeng;Qin Gang;Li Yuejiao(Inflammation and Allergic Diseases Research Unit,The Affiliated Hospital of Southwest Medical University,Luzhou 646000;Health Management Center,The Affiliated Hospital of Southwest Medical University,Luzhou 646000;Dept of Respiratory and Critical Care Medicine,The Affiliated Hospital of Southwest Medical University,Luzhou 646000;Dept of Otolaryngology-Head and Neck Surgery,The Affiliated Hospital of Southwest Medical University,Luzhou 646000)

机构地区：[1]西南医科大学附属医院炎症与变态反应实验室,泸州646000 [2]西南医科大学附属医院健康管理中心,泸州646000 [3]西南医科大学附属医院呼吸与危重症医学科,泸州646000 [4]西南医科大学附属医院耳鼻喉头颈外科,泸州646000

出　　处：《安徽医科大学学报》2025年第2期286-292,共7页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:82100021);四川省科技厅自然科学基金项目(编号:2022NSFSC1306);四川省科技厅重点研发计划项目(编号:2022YFS0629);西南医科大学校级科研项目(编号:2023QN043)。

摘　　要：目的探讨芥子碱对哮喘小鼠肺组织炎症和黏液分泌的影响。方法8周龄的雌性C57BL/6J小鼠随机分为对照(Control)组、卵清蛋白(OVA)组、芥子碱(Sinapine)组和Sinapine+OVA组。采用OVA+氢氧化铝[Al(OH)3]混悬液腹腔注射致敏和OVA滴鼻激发制备哮喘小鼠模型;滴鼻前1 h对Sinapine+OVA组和Sinapine组小鼠腹腔注射Sinapine溶液,OVA组和Control组小鼠给予等量0.9%氯化钠溶液。最后一次OVA滴鼻激发24 h后,HE染色观察肺组织炎症情况;PAS染色评估肺组织黏液分泌情况;荧光定量PCR(RT-qPCR)检测肺组织白细胞介素(IL)-4、IL-5、IL-13、肿瘤坏死因子-α(TNF-α)、黏蛋白5ac(Muc5ac)以及Notch通路关键基因缺口受体(Notch)1、Notch2、Notch3和hes家族bHLH转录因子1(Hes1)mRNA表达水平;Western blot法检测Notch1、Notch2、Notch3和Hes1蛋白表达水平。结果与Control组比较,OVA组小鼠肺组织炎症评分和PAS评分升高(P<0.001),IL-4、IL-5、IL-13、TNF-α和Muc5ac mRNA表达水平增加(P<0.05),Notch通路中Notch2、Notch3和Hes1的mRNA和蛋白表达水平升高(P<0.001),而Notch1 mRNA和蛋白表达水平差异无统计学意义;与OVA组比较,Sinapine+OVA组小鼠肺组织炎症评分和PAS评分降低(P<0.001),IL-4、IL-5、IL-13、TNF-α和Muc5ac mRNA表达下降(P<0.05),Notch通路中Notch2、Notch3和Hes1的mRNA和蛋白表达降低(P<0.05),而Notch1 mRNA和蛋白表达水平差异无统计学意义。结论芥子碱可改善哮喘小鼠肺组织炎症和黏液高分泌,其作用机制可能与抑制Notch2/Notch3-Hes1信号通路有关。Objective To investigate the effects of sinapine on lung tissue inflammation and mucus hypersecretion in asthmatic mice.Methods Eight-week-old female C57BL/6J mice were randomly divided into Control group,ovalbumin(OVA)group,Sinapine group,and Sinapine+OVA group.The asthmatic mice model were established by intraperitoneal injection of OVA combined with aluminum hydroxide[Al(OH)3]suspension and OVA nasal stimulation.One hour before OVA nasal stimulation,the mice in Sinapine+OVA group and Sinapine group were intraperitoneally injected with sinapine solution,and the mice in OVA group and Control group were treated with the same dose of 0.9%sodium chloride solution.24 hours after the last OVA stimulation,the inflammation of lung tissue of mice were observed by HE staining;the mucus secretion were evaluated by PAS staining;the mRNA expression levels of Interleukin-4(IL-4),Interleukin-5(IL-5),Interleukin-13(IL-13),tumor necrosis factor-alpha(TNF-α),Mucin 5ac(Muc5ac),and the mRNA of the key genes of Notch pathway such as Notch receptor 1(Notch1),Notch receptor 2(Notch2),Notch receptor 3(Notch3),and hes family bHLH transcription factor 1(Hes1)in lung tissues were detected by real-time fluorescent quantitative PCR(RT-qPCR);the expression levels of Notch1,Notch2,Notch3 and Hes1 proteins were determined by Western blot.Results Compared with Control group,the inflammation score and PAS score of lung tissues of mice in OVA group increased(P<0.001);the mRNA expression levels of IL-4,IL-5,IL-13,TNF-α,and Muc5ac of mice in OVA group were enhanced(P<0.05);the mRNA and protein expression levels of Notch2,Notch3,and Hes1 of mice in OVA group significantly increased(P<0.001),while there was no significant difference in the mRNA and protein expression levels of Notch1.Compared with OVA group,the inflammation score and PAS score of lung tissues of mice in Sinapine+OVA group decreased(P<0.001);the mRNA expression levels of IL-4,IL-5,IL-13,TNF-α,and Muc5ac of mice in Sinapine+OVA group were reduced(P<0.05);the mRNA and protein expres

关 键 词：芥子碱 哮喘 Notch通路 促炎细胞因子 黏蛋白5AC 气道炎症 黏液高分泌 

分 类 号：R562.2[医药卫生—呼吸系统]