基于自噬及其转录后调控探讨补阳还五汤干预血管衰老的机制

伴随分子和细胞功能紊乱的血管衰老被视为心血管疾病(CVD)特定的危险因素，对其干预可减少或延缓严重CVD发生，具重要意义。改善气虚血瘀症是中医干预衰老的机制之一。补阳还五汤为益气活血经典代表方，其干预血管衰老的作用及机制远未阐明。我们前期研究表明补阳还五汤通过mTOR相关的自噬通路抗动脉粥样硬化形成(获国家自然基金资助)；预实验发现：衰老与年轻动脉间存在15个表达差异的miRNA；生物信息学分析显示自噬相关基因是多个差异miRNA的靶基因，其中对miR-34a研究发现可抑制自噬靶基因荧光酶活性，可诱导血管细胞衰老、调节mTOR表达。本项目将通过临床观察明确血管衰老与气虚血瘀中医症候的相关性；通过补阳还五汤干预的体内体外实验，应用二代测序、qRT-PCR及双荧光素酶报告等技术，解析补阳还五汤干预血管衰老的作用及其自噬相关转录后调控机制，为血管衰老中医防治提供新思路，为补阳还五汤作用提供新靶点

Arterial aging that accompanies by molecular and cellular disorder has been construed as specific risk factors for the cardiovascular diseases (CVD) , and thus might be targets of interventions designed to decrease or delay the occurrence of severe CVD. Qi deficiency and blood stasis are one of the traditional Chinese medicine (TCM) syndrome related to aging. As a classical Chinese medicine of tonifying Qi and activating blood, the role and mechanism of buyang huanwu decoction (BYHWD) on arterial aging, however, remains unknown. Our previous study considered that BYHWD protected against atherosclerosis formation through mTOR related autophagy pathway (supported by NSFC). Arteries miR microarray in our preliminary studies showed that a miRNAs disequilibrium started at mid-life and continued to be changed in later life and 15 miRNAs were differentially expressed in old Rhesus aortae. Furthermore, bioinformatics analysis found the predicted interaction between miRNAs and 3’ UTR of autophagy-associated gene. Among them, miR-34a suppressed the luciferase activity of the autophagy-associated gene compared with the negative control, and facilitated vascular smooth muscle cell senescence and regulated the expression of autophagy-related key factor mTOR. Thus, we propose that it might be one of the important mechanism of BYHWD on arterial aging to regulate the expression of miRNAs, consequently adjusting its target genes, autophagy signal transduction pathway. In this study, we will analyze the relationship between arterial aging and the syndrome of Qi deficiency and blood stasis in clinical research firstly. And then high throughput sequencing of miRNAs, qRT-PCR and dual luciferase report will be performance following intragastrically administering BYHWD to FXBN rat and treating BYHWD to vascular endothelial cells and vascular smooth muscle cells, in order to reveal more clearly the effect of BYHWD on arterial aging and the mechanism of autophagy associated transcriptional regulation. All of these are for exploring a new way of TCM's role on arterial aging and a new target of BYHWD.