基于lncRNA VPRT-AS调控SIRT1/KLF4信号探讨艳山姜抑制糖尿病血管钙化的作用机制研究

糖尿病患者血管呈典型中层钙化，血管平滑肌细胞（VSMCs）表型转化为血管钙化的关键环节。lncRNA芯片技术筛选出表达上调的lnc VPRT-AS，发现其促进VSMCs分化标志基因KLF4的表达。研究显示，SIRT1调控KLF4的基因转录，而SIRT1的苏木化修饰是其重要的转录修饰方式，预实验发现敲低VPRT-AS促进SIRT1的苏木化，抑制SIRT1/KLF4信号的转录。因此，本研究提出以VSMCs表型转化介导血管钙化为切入点，系统分析药物基于lncRNA VPRT-AS调控SIRT1/KLF4级联信号的转录机制，可能是艳山姜逆转VSMCs表型转化，抑制糖尿病血管钙化的重要分子机制假说。课题利用转基因动物，通过干预相关基因的表达，结合CHIP实验及双荧光素酶报告分析艳山姜抑制糖尿病血管钙化的作用及机制。研究为民族药物艳山姜血管药理学活性提供科学技术分析，有利于民族医药特色的创新药物探索。

Vascular calcification is an important link between cardiovascular and diabetes，vascular smooth muscle cell (VSMCs) phenotype swithing is a major factor for vascular calcification of diabetes mellitus. We found lncVPRT-AS expression level was significantly increased in calcified thoracic aorta of diabetes rats by Using lncRNA-microarray technology, and it’s particularly expression in VSMCs.The result suggested that knockdown of VPRT-AS that decreased the transcription of KLF4 which has a critical regulatory effects on VSMCs phenotype swithing.The present study has demonstrated that SUMO-1 mediated sumoylation of SIRT1 is the important post translational modifications of SIRT1, and our data has showed that VPRT-AS-knockdown regulated SIRT1 sumoylation level and SIRT1/KLF4 signaling pathway. Therefore, we propose the hypothesis that ethnicmedicine EOFAZ inhibits vascular calcification of diabetes through downregulation of lncVPRT-AS, which regulated the SIRT1 sumoylation, and inhibited the expression level of KLF4. In this study, we applied db/db mouse, utilizing CHIP and Luciferase assays to investigate the intervention mechaism of EOFAZ on vascular calcification of diabetes, which can provide the experimental reference about discovering of EOFAZ and provide theoretical reference about the therapeutical potential of ethnicmedicine on vascular calcification.