淫羊藿总黄酮调控MALAT1/miR-26靶向作用EphA2影响成骨细胞骨架稳态的机制研究

骨髓炎所致骨不连是临床骨科亟待解决的难题。促红细胞生成素产生肝细胞受体EphA2及RhoA/Rock信号通路对成骨细胞骨架稳态具有重要作用。申请人前期研究结果提示，脂多糖诱导可上调成骨细胞EphA2及RhoA表达，促使细胞骨架F-actin解聚，而miR-26与MALAT1相互作用可调控靶基因EphA2表达。淫羊藿总黄酮能维护细胞骨架稳态促进骨修复，但确切机制不明。本项目拟采用脂多糖诱导成骨细胞模型，应用质粒转染、双荧光素酶实验及siRNA干扰技术，研究MALAT1/miR-26调控靶基因EphA2影响细胞骨架稳态失衡机制；探讨淫羊藿总黄酮作用于MALAT1/miR-26/EphA2，调控下游RhoA/Rock信号通路，进而促进骨基质形成的机制。阐明淫羊藿总黄酮调控炎症损伤-骨架稳态-骨修复的作用机制，为“补肾健骨”中药的科学内涵阐述及应用提供新的思路和实验依据，促进骨髓炎临床研究。

Bone nonunion caused by osteomyelitis is an urgent problem in clinical orthopedics. Erythropoietin-producing hepatocellular receptor A2（EphA2）and RhoA / Rock signaling pathway play an important role in the homeostasis of cytoskeleton. Our previous research found that LPS induction could up-regulate the expression of EphA2 and RhoA in osteoblasts, then lead to the depolymerization of F-actin. Meanwhile, miR-26 and MALAT1 could interact to regulate the expression of EphA2. The total flavonoids of Epimedium could maintain the homeostasis of cytoskeleton and promote bone repair, but the exact mechanism is unknown. In this project, the osteoblast model induced by LPS, plasmid transfection, double luciferase experiment and siRNA interference technology will be used to study the mechanism of MALAT1 / miR-26 regulating target gene EphA2 affecting the homeostasis imbalance of cytoskeleton. The research will clarify the effect of total flavonoids of Epimedium on MALAT1 / miR-26 / EphA2, to regulate the downstream RhoA / Rock signal pathway, to promote the repair mechanism of bone matrix formation, to clarify the mechanism of total flavonoids of Epimedium regulating inflammation cytoskeleton bone formation. The results may provide new ideas and experimental basis for the scientific connotation and application of herb invigorating the kidney and strengthening the bone, and promote the clinical research of osteomyelitis.