基于microRNA调控的活血养血汤治疗脊髓缺血再灌注损伤作用机制研究

Huoxue Yangxue Decotion is a famous old Chinese medicine doctor's experience prescription. Huoxue Yangxue Decotion has been shown to be effective in the clinical treatment of spinal cord ischemia-reperfusion injury (SCII). SCII is a complex disease because its pathogenesis and development is associated with multiple factors,Cytc—Caspase-3 mediated apoptosis pathway is one of the important mechanisms.Modern research has shown,all of which are further highly regulated by miRNA regulatory network.Our early research has shown Huoxue Yangxue Decotion adjustable SCII Cytc–Caspase- 3 mediated apoptosis pathway, prompt the miRNA regulation may be Huoxue Yangxue Decotionn treat SCII important mechanism.To further investigate the pathogenesis of SCII and the precise molecular mechanisms of Huoxue Yangxue Decotion’s therapeutic activity, in present project we will set up SCII animal models, analyze the miRNA, mRNA and protein expression profiles in SCII.Moreover, we will study the effects of Huoxue Yangxue Decotion in vivo and in vitro on the expression of those specifically expressed miRNA, mRNA, protein and signaling pathways in SCII.Furthermore, Our findings in this project will provide new insight into the molecular mechanism of pathogenesis of SCII, as well as fully elucidate the mechanism of action of how Huoxue Yangxue Decotion treats SCII.

活血养血汤是名老中医经验方，治疗脊髓缺血再灌注损伤（SCII）疗效确切。SCII是多因素、多基因、多通路参与调控的复杂性疾病，其中Cytc—Caspase-3介导的细胞凋亡通路是重要机制之一。现代研究表明miRNA调控对SCII发病机制相关的多条信号转导通路有重要调节作用。课题组前期研究表明活血养血汤可调节SCII Cytc—Caspase-3介导的细胞凋亡通路，提示miRNA调控可能是活血养血汤治疗SCII的重要机制。本课题拟应用SCII动物模型，运用基因、组学技术分析SCII的Cytc—Caspase-3介导的细胞凋亡通路中主要的miRNA-mRNA-蛋白相互作用的SCII疾病网络，以进一步明确SCII发病机制；进而用活血养血汤干预，研究其对SCII Cytc—Caspase-3介导的细胞凋亡通路特异性差异表达的miRNA及其靶基因和信号通路的影响，全面阐明活血养血汤治疗SCII的作用

老中医经验方活血养血汤干预兔SCII模型，运用基因及蛋白组学相关技术筛选差异基因和蛋白，并对其进行生物信息学分析。以阻断及再通兔第3-6腰动脉的方法建立兔SCII模型并鉴定成功，筛选出的差异miRNA、mRNA及蛋白并行相关生物信息学分析，结果表明，ocu-miRNA-191可能参与调控活血养血汤改善兔SCII模型，且相关差异miRNA、mRNA和蛋白生物进程中较多体现在单一生物进程、代谢过程及生物调节方面，分布集中于细胞器和细胞膜中，主要体现分子结合及催化活性表达，亚细胞定位主要在细胞质为主，细胞通路主要体现为脂肪酸生物合成方面，完善了活血养血汤改善SCII的作用机制，为后期研究打下基础。