摘要
目的观察降气平哮方对哮喘急性发作模型小鼠气道炎症的影响并探讨其可能作用机制。方法将36只BALB/c小鼠随机分为正常组,模型组,地塞米松组,降气平哮方低、中、高剂量组,每组6只。除正常组外,其余各组均予卵白蛋白建立哮喘急性发作小鼠模型。正常组、模型组予蒸馏水灌胃,降气平哮方各组分别予相应剂量降气平哮方灌胃,每日1次,连续灌胃5 d。采用全体描记系统测量小鼠支气管收缩参数增强呼吸间歇(Penh)的变化,HE染色观察小鼠肺组织病理变化,ELISA法检测小鼠肺组织匀浆中白细胞介素1β(IL-1β)、白细胞介素18(IL-18)、肿瘤坏死因子α(TNF-α)表达水平,免疫组化法检测小鼠肺组织中NOD样受体热蛋白结构域相关蛋白3(NLRP3)表达水平,Western blot法检测小鼠肺组织NLRP3炎症小体激活相关蛋白κ基因结合核因(NF-κB)、NOD样受体热蛋白结构域相关蛋白3(NLRP3)、NIMA相关激酶7(NEK7)、半胱氨酸天冬氨酸蛋白酶1(Cleaved-Caspase 1)、凋亡相关斑点样蛋白(ASC)表达。结果与正常组比较,模型组小鼠Penh升高(P<0.001),气道周围炎性细胞增多,炎症评分增加(P<0.001),肺组织匀浆中IL-1β、IL-18、TNF-α表达增加(P<0.001),肺组织NF-κB、NLRP3、NEK7、Cleaved-Caspase 1、ASC蛋白表达增加(P<0.05,P<0.01,P<0.001);与模型组相比,降气平哮方各剂量组与地塞米松组小鼠Penh减少(P<0.05,P<0.001),肺组织炎性细胞减少,炎性评分降低(P<0.001);肺组织匀浆中IL-1β、IL-18、TNF-α表达降低(P<0.05,P<0.01,P<0.001);肺组织NF-κB、NLRP3、NEK7、Cleaved-Caspase 1、ASC蛋白表达减少(P<0.05,P<0.01,P<0.001)。结论降气平哮方可改善哮喘模型小鼠肺功能及气道炎症,其作用机制可能与调节NLRP3炎症小体介导的IL-1β分泌有关。
OBJECTIVE To observe the effect of Jiangqi Pingxiao Formula on airway inflammation in mice with acute asthma and explore its possible mechanism.METHODS A total of 36 BALB/c mice were randomly divided into normal group,model group,dexamethasone group,low-dose,medium-dose and high-dose groups of Jiangqi Pingxiao Formula,with 6 mice in each group.Except for the normal group,the other groups were given ovalbumin to establish the acute asthma attack mouse model.The normal group and the model group were given distilled water by gavage,and the Jiangqi Pingxiao Formula groups were given Jiangqi Pingxiao Formula by gavage at the corresponding dose,once a day,for 5 consecutive days.Whole Body Plethysmography was used to measure the changes of enhanced respiratory interval(Pehn)of bronchial contraction parameters in mice.HE staining was used to observe the pathological changes of lung tissue in mice.ELISA method was adopted to detect the expression levels of interleukin 1β(IL-1β),interleukin 18(IL-18)and tumor necrosis factorα(TNF-α)in lung tissue homogenate of mice.Immunohistochemistry method was used to detect the expression level of NOD-like receptor pyrin domain-associated protein 3(NLRP3)in lung tissue of mice.Western blot method was employed to detect the expression of NLRP3 inflammasome activation-associated proteinκgene binding nuclear factor-κB(NF-κB),NOD-like receptor pyrin domain-associated protein 3(NLRP3),NIMA-associated kinase 7(NEK7),Caspase 1(Cleaved-Caspase 1)and apoptosis-associated speck-like protein(ASC)in lung tissue of mice.RESULTS Compared with the normal group,the Penh level of mice in the model group was increased(P<0.001),and the pathological results of lung tissue showed that the number of inflammatory cells around the airway increased,the inflammatory score increased(P<0.001),the expression of IL-1β,IL-18,and TNF-αin lung tissue homogenate increased(P<0.001),and the expression of NF-κB,NLRP3,NEK7,Cleaved-Caspase 1,and ASC proteins in lung tissue increased(P<0.05,P<0.01,P<0.001).Compared with the model group,the Penh level of mice in the Jiangqi Pingxiao Formula groups and the dexamethasone group was reduced(P<0.05,P<0.001),the number of inflammatory cells in lung tissue decreased,and the inflammatory score decreased(P<0.001);the expression of IL-1β,IL-18,and TNF-αin lung tissue homogenate decreased(P<0.05,P<0.01,P<0.001);the expression of NF-κB,NLRP3,NEK7,Cleaved-Caspase 1,and ASC proteins in lung tissue decreased(P<0.05,P<0.01,P<0.001).CONCLUSION Jiangqi Pingxiao Formula can improve lung function and airway inflammation in asthma model mice,and its mechanism may be related to regulating NLRP3 inflammasome-mediated IL-1βsecretion.
作者
陈文婷
董盈妹
单祎文
袁宸
陈文君
吴嘉宝
赵霞
CHEN Wenting;DONG Yingmei;SHAN Yiwen;YUAN Chen;CHEN Wenjun;WU Jiabao;ZHAO Xia(Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029,China;Institute of Chinese Pediatrics,Jiangsu Key Laboratory of Children’s Health and Chinese Medicine,Nanjing University of Chinese Medicine,Nanjing 210023,China;Department of Pediatrics,Liyang Hospital of Chinese Medicine,Liyang 213300,China)
出处
《南京中医药大学学报》
北大核心
2025年第2期175-180,共6页
Journal of Nanjing University of Traditional Chinese Medicine
基金
江苏省中医药科技发展计划专题研究项目(ZT202201)
江苏省高等学校基础科学(自然科学)重大项目(21KJA360002)
江苏省中医药第三批领军人才项目(SLJ0320)
常州市科技局科技项目(CJ20239005)。
作者简介
第一作者:陈文婷,女,副主任中医师,E-mail:cwtlyp@163.com;通信作者:赵霞,女,1972年生,教授、博士生导师、主任中医师,国家中医药管理局高水平中医药重点学科中医儿科学学科带头人,江苏省中医流派研究院院长,中医儿科学研究所主任,儿童健康与中医药省高校重点实验室主任,江苏省一流专业中医儿科学专业负责人,国家一流课程中医儿科学课程负责人,主要从事中医药防治儿童呼吸、消化疾病及中医体质、中医标准化研究江苏省“333工程”第二层次培养对象,江苏省中医药领军人才。中华中医药学会儿科分会副主任委员,世界中医药学会联合会儿科专业委员会副会长,全国中医药高等教育学会教师发展研究会理事长,全国中医标准技术委员会委员。国家自然科学基金会评专家。主持国家自然科学基金、教育部霍英东基金等各级课题20余项,主持参研其他课题40余项。发布国家标准1项,获国家发明专利5项,院内制剂4个,转让专利1项、院内制剂2个。获各级奖励20余项次。主编、副主编、参编教材及专著60余本,发表学术论文190余篇,E-mail:zhaoxiahy@126.com。
