吉林大学学报(医学版) ›› 2021, Vol. 47 ›› Issue (3): 537-544.doi: 10.13481/j.1671-587X.20210301

• 基础研究 •    下一篇

红景天苷对哮喘模型小鼠气道炎症的抑制作用及其机制

栾雪1,延光海2,李海波1,张波1,张巍3,黄园园1()   

  1. 1.吉林大学第一医院儿科,吉林 长春 130021
    2.吉林省过敏性常见疾病免疫与靶向重点实验室,吉林 延吉 133002
    3.吉林省长春市儿童医院健康管理中心,吉林 长春 130051
  • 收稿日期:2020-10-12 出版日期:2021-05-28 发布日期:2021-05-28
  • 通讯作者: 黄园园 E-mail:yyhuang14@mails.jlu.edu.cn
  • 作者简介:栾 雪(1993-),女,吉林省松原市人,医师,医学硕士,主要从事哮喘和感染相关疾病方面的研究。
  • 基金资助:
    国家自然科学基金项目(81860729)

Effect of salidroside on airway inflammation in mice with asthma and its mechanism

Xue LUAN1,Guanghai YAN2,Haibo LI1,Bo ZHANG1,Wei ZHANG3,Yuanyuan HUANG1()   

  1. 1.Department of Pediatrics,First Hospital,Jilin University,Changchun 130021,China
    2.Key Laboratory of Immunization and Targeting of Common Allergic Diseases of Jilin Province,Yanji 133002,China
    3.Department of Child Health Management Center,Changchun Children’s Hospital,Changchun 130051,China
  • Received:2020-10-12 Online:2021-05-28 Published:2021-05-28
  • Contact: Yuanyuan HUANG E-mail:yyhuang14@mails.jlu.edu.cn

摘要: 目的

探讨红景天苷(SAL)对小鼠急性哮喘气道炎症的干预作用,并阐明其作用机制。

方法

将50只清洁级 BALB/c 雌性小鼠随机分为对照组、卵清蛋白(OVA)组、低剂量SAL组、高剂量SAL组和地塞米松组,每组10只。于实验第1、7和14天,腹腔注射混合液200 μL(由生理盐水、10 mg OVA和1 mg氢氧化铝组成)建立小鼠哮喘模型,第21天时,将致敏小鼠置于实验玻璃箱内,以0.1 g OVA+10 mL生理盐水雾化形式激发30 min,每日1次,共7 d。低和高剂量SAL组小鼠在每次激发免疫前1 h分别腹腔注射30和60 mg·kg-1 SAL治疗液,地塞米松组小鼠腹腔注射2 mg·kg-1地塞米松治疗液,而对照组小鼠以同剂量生理盐水代替。检测各组小鼠增强呼吸间歇(Penh)值并评估小鼠气道反应性,HE染色法观察各组小鼠肺组织形态表现,直接计数法计算各组小鼠肺泡灌洗液(BALF)中中性粒细胞、嗜酸性粒细胞和淋巴细胞数,ELISA法检测各组小鼠BALF中白细胞介素1β(IL-1β)、白细胞介素4(IL-4)、白细胞介素13(IL-13)、白细胞介素17A(IL-17A)、白细胞介素18(IL-18)和白细胞介素33(IL-33)水平,Western blotting法测定各组小鼠肺组织中NOD样受体家族蛋白3(NLPR3)、凋亡相关微粒蛋白(ASC)、Caspase-1、白细胞介素1β前体(pro-IL-1β)和IL-1β蛋白表达水平。

结果

与对照组比较,OVA组小鼠Penh值明显升高(P<0.05);与OVA组比较,高剂量SAL组小鼠Penh值升高(P<0.05)。与对照组比较,OVA组小鼠气道平滑肌增厚并可见大量炎性细胞浸润;与OVA组比较,高剂量SAL组小鼠气道周围炎性细胞数减少(P<0.05)。与对照组比较,OVA组小鼠BALF中中性粒细胞、嗜酸性粒细胞和淋巴细胞数增加(P<0.05),IL-1β、IL-4、IL-13、IL-17A、IL-18和IL-33表达水平明显升高(P<0.05);与OVA组比较,高剂量SAL组小鼠BALF中中性粒细胞、嗜酸性粒细胞以和淋巴细胞数减少(P<0.05),IL-1β、IL-4、IL-13、IL-17A、IL-18和IL-33水平降低(P<0.05)。与对照组比较,OVA组小鼠肺组织中NLPR3、ASC、Caspase-1、pro-IL-1β和IL-1β蛋白表达水平明显升高(P<0.05);与OVA组比较,高剂量SAL组小鼠肺组织中NLPR3、ASC、Caspase-1、pro-IL-1β和IL-1β蛋白表达水平降低(P<0.05)。

结论

SAL可以减轻急性哮喘小鼠气道炎症,其机制可能与下调NLRP3炎性小体的表达有关。

关键词: 红景天苷, 哮喘, 气道炎症, NOD样受体家族蛋白3, 凋亡相关微粒蛋白

Abstract: Objective

To explore the interventional effect of salidroside (SAL) on acute asthmatic airway inflammation in the mice, and to clarity its mechanism.

Methods

Fifty clean BALB/c female mice were randomly divided into control group, ovalbumin (OVA) group, low dose of SAL group, high dose of SAL group and dexamethasone group, and there were 10 mice in each group. The mouse asthma model was established by intraperitoneal injection of 200 μL mixed solution (containing normal saline, 10 mg OVA, and 1 mg aluminum hydroxide) on the 1st, 7th and 14th days. On the 21st day, the sensitized mice were placed in the experimental glass box. In the box,the mice were stimulated with 0.1 g OVA+10 mL physiological saline pulverization for 30 min, once a day, lasted for 7 d.The mice in low and high doses of SAL groups were intraperitoneally injected with 30-60 mg·kg-1 SAL treatment solution 1 h before each challenge, and the mice in dexamethasone group were intraperitoneally injected with 2 mg·kg-1 dexamethasone treatment solution, while the mice in control group were replaced with the same dose of saline. The enhanced pause(Penh) values of the mice in various groups were detected and the airway reactivities of the mice were evaluated; the morphology of lung tissue of the mice in various groups was observed by HE staining; the numbers of neutrophils, eosinophils and lymphocytes in alveolar lavage fluid (BALF) of the mice in various groups were calculated by direct counting method; ELISA method was used to detect the levels of interleukin 1β (IL-1β), interleukin 4 (IL-4), interleukin 13 (IL-13),interleukin 17A (IL-17A), interleukin 18 (IL-18) ,and interleukin 33 (IL-33) in BALF of the mice in various groups; the expression level of NOD-like receptor family protein 3 (NLPR3),apoptosis-associated speck-like protein containing a CARD (ASC),Caspase-1, interleukin-1β precursor (pro-IL-1β) and IL-1β proteins in lung tissue of the mice in various groups were determined by Western blotting method.

Results

Compared with control group, the Penh value of the mice in OVA group was increased (P<0.05); compared with OVA group, the Penh value of the mice in high dose of SAL group was increased (P<0.05). Compared with control group, the airway smooth muscle of the mice in OVA group was thickened and a large amount of inflammatory cell infiltration was seen; compared with OVA group,the number of inflammatory cells around the airway of the mice in high dose of SAL group was decreased (P<0.05). Compared with control group, the numbers of neutrophils, eosinophils and lymphocytes in BALF of the mice in OVA group were increased (P<0.05).The levels of IL-1β, IL-4, IL-13, IL-17A, IL- 18, and IL-33 in BALF of the mice in OVA group were significantly increased (P<0.05);compared with OVA group, the numbers of neutrophils, eosinophils and lymphocytes in BALF of the mice in high dose of SAL group were decreased (P<0.05), the levels IL -1β, IL-4, IL-13, IL-17A, IL-18 ,and IL-33 in BALF of the mice in OVA group were decreased (P<0.05). Compared with control group, the expression levels of NLPR3, ASC, Caspase-1, pro-IL-1β ,and IL-1β proteins in lung tissue of the mice in OVA group were significantly increased (P<0.05); compared with OVA group, the expression levels of NLPR3, ASC, Caspase-1,pro-IL-1β ,and IL-1β proteins in lung tissue of the mice in high dose of SAL group were decreased (P<0.05).

Conclusion

SAL can reduce the airway inflammation in the mice with acute asthma, and its mechanism may be related to down-regulation the expression of NLRP3 inflammasome.

Key words: salidroside, asthma, airway inflammation, Nod-like receptor family pyrin domain-containing protein 3, apoptosis associated speck like protein containing a CARD

中图分类号: 

  • R562.25