甲基莲心碱调控cGAS-STING信号通路对哮喘幼年大鼠气道炎症的影响  

Impact of neferine on airway inflammation in asthmatic young rats by regulating cGAS-STING signal pathway

作　　者：张婵[1] 祝颖 李爱丽 ZHANG Chan;ZHU Ying;LI Aili(Heze Medical College,Heze 274000,China;Department of Pediatrics,Affiliated Hospital of Heze Medical College,Heze 274000,China)

机构地区：[1]菏泽医学专科学校,274000 [2]菏泽医学专科学校附属医院儿科,274000

出　　处：《免疫学杂志》2023年第10期879-885,共7页Immunological Journal

基　　金：山东省医药卫生科技发展计划(202006011247)

摘　　要：目的分析甲基莲心碱对哮喘幼年大鼠气道炎症的影响,并分析其对cGAS-STING信号通路的作用。方法将50只SD大鼠随机分为对照组、模型组、甲基莲心碱低(20 mg/kg)、高剂量(40 mg/kg)组、甲基莲心碱高剂量+2′3′-cGAMP组(40 mg/kg甲基莲心碱+500μg/kg 2′3′-cGAMP)。除对照组外,其余各组均构建哮喘模型,按照上述条件给药,测定大鼠支气管肺泡灌洗液中CysLTs、CysLTR1水平,对大鼠肺组织进行HE染色与PAS染色以评估大鼠支气管周围、血管周围炎性浸润以及杯状细胞的生成;ELISA试剂盒测定大鼠肺组织IL-4、IFN-γ水平,Western blot法测定大鼠肺组织cGAS-STING通路相关蛋白水平。结果与对照组比较,模型组大鼠血管内壁与支气管可见大量炎性细胞浸润,杯状细胞增生明显;甲基莲心碱低、高剂量组大鼠气道炎症减轻;与甲基莲心碱高剂量组比较,甲基莲心碱高剂量+2′3′-cGAMP组大鼠气道炎症加重。与对照组比较,模型组大鼠肺泡灌洗液中CysLTs水平、CysLTR1水平、肺组织炎症评分、黏液评分、IL-4水平、cGAS蛋白表达、STING蛋白表达升高,肺组织IFN-γ水平降低(P<0.05);与模型组比较,甲基莲心碱低剂量组、甲基莲心碱高剂量组大鼠上述改变均被逆转,且呈剂量依赖性;与甲基莲心碱高剂量组比较,甲基莲心碱高剂量+2′3′-cGAMP组大鼠肺组织炎症评分、黏液评分、IL-4水平、cGAS、STING蛋白表达、肺泡灌洗液中CysLTs、CysLTR1水平升高,肺组织IFN-γ水平降低(P<0.05)。结论甲基莲心碱可能通过抑制cGASSTING通路活化缓解新生哮喘大鼠气道炎症。This study was performed to analyze the impact of neferine on airway inflammation in young rats with asthma,and its effect on cGAS-STING signal pathway.Total of 50 SD rats were randomly grouped into control group,model group,low-dose neferine group(20 mg/kg),high-dose neferine group(40 mg/kg),high-dose neferine+2′3′-cGAMP group(40 mg/kg neferine+500μg/kg 2′3′-cGAMP).In addition to the control group,asthma models were established in all other groups.ELISA was used to measure the levels of CysLTs and CysLTR1 in the bronchoalveolar lavage fluid of rats;HE staining and PAS staining were performed on the lung tissue of rats to evaluate the formation of goblet cells and the inflammatory infiltration around the bronchus and blood vessels;the levels of IL-4 and IFN-γ in lung tissue were measured with ELISA kits;the levels of cGAS-STING pathway related proteins in lung tissue were detected by Western blotting.Compared with the control group,a large number of inflammatory cells infiltrated into the inner wall of bronchi and blood vessels,and goblet cells proliferated obviously in the model group;the airway inflammation of rats in the two neferine groups were reduced;compared with the high-dose neferine group,the airway inflammation of rats in the high-dose neferine+2′3′-cGAMP group was aggravated.Compared with the control group,the levels of CysLTs and CysLTR1 in the alveolar lavage fluid,lung inflammation score,mucus score,IL-4 level,and the expression of cGAS and STING proteins in lung tissue of the model group were increased,while the level of IFN-γ in lung tissue was decreased(P<0.05).Neferine(both low-dose and high-dose)could reverse these changes mentioned above in model rats(P<0.05),in a dose-dependent manner.While 2′3′-cGAMP could suppress these effects of neferine(P<0.05).Taken together,neferine may alleviate airway inflammation in neonatal asthmatic rats by inhibiting the activation of cGAS-STING pathway.

关 键 词：甲基莲心碱 哮喘 气道炎症 cGAS-STING通路 

分 类 号：R285.5[医药卫生—中药学]