同型半胱氨酸经NUPR1激活TIA1调控特异性lncRNA介导足细胞自噬水平升高的机制

自噬水平升高是同型半胱氨酸(Hcy)致肾损伤的重要机制。前期证实lncRNA参与了Hcy引起足细胞损伤的调控，但机制未清。T细胞内抗原-1(TIA1)可调控细胞自噬并受多种转录因子的调控，故推测：Hcy经转录因子NUPR1激活TIA1磷酸化上调特异性lncRNA的表达是Hcy引起自噬水平升高介导足细胞损伤的重要机制。为验证该假说，微阵列技术筛选并确定lncDACT1是Hcy引起肾损伤的特异性lncRNA，构建其过表达或沉默载体转染细胞后分析自噬相关基因的变化，明确lncDACT1在Hcy引起足细胞自噬水平升高中的作用；采用以阻断为主的策略转染NUPR1和TIA1过表达和沉默载体，ChIP-seq分析其结合情况，确定NUPR1激活TIA1磷酸化上调lncDACT1引起足细胞自噬水平升高的机制，酵母双杂交筛选并结合质谱阐明ANXA7具有协同NUPR1作用，为防治Hcy致肾损伤提供依据。

Elevated autophagy is an important mechanism of renal injury induced by homocysteine (Hcy). It was previously confirmed that lncRNA was involved in the regulation of Hcy-induced podocyte injury, but the mechanism is unclear. T-cell antigen-1 (TIA1) can modulated autophagy and be regulated by many transcript factors. Therefore, it is speculated that Hcy promoted lncDACT1 expression by NUPR1-activated TIA1 phosphorylation may be an important mechanism in Hcy-induced podocyte injury. To verify this hypothesis, microarray was used to screen and confirm that lncDACT1 is the specific lncRNA of kidney injury caused by Hcy, then the changes of autophagy-related genes were analyzed after the cells were transfected with overexpression or silencing vector to clarify the role of lncDACT1 in Hcy-induced autophagy in podocytes; Taking block strategy, podocyte was transfected with recombination and RNA inference vector of NUPR1 and TIA1 to investigate the mechanisms of increasing lncDACT1 expression by NUPR1-activated phosphorylation of TIA1; Identifying the mechanism of ANXA7 regulated TIA1 phosphorylate by yeast-two-hybrid screening and mass-spectrometric technique in podocyte. Above all, we provide experimental basis for the prevention and treatment of renal injury induced by Hcy.