维生素D对脂多糖诱导的小鼠急性肺损伤的影响

李茜; 赵燕; 王导新

doi:10.16016/j.1000-5404.201901235

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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

基础医学 | 更新时间：2026-01-09

- 维生素D对脂多糖诱导的小鼠急性肺损伤的影响
- Vitamin D alleviates lipopolysaccharide-induced acute lung injury in mice
- 第三军医大学学报 2019年41卷第14期页码：1314-1320
- 作者机构：
  
  400010 重庆，重庆医科大学附属第二医院呼吸与危重症医学科
- 作者简介：
  
  [王导新，E-mail:wangdaoxin1@163.comWANG Daoxin, E-mail:wangdaoxin1@163.com
- 基金信息：
  
  国家自然科学基金面上项目(81670071)
- DOI：10.16016/j.1000-5404.201901235
  中图分类号： R563.05;R976;977.24
- 收稿：2019-01-20，
  
  修回：2019-03-15，
  
  纸质出版：2019-07-30
- 稿件说明：
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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.

Xi LI, Yan ZHAO, Daoxin WANG. Vitamin D alleviates lipopolysaccharide-induced acute lung injury in mice[J]. Journal of Third Military Medical University, 2019, 41(14): 1314-1320.
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. DOI： 10.16016/j.1000-5404.201901235.

Xi LI, Yan ZHAO, Daoxin WANG. Vitamin D alleviates lipopolysaccharide-induced acute lung injury in mice[J]. Journal of Third Military Medical University, 2019, 41(14): 1314-1320. DOI： 10.16016/j.1000-5404.201901235.

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(

=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D

；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(

＜0.05)。相较于LPS组，VD+LPS组BALF中IL-6、TNF-α及IL-17A水平降低(

＜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(

＜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)

vitamin D

(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 L

PS 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 (

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 (

0.05).

Conclusion

Vitamin D can alleviate LPS-induced ALI in mice possibly by inhibiting pulmonary inflammatory responses mediated by Th17 cells.

关键词

Keywords

references

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