脂多糖对大鼠肺损伤及肺组织中AQP1和AQP5表达的影响

doi:10.13481/j.1671-587x.20160212

摘要/Abstract

摘要：

目的: 探讨大鼠经静脉注射脂多糖(LPS)对急性肺损伤(ALI)及肺组织中水通道蛋白1(AQP1)和水通道蛋白5(AQP5)表达的影响,阐明其作用机制。方法: SPF级2月龄雄性Wistar大鼠48只随机分为对照组和LPS组(n=24),分别经尾静脉注射生理盐水和LPS,每组分别于注射后2、6、12和24 h时间点采集标本,每个时间点6只。HE染色观察肺组织病理学变化;测定大鼠肺湿/干质量(W/D)比、肺渗透指数;ELISA法检测血清肿瘤坏死因子α(TNF-α)和巨噬细胞炎性蛋白1α(MIP-1α)水平;采用Western blotting、免疫组织化学和Real-Time PCR方法检测大鼠肺组织中AQP1和AQP5蛋白及mRNA表达水平。结果: 与对照组比较,LPS组大鼠血清TNF-α和MIP-1α水平在注射LPS后2、6和12h时间点均明显升高(P<0.05),至24h时逐渐恢复正常。HE染色,注射LPS后2h大鼠肺组织出现水肿和炎性细胞浸润,以12h时间点最明显;LPS组各时间点大鼠肺W/D比、肺渗透指数均较对照组明显升高(P<0.05),12h时间点最明显。与对照组比较,LPS组各时间点大鼠肺组织中AQP1和AQP5 mRNA及蛋白表达水平均明显下降(P<0.01),以12h时间点下降最明显。结论: LPS可导致大鼠ALI和肺组织中AQP1及AQP5表达水平下调,并参与了肺水肿的形成。

关键词: 脂多糖, 肺损伤, 水通道蛋白1, 水通道蛋白5

Abstract:

Objective: To investigate the effects of lipopolysaccharide (LPS) on the acute lung injury (ALI) and expressions of aquaporin 1 (AQP1) and aquaporin 5 (AQP5) in lung tissue of the rats. Methods: Forty-eight SPF grade male Wistar rats were randomly divided into control group and LPS group (n=24).The rats in LPS group were intravenously injected with LPS to induce ALI models, and the rats in control group were injected with saline.The rats were sacrificed at 2, 6, 12 and 24 h, and the samples were collected after the successful modeling.The pathological changes of lung tissue were observed with HE staining; the lung wet/dry weight (W/D) ratio and lung permeability index were detected; ELISA was used to detect the levels of TNF-α and MIP-1α.The expression levels of AQP1 and AQP5 protein and mRNA were measured by Western blotting, immunohistochemistry and Real-Time PCR methods.Results: Compared with control group, the TNF-α and MIP-1α levels in LPS group were significantly elevated at 2, 6 and 12 h (P<0.05), and at 24 h they were gradually reduced to the normal level.The HE staining results showed the alveolar and interstitial edema at 2 h after LPS injection, obviously in 12 h.The lung W/D ratios and pulmonary permeability indexes at different time points in LPS group were significantly higher than those in control group (P<0.05), and they reached the peak at 12 h.The expression levels of AQP1 and AQP5 mRNA and protein in lung tissue of the rats at different time points in LPS group were significantly lower than those in control group (P<0.01).Conclusion: LPS can induce ALI in the rats and down-regulate the expressions of AQP1 and AQP5; LPS is involved in the formation of pulmonary edema.

Key words: lipopolysaccharide, lung injury, aquaporins 1, aquaporins 5

中图分类号:

R563

陈国兵, 吴谨准, 连珠兰, 占珠琴, 白海涛. 脂多糖对大鼠肺损伤及肺组织中AQP1和AQP5表达的影响[J]. 吉林大学学报(医学版), 2016, 42(02): 250-254.

CHEN Guobing, WU Jinzhun, LIAN Zhulan, ZHAN Zhuqin, BAI Haitao. Effects of lipopolysaccharide on lung injury and expressions of AQP1 and AQP5 in lung tissue of rats[J]. Journal of Jilin University Medicine Edition, 2016, 42(02): 250-254.

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