消退素D1对脂多糖诱导的小鼠肺紧密连接蛋白破坏的作用及机制

发布时间：2018-05-03 01:15

  本文选题：ALI/ARDS + 消退素D1　； 参考：《华中科技大学》2013年硕士论文

【摘要】：目的：本研究的目的是探讨消退素D1对LPS引起的急性肺损伤小鼠肺部紧密连接蛋白破坏的保护作用及机制。 方法：以小鼠气管滴注脂多糖（3mg/kg）诱导急性肺损伤模型，采用随机数字表法，将SPF级雄性小鼠（8-10w）随机分为6组（n=10）：（1）空白对照组(Control组),（2）LPS组；（3）LPS+RvD1组,（4）RvD1组,（5）LPS+RvD1+Znpp组，（6）LPS+Znpp组。消退素D1生理盐水稀释后5μg/kg，于LPS吸入前30min尾静脉注射，Znpp溶于二甲基亚砜50mg/kg于消退素30min腹腔注射，其他组给予等体积0.9%无菌生理盐水，LPS3mg/kg经切开的气管吸入后观察24h，深麻醉下放血处死小鼠，单肺灌洗，取肺组织。石蜡切片及HE染色制作肺组织病理切片观察；55℃干烤5d，检测湿干重比(w/d比)；紧密连接蛋白ZO-1和Occludin蛋白及mRNA表达水平；HO-1mRNA及蛋白表达水平；肺组织细胞凋亡率(TUNEL)；SABC免疫荧光法观察肺组织细胞间的紧密连接。 结果：病理切片显示，LPS+RvD1组肺组织损伤明显比LPS组减轻。LPS+RvD1组的肺湿干比明显小于同时间LPS组（P＜0.05）；与正常组比，LPS组紧密连接蛋白ZO-1和Occludin蛋白及mRNA表达水平明显降低（P＜0.01，P＜0.05），TUNEL示肺组织细胞凋亡率明显增加，，HO-1mRNA及蛋白表达水平稍有增加具有统计学差异(P＜0.05)；与LPS组比较，LPS+RvD1组上述指标中肺组织紧密连接蛋白ZO-1和Occludin蛋白及mRNA表达水平均升高（P＜0.05），HO-1mRNA及蛋白表达水平升高明显(P＜0.01)，肺组织细胞凋亡率明显降低；LPS+RvD1+Znpp组与LPS组比较仅有HO-1蛋白水平和mRNA表达明显下降(P＜0.01)，其他指标无统计学意义(P＞0.05)，与LPS+RvD1组比较，HO-1蛋白水平和mRNA表达下降，ZO-1和Occludin蛋白及mRNA表达水平降低(P＜0.05)，细胞凋亡增加；另外，Control组与RvD1组无统计学差异，LPS组与LPS+Znpp组无统计学差异，但LPS+Znpp组HO-1的蛋白和mRNA表达水平明显降低(P＜0.01)。免疫荧光显示LPS+RvD1组紧密连接蛋白表达量明显多于LPS组，细胞屏障破坏明显减少。 结论：消退素D1可以降低肺湿干比，减轻肺病理变化，减轻肺水肿；消退素D1预处理可以上调HO-1表达，减少紧密连接蛋白破坏，减少肺组织细胞凋亡，从而减轻急性肺损伤引起的肺水肿。
[Abstract]:Aim: to investigate the protective effect and mechanism of vanishing hormone D1 on lung tight junction protein damage induced by LPS in mice. Methods: acute lung injury model was induced by endotracheal instillation of lipopolysaccharide (3 mg / kg) in mice. SPF grade male mice were randomly divided into 6 groups for 8 to 10 weeks) (control group, lipopolysaccharide group, lipopolysaccharide D1 group, lipopolysaccharide group, RvD1 Znpp group, and control group. 5 渭 g / kg of attenuated D1 saline was injected intraperitoneally with 30min dissolved in 50mg/kg of dimethyl sulfoxide (50mg/kg) and intraperitoneally injected with dimethylsulfoxide (50mg/kg) before LPS inhalation. The other groups were treated with 0.9% aseptic saline (LPS 3 mg / kg) for 24 hours after tracheotomy. The mice were sacrificed under deep anesthesia. The mice were perfused with a single lung and the lung tissues were taken out. Paraffin sections and HE staining were used to make pathological sections of lung tissues for 5 days at 55 鈩

本文编号：1836314