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.

维生素D对脂多糖诱导的小鼠急性肺损伤的影响(/HTML )




Vitamin D alleviates lipopolysaccharide-induced acute lung injury in mice
LI Xi ZHAO Yan WANG Daoxin

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


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。

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