p66Shc蛋白磷酸化调控肾小管上皮细胞凋亡通路的原位荧光分析

Protein post-translational modification (PTM) and its regulation on related signaling pathways play an important role in normal life activities, and are closely related to the occurrence and development of many diseases. Therefore, the development of efficient techniques for detecting PTM regulation on related signaling pathways is a significant cornerstone for exploring the nature of life phenomena and the molecular mechanism of disease, and is also one of the core contents of modern analytical chemistry. In this project, the molecular cause of diabetic kidney disease (DKD), p66Shc protein phosphorylation, is chosen as the research topics, typical biological macromolecules such as protein kinase, cytochrome C, and downstream ROS are chosen as the research targets. The study will focus on the phosphorylation of p66Shc protein catalyzed by protein kinase and its regulatory mechanism on ROS-related apoptosis signaling pathway in renal tubular epithelial cells under the stimulation of hyperglycemia, through the design of new signal conversion strategies and the construction of chemical / biological fluorescent probes. This will realize in situ kinetic fluorescence analysis of p66Shc protein phosphorylation-regulated apoptosis pathway in renal tubular epithelial cells, overcoming the limitations of traditional in vitro analysis which could not guarantee the original performance and the most real life state of biological samples. The implementation of this project will provide new tools to explain the signaling pathway mechanism of DKD disease, and also provide new ideas for in situ fluorescence imaging analysis of other PTM and their related signal pathways.

蛋白质翻译后修饰（PTM）及其对信号通路的调控为正常生命活动发挥着重要作用，也与多种疾病的发生、发展密切相关。针对PTM及相关信号通路调控过程发展高效的检测技术，是探究生命本质、疾病机制的重要基石，也是当前分析化学研究核心内容之一。本项目以糖尿病肾脏疾病（DKD）分子成因--p66Shc蛋白磷酸化研究为切入点，以该过程中蛋白激酶、细胞色素C等生物大分子和下游ROS为研究靶点，重点开展高血糖刺激下蛋白激酶催化p66Shc蛋白磷酸化及其对肾小管上皮细胞ROS相关凋亡通路的调控机制研究；构建化学/生物荧光探针，设计信号转换新策略，实现对p66Shc蛋白磷酸化调控肾小管上皮细胞凋亡通路的动力学原位荧光分析。本项目的实施，将克服传统离体分析难以保证生物样品原本性能和最真实生命状态的局限，为阐释DKD疾病产生的信号通路机制提供新的工具，也将为其它PTM及相关信号通路调控的原位荧光成像分析提供新思路。

蛋白质翻译后修饰（PTM）及其对信号通路的调控为正常生命活动发挥着重要作用，也与多种疾病的发生、发展密切相关。针对PTM及相关信号通路调控过程发展高效的检测技术，是探究生命本质、疾病机制的重要基石，也是当前分析化学研究核心内容之一。本项目以糖尿病肾脏疾病（DKD）分子成因--p66Shc蛋白磷酸化研究为切入点，以该过程中蛋白激酶、细胞色素C等生物大分子和下游ROS为研究靶点，重点开展高血糖刺激下蛋白激酶催化p66Shc蛋白磷酸化及其对肾小管上皮细胞ROS相关凋亡通路的调控机制研究；构建化学/生物荧光探针，设计信号转换新策略，实现对p66Shc蛋白磷酸化调控肾小管上皮细胞凋亡通路的动力学原位荧光分析。本项目的实施，克服传统离体分析难以保证生物样品原本性能和最真实生命状态的局限，为阐释DKD疾病产生的信号通路机制提供新的工具，也将为其它PTM及相关信号通路调控的原位荧光成像分析提供新思路。