表面活性蛋白-A在脓毒症诱导的肾小管上皮细胞损伤中的作用及机制研究

项目名称： 表面活性蛋白-A在脓毒症诱导的肾小管上皮细胞损伤中的作用及机制研究

项目编号： No.81201457

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 医学四处

项目作者： 刘娇

作者单位： 武汉大学

项目金额： 23万元

中文摘要： 脓毒症常致急性肾损伤（AKI），肾小管上皮细胞凋亡是其核心环节。研究证实表面活性蛋白-A(SP-A)能抑制脓毒症时肺上皮细胞凋亡。脂多糖（LPS）是脓毒症AKI关键致病因素，申请人发现LPS刺激后肾小管上皮细胞表达的SP-A明显升高，但其在AKI时意义尚不清楚。本项目旨在通过脓毒症AKI模型证实SP-A基因缺失对肾小管病理改变、肾小管Evans Blue渗漏、肾脏细胞凋亡等影响；通过肾小管上皮细胞体外模型，应用LIVE/DEAD染色、光谱光度及分子生物学验证重组腺病毒感染致SP-A表达改变、SP-A蛋白及不同功能片段对S-LPS刺激后细胞活性、S-LPS细胞毒性及Bax表达影响；探讨SP-A是否通过抑制TLR4介导的TRAF2-ASK1-JNK信号通路活化抑制S-LPS诱导肾小管上皮细胞凋亡。本课题揭示SP-A 在脓毒症致肾小管上皮细胞损伤中的作用和机制,为寻求防治AKI提供新的理论依据。

中文关键词： 脓毒症；急性肾损伤；表面活性蛋白－A；脂多糖；核因子－KB

英文摘要： Sepsis is the major cause of Acute Kidney Injury(AKI),Renal tubular epithelial cells apoptosis play a key role in occurrence and development of AKI.Surfactant Protein A(SP-A) was confirmed to inhibit sepsis-induced lung epithelial cells apotosis.Lipopolysaccharide（LPS） is a key pathogenic factor in sepsis-induced AKI.It was proved that renal tubular epithelial cells could express SP-A,which was increased in time and dose dependent manners induced by LPS.However,the role of SP-A in AKI still need to be explored.The project aim to explore the alteration of SP-A gene knockout mice in renal tubular pathologic detection,evans blue dye leakage and renal cells apoptosis in septic AKI models;The LIVE/DEAD staining,spectrophotometric and molecular biology methods will be used to detect the effect of SP-A and different protein fragment on the activity,smooth LPS(S-LPS) cytotoxicity and Bax expression of renal tubular epithelial cells after stimulation by S-LPS;Also to prove that whether by inhibiting TLR4 mediate TRAF2-ASK1-JNK signal pathway that SP-A inhibit renal tubular epithelial cell apoptosis.Through revealing the role and mechanism of SP-A in sepsis-induced renal tubular epithelial cell injury, the subject provides a new way to prevent AKI.

英文关键词： sepsis；acute kidney injury；surfactant protein A；LPS；NF－kB