高尔基体形态结构变化与小鼠内毒素性急性肺损伤的关系

Relationship between changes in Golgi apparatus morphological structure and endotoxin-induced acute lung injury in mice

目的评价高尔基体形态结构变化与小鼠内毒素性急性肺损伤的关系。方法健康清洁级雄性C57BL/6J小鼠20只,体重18~20 g,6~8周龄,采用随机数字表法分为2组(n=10):假手术组(Sham组)和内毒素性急性肺损伤组(ALI组)。ALI组静脉注射LPS 10 mg/kg,Sham组给予等容量生理盐水0.5 ml。注射LPS后12 h时处死小鼠,取肺组织,测定ROS含量和湿重/干重(W/D)比值;采用HE染色观察小鼠肺组织病理学变化;分别采用Western blot法和qPCR法检测高尔基体基质蛋白130(GM130)、高尔基体蛋白97(Golgin97)和Ⅱ类α-甘露糖苷酶(MAN2A1)及其mRNA的表达水平;采用透射电镜观察高尔基体形态结构。结果与Sham组比较,ALI组肺组织ROS含量和W/D比值升高,GM130、MAN2A1及Golgin97及其mRNA表达下调(P<0.01),肺组织病理学损伤加重,高尔基体膜囊发生肿胀,高尔基体碎片弥散在细胞质中。结论小鼠内毒素性急性肺损伤的机制可能与高尔基体形态结构改变有关。

Objective To evaluate the relationship between changes in Golgi apparatus morphological structure and endotoxin-induced acute lung injury(ALI)in mice.Methods Twenty healthy male C57BL/6J mice,weighing 18-20 g,aged 6-8 weeks,were divided into 2 groups(n=10 each)using a random number table method:sham operation group(group Sham)and endotoxin-induced ALI group(group ALI).Lipopolysaccharide(LPS)10 mg/kg was injected intravenously in group ALI,while the equal volume of normal saline 0.5 ml was given instead in group Sham.The animals were sacrificed at 12 h after LPS injection and the lung tissues were taken for detection of the content of reactive oxygen species(ROS)and wet to dry weight ratio(W/D ratio),for observation of the pathological changes(using HE staining)and Golgi apparatus morphological structure(with a transmission electron microscope)and for determination of expression of Golgi matrix protein 130(GM130),Golgin97 and mannosidase alpha class II member 1(MAN2A1)and its mRNA(by Western blot and quantitative polymerase chain reaction).Results Compared with group Sham,ROS content and the W/D ratio in lung tissues were significantly increased,GM130,MAN2A1,Golgin97 protein and its mRNA expression were down-regulate(P<0.01),the pathological changes of lung tissues were accentuated,the Golgi cisternae was swollen,and Golgi fragments were dispersed in the cytoplasm in group ALI.Conclusion The mechanism of endotoxin-induced ALI may be related to the changes in Golgi apparatus morphological structure.