[1]李茜,赵燕,王导新.维生素D对脂多糖诱导的小鼠急性肺损伤的影响[J].第三军医大学学报,2019,41(14):1314-1320.
 LI Xi,ZHAO Yan,WANG Daoxin.Vitamin D alleviates lipopolysaccharide-induced acute lung injury in mice[J].J Third Mil Med Univ,2019,41(14):1314-1320.
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维生素D对脂多糖诱导的小鼠急性肺损伤的影响(/HTML )
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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14期
页码:
1314-1320
栏目:
基础医学
出版日期:
2019-07-30

文章信息/Info

Title:
Vitamin D alleviates lipopolysaccharide-induced acute lung injury in mice
作者:
李茜赵燕王导新
重庆医科大学附属第二医院呼吸与危重症医学科
Author(s):
LI Xi ZHAO Yan WANG Daoxin

Department of Respiratory and Critical Care Medicine, the Second Affiliated Hospital of Chongqing Medical University, Chongqing, 400010, China

关键词:
维生素D急性呼吸窘迫综合征急性肺损伤Th17细胞
Keywords:
分类号:
R563.05;R976;977.24
文献标志码:
A
摘要:

目的 探讨维生素D(vitamin D, VD)对脂多糖(lipopolysaccharide, LPS)诱导的小鼠急性肺损伤的影响及其可能的机制。方法 将45只雄性C57BL/6小鼠按随机数字表法分为3组(n=15):对照组、LPS组、维生素D补充组(VD+LPS组)。LPS构建小鼠急性肺损伤模型前,VD+LPS组予以25 μg/kg灌胃补充3剂1,25二羟维生素D3;对照组及LPS组予以相同体积的0.1%二甲基亚砜(dimethylsulfoxide, DMSO)灌胃。气管插管给予3 mg/kg LPS构建小鼠急性肺损伤模型,造模后2 d处死小鼠。检测支气管肺泡灌洗液(bronchoalveolar lavage fluid, BALF)细胞总数、中性粒细胞比例和蛋白浓度。酶联免疫吸附实验(enzyme-linked immunosorbent assay, ELISA)检测BALF中白细胞介素-6(interleukin-6, IL-6)、肿瘤坏死因子-α(tumor necrosis factor-α, TNF-α)及白细胞介素-17A(IL-17A)水平。HE染色观察肺组织病理改变,以肺组织湿/干质量比(wet to dry ratio, W/D)评估肺组织水肿情况,流式细胞术检测肺组织辅助性T细胞17(T helper 17 cells, Th17)比例。Western blot和qPCR检测Th17细胞特征性转录因子维甲酸相关孤核受体γt(retinoic acid-related orphan receptor γt, RORγt)表达水平。结果 补充维生素D明显减轻LPS造模小鼠肺损伤严重程度。与LPS组比较,VD+LPS组BALF细胞总数、中性粒细胞比例、蛋白浓度降低(P<0.05)。相较于LPS组,VD+LPS组BALF中IL-6、TNF-α及IL-17A水平降低(P<0.05)。与LPS组相比,VD+LPS组Th17细胞比例明显降低,RORγt蛋白及核酸表达显著下降(P<0.05)。结论 维生素D可能通过抑制Th17细胞介导的炎症反应以减轻LPS诱导的小鼠急性肺损伤。

Abstract:
Objective To investigate the effect of vitamin D on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice and explore the possible mechanism. Methods Forty-five male C57BL/6 mice were randomized equally into control group, LPS group and vitamin D supplement (VD+LPS) group. In VD+LPS group, the mice were given 3 doses of 1,25(OH)2 vitamin D3 (25 μg/kg) via gavage administrartion at 1, 24 and 48 h before LPS (3 mg/kg) challenge via tracheal intubation, and those in the other 2 groups received lavage with 0.1% dimethylsulfoxide (DMSO) prior to or without LPS challenge. The mice were sacrificed in 2 d after LPS challenge for examination of the total cell count, proportion of neutrophils and protein concentration in the bronchoalveolar lavage fluid (BALF); Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and IL-17A in the BALF. HE staining was used to observe the pathological changes in the lung tissues, and lung edema was assessed by measuring the wet-to-dry ratio (W/D). Flow cytometry was used to analyze the percentage of T helper 17 (Th17) cells in the lungs. Western blotting and quantitative real-time PCR (qPCR) were performed to detect the expression of retinoic acid-related orphan receptor γt (RORγt) in the lung homogenate. Results Pretreatment of the mice with vitamin D prior to LPS stimulation significantly reduced the severity of LPS-induced lung injury. Compared with those in LPS group, the total cell count, percentage of neutrophils, protein concentration, and levels of IL-6, TNF-α and IL-17A in the BALF were all decreased in VD+LPS group (P<0.05). The percentage of Th17 cells in the lung tissue and the pulmonary expression of RORγt at both the mRNA and protein levels were significantly lower in VD+LPS group than in LPS group (P<0.05). Conclusion Vitamin D can alleviate LPS-induced ALI in mice possibly by inhibiting pulmonary inflammatory responses mediated by Th17 cells.
 

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更新日期/Last Update: 2019-07-22