脂多糖下调过氧化物酶增殖体激活受体γ在大鼠原代肺泡Ⅱ型上皮细胞的表达  

LPS stimulation down-regulates PPARγ expression in rat type Ⅱ alveolar epithelial cells

作　　者：肖波[1] 王建春[1] 王关嵩[1] 邓朝霞[1] 徐静[1] 钱桂生[1] 

机构地区：[1]第三军医大学新桥医院呼吸内科研究所,全军呼吸病研究重点实验室,重庆400037

出　　处：《第三军医大学学报》2009年第20期1960-1963,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(30570808);全军医学科研“十一五”计划科技攻关项目(06G083)~~

摘　　要：目的观察脂多糖(lipopolysac charide,LPS)作用下大鼠肺泡Ⅱ型上皮(typeⅡalveol arepith elial,AT-Ⅱ)细胞过氧化物酶增殖体激活受体γ(PPARγ)的表达变化情况。方法通过酶消化分离和免疫黏附法纯化原代培养AT-Ⅱ细胞,并进行碱性磷酸酶、肺表面活性物质相关蛋白A(SP-A)和电镜鉴定。采用RT-PCR法检测细胞经LPS作用后PPARγ、TNF-α、SP-AmRNA的改变,Westernblot法检测LPS作用后细胞PPARγ表达,ELISA法检测TNF-α蛋白表达。结果经碱性磷酸酶、SP-A和电镜鉴定,原代培养AT-Ⅱ细胞成功;在致炎因子LPS作用下,PPARγmRNA和蛋白表达均下降,这一变化伴随炎性因子TNF-α的升高。结论在LPS作用下,AT-Ⅱ细胞内PPARγ表达降低,提示PPARγ参与炎症反应。Objective To determine the effect of LPS stimulation on the expression of PPARγ in rat type Ⅱ alveolar epithelial （AT-Ⅱ） cells. Methods Rat lung tissues were digested by trypsase and collagenase. AT-Ⅱ cells were isolated and purified by immuno-adherence, and then primarily cultured AT-Ⅱ cells were identified by alkaline phosphatase （AKP）, pulmonary surfactant-associated protein A （SP-A） staining and electron microscopy. PPARγ, TNF-α and SP-A mRNA levels in AT-Ⅱcells were assayed by RT-PCR after being activated by lipopolysaccharide （LPS） stimulation, while the PPARγ and TNF-α protein levels in culture supernatants were measured by Western blotting and ELISA respectively. Results Primary culture of rat AT-Ⅱ cells were successfully established as confirmed by the expression of AKP and SP-A and unltrastructural features. PPARγ expression in AT-Ⅱ cells were decreased when stimulated with LPS, which were accompanied by increased TNFα expression. Conclusion Pro-inflammatory substance LPS can markedly inhibit the expression of PPARγ in AT-Ⅱcells, which could be involved in the loss of control of inflammation.

关 键 词：过氧化物酶增殖体激活受体Γ 肺泡Ⅱ型上皮细胞 炎症 

分 类 号：R322.35[医药卫生—人体解剖和组织胚胎学] R345.46[医药卫生—基础医学]