[1]李程程,武怡.1,25-二羟维生素D3对哮喘小鼠Ⅱ型固有淋巴细胞的影响[J].第三军医大学学报,2019,41(12):1095-1101.
 LI Chengcheng,WU Yi.Effect of 1,25-dihydroxy vitamin D3 on expression of pro-inflammatory cytokines in type Ⅱ innate lymphoid cells in asthmatic mice[J].J Third Mil Med Univ,2019,41(12):1095-1101.
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
41卷
期数:
2019年第12期
页码:
1095-1101
栏目:
基础医学
出版日期:
2019-06-30

文章信息/Info

Title:
Effect of 1,25-dihydroxy vitamin D3 on expression of pro-inflammatory cytokines in type Ⅱ innate lymphoid cells in asthmatic mice
作者:
李程程武怡  
徐州医科大学附属医院儿科
Author(s):
LI Chengcheng WU Yi
Department of Pediatrics, Affiliated Hospital of Xuzhou Medical University, Xuzhou, Jiangsu Province, 221000, China
 
关键词:
哮喘125-二羟维生素D3Ⅱ型固有淋巴细胞白细胞介素13白细胞介素33抑瘤因子2
Keywords:
asthma 1 25-dihydroxy vitamin D3 type Ⅱ innate lymphoid cells interleukin-13 interleukin-33 suppressor of tumorigenicity 2
分类号:
R331.144; R562.25; R977.24
文献标志码:
A
摘要:

目的 观察1,25-二羟维生素D3[1,25-(OH)2-D3]对哮喘小鼠肺来源Ⅱ型固有淋巴细胞(typeⅡinnate lymphoid cells,ILC2s)及相关细胞因子的影响。方法 将24 只4~6周龄BALB/c雌性小鼠按随机数字法分为对照组、哮喘组、干预组,每组8只。哮喘组、干预组通过腹腔注射卵清蛋白致敏及雾化激发建立哮喘模型,对照组以PBS替代。干预组在激发前30 min腹腔注射1,25-(OH)2-D3混合液0.08 mL[含1,25-(OH)2-D3 0.08 μg],对照组、哮喘组以PBS替代。末次激发24 h后,行肺泡灌洗术,留取肺泡灌洗液(BALF)进行炎性细胞总数及嗜酸性粒细胞(EOS)计数;采用ELISA检测BALF中炎性因子白细胞介素33(IL-33)、白细胞介素5(IL-5)、白细胞介素13(IL-13)的表达;HE染色观察小鼠肺部病理变化;qRT-PCR检测肺组织抑瘤因子2(suppressor of tumorigenicity 2,ST-2 )、可诱导共刺激分子(inducible costimulator,ICOS)、 IL-33、 IL-5、 IL-13 mRNA表达;流式细胞仪检测肺来源ILC2s比例变化;分析BALF中EOS计数、IL-33表达与ILC2s,IL-33与IL-13的相关性。结果 哮喘组气道形态改变及炎症细胞浸润较对照组加重;干预组较哮喘组减轻。哮喘组及干预组BALF中炎性细胞总数、EOS计数、IL-33、IL-13、IL-5表达以及肺来源ILC2s(lineage-ICOS+ST-2+)比例均高于对照组(P<0.01);干预组BALF中IL-5表达与哮喘组比较差异无统计学意义,其余指标均低于哮喘组(P<0.01)。哮喘组及干预组肺组织ST-2、ICOS、IL-33、IL-5、IL-13 mRNA表达均高于对照组(P<0.01);干预组IL-5 mRNA表达高于哮喘组(P<0.05),其余指标均低于哮喘组(P<0.05)。BALF中EOS计数与ILC2s比例、IL-33表达与ILC2s比例、IL-33与IL-13表达均呈正相关关系(r=0.854、0.757、0.886,P<0.05)。结论 1,25-(OH)2-D3可通过抑制IL-33/ST-2通路调节ILC2s活化,减少IL-13分泌,改善哮喘小鼠气道炎症,而对IL-5无明显抑制。

Abstract:

Objective To investigate the effect of 1,25-dihydroxy vitamin D3 [1,25-(OH)2-D3] on the expression of pro-inflammatory cytokines in type Ⅱ innate lymphoid cells (ILC2s) in asthmatic mice. MethodsTwenty-four female BALB/c mice aged 4 to 6 weeks were randomly divided into control group, asthma group and intervention group (n=8). In the asthma group and intervention group, mouse models of asthma were established by intraperitoneal injection and aerosol inhalation of ovalbumin; PBS was used instead in the control group. In the intervention group, the mice received intraperitoneal injection of a mixture of 0.08 mL containing 0.08 μg 1,25-(OH)2-D3 30 min before ovalbumin challenge. At 24 h after the last challenge, alveolar lavage was performed, and bronchoalveolar lavage fluid (BALF) was collected for assessing the counts of the total inflammatory cells and eosinophils (EOS) and the levels of interleukin-33 (IL-33), IL-5 and IL-13 using ELISA. The pathological changes of the lungs were examined using HE staining. Quantitative real-time PCR (qRT-PCR) was used to detect the mRNA expression of suppressor of tumorigenicity 2 (ST-2), inducible costimulator (ICOS), IL-33, IL-5 and IL-13 in the lung tissues. Flow cytometry was performed to analyze the changes in the proportion of lung-derived ILC2s. The correlations of ILC2s with EOS count and IL-33 expression and the correlation between IL-33 and IL-13 in the BALF were analyzed. ResultsThe asthmatic mice showed obvious morphological changes and inflammatory cell infiltration in the airway, and these pathological changes were significantly lessened in the intervention group. The total inflammatory cell count, EOS count, levels of IL-33, IL-13, and IL-5, and the ratio of lung-derived ILC2s (lineage- ICOS+ST-2+) were significantly higher in the asthmatic group and the intervention group than in the control group (P<0.01), and were significantly lower in the intervention group than in the asthma group (P<0.01) with the exception of IL-5 level that was comparable between the 2 groups. The expression levels of ST-2, ICOS, IL-33, IL-5 and IL-13 mRNAs in the lung tissue were significantly higher in the asthma group and intervention group than in the control group (P<0.01); compared with those in the asthma group, the expression of IL-5 mRNA was significantly increased (P<0.05) and all the other indexes were significantly lowered in the intervention group (P<0.05). Positive correlations were found between EOS count and ILC2s ratio in the BALF, between IL-33 expression and ILC2s ratio, and between IL-33 and IL-13 expression in the BALF (r=0.854, 0.757, and 0.886, respectively; P<0.05). Conclusion1,25-(OH)2-D3 can inhibit the activation of ILC2s and IL-13 production by inhibiting IL-33/ST-2 pathway to alleviate airway inflammation in asthmatic mice, but it does not significantly affect IL-5 production by ILC2s.

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更新日期/Last Update: 2019-06-22