内质网应激激活的iRhom2/TACE系统触发DN肾纤维化及黄葵素干预作用的研究

TNF-αconverting enzyme(TACE), functionally regulated by inactive rhomboids-2(iRhom2) is the key enzyme promoting maturation and secretion of growth factors at the post-translational level and activated under the condition of endoplasmic reticulum stress(ER-stress).ER-stress up-regulated iRhom2/TACE system function may be responsible for the pro-fibrosis and pro-inflammation growth factors’ over-secretion in the background of diabetes.Flavonoids extract of Abelmoschus esculentus(FAE) improved diabetic nephropathy. These lead us to hypothesize flavonoids extract of Abelmoschus esculentus may prevent diabetic nephropathy by post-translationally blocking the release of pro-fibrosis growth factors through inhibiting ER-stress activating iRhom2/TACE system.Our research can be divided into three parts: Part one, to observe the relationship among the endoplasmic reticulum stress of renal tissues in DN model, activity of iRhom2/TACE system and kidney disease. And to explore the intervention effect of FAE. Part two, culture human renal mesangial cells and tubular epithelial cells in vitro, to observe the effect of high concentration of glucose and AGE-BSA on changes of iRhom2/TACE system under the condition of ER-stress and the intervention role of FAE,also analyze time-effect and dose-effect relationship.Part three, to explore intervention effect of flavonoids extract of Abelmoschus esculentus on renal mesangial cell and tubular epithelial cell lines.

TNFα转化酶(TACE)是一种在翻译后水平调控生长因子分泌的关键酶，可被内质网(ER)应激激活，并受2型失活菱形分子(iRhom2)蛋白的调控。众多导致肾脏炎症与纤维化的生长因子在糖尿病背景下过度表达，是DN发生发展的主要分子机制，ER应激激活的iRhom2/TACE系统可能在这一过程中起关键的促发和驱动作用。黄葵素有减轻蛋白尿，抑制炎症因子等肾保护作用。我们推测，该作用可能是通过抑制ER应激激活的iRhom2/TACE系统，在翻译后水平阻断纤维化生长因子的过度分泌实现的。研究一：观察DN模型肾组织ER应激与iRhom2/TACE系统活性、肾脏病变之间的关系及黄葵素的干预作用。二：体外细胞培养观察高糖及AGE-BSA对人肾系膜细胞和肾小管上皮细胞ER应激iRhom2/TACE系统的变化及黄葵素的干预作用，分析时效与量效关系。三：以系膜细胞系与小管上皮细胞系为模型，探索黄葵素干预位点。

TNFα转化酶(TACE)是一种在翻译后水平调控生长因子分泌的关键酶，可被内质网(ER)应激激活，并受2型失活菱形分子(iRhom2)蛋白的调控。众多导致肾脏炎症与纤维化的生长因子在糖尿病背景下过度表达，是DN发生发展的主要分子机制，ER应激激活的iRhom2/TACE系统可能在这一过程中起关键的促发和驱动作用。黄葵素有减轻蛋白尿，抑制炎症因子等肾保护作用。我们推测，该作用可能是通过抑制ER应激激活的iRhom2/TACE系统，在翻译后水平阻断纤维化生长因子的过度分泌实现的。研究一：观察DN模型肾组织ER应激与iRhom2/TACE系统活性、肾脏病变之间的关系及黄葵素的干预作用。二：体外细胞培养观察高糖及AGE-BSA对人肾系膜细胞和肾小管上皮细胞ER应激iRhom2/TACE系统的变化及黄葵素的干预作用，分析时效与量效关系。三：以系膜细胞系与小管上皮细胞系为模型，探索黄葵素干预位点。