基于FKBP52/GRα/miRNA-708信号通路探讨激素性股骨头坏死的分子机制及生骨再造丸的干预效应

Steroid Induced Avascular Necrosis of Femoral Head(SANFH) is a bone disease of ischemic and necrotic of femoral head due to the use of Glucocorticoid in clinical treatment irregularly. The imbalance of osteogenic and adipogenic differentiation of bone marrow mesenchymal stem cells is (BMSCs) one of the key factors in this disease.The FKBP52/GRα/miRNA-708 signaling pathway plays a key role in the osteogenic-adipogenic differentiation of BMSCs. According to the “YANG Xu-TAN Shi-XUE Yu theory” of traditional Chinese medicine and basing on the results obtained in our recent experiments, we hypothesis that molecular biological mechanism of SANFH treated by the Sheng Gu Zai Zao Pill with “TONG Yang-LI Shi-HUO Xue” function is inhibit adipogenic differentiation of BMSCs, promoting their osteogenic differentiation for therapeutic purposes by inhibiting the FKBP52/GRα/miRNA-708 signaling pathway. This research intends to investigate therelationship between FKBP52/GRα/miRNA-708 signaling pathway and SANFH, hunt for miRNA-708 target gene and downstream signaling pathway in the process of osteogenic-adipogenic differentiation of BMSCs, and illustrate action mechanism of Sheng Gu Zai Zao Pill in treating SANFH. The research will be great significance of guidance to clarify the pathogenesis of SANFH as well as give full play to the advantages of Traditional Chinese medicine treatment.

激素性股骨头坏死（SANFH）是因糖皮质激素不规范使用致股骨头缺血坏死的骨疾病。骨髓间充质干细胞（BMSCs）的成骨-成脂分化失衡是SANFH发生的关键因素之一。FKBP52/GRα/miRNA-708信号通路在BMSCs的成骨-成脂分化过程中起到关键作用。本项目以中医“阳虚-痰湿-血瘀”理论为指导，基于前期研究结果提出具有“通阳-利湿-活血”功效的生骨再造丸治疗SANFH的分子生物学内涵是“通过调控FKBP52/GRα/miRNA-708信号通路来抑制BMSCs的成脂分化，促进成骨分化以达到治疗目的”的假说。拟采用基因沉默、转染、芯片检测等技术研究FKBP52/GRα/miRNA-708信号通路与SANFH的关系，寻找BMSCs的成骨-成脂分化中miRNA-708的下游靶基因及信号通路，阐明生骨再造丸干预的机制。本项目对明确SANFH的发病机制及发挥中医药治疗的优势具有重要意义。

激素性股骨头坏死（SANFH）是糖皮质激素不规范使用导致的股骨头缺血坏死的骨疾病。骨髓间充质干细胞（BMSCs）的成骨-成脂分化失衡是SANFH发生的关键因素之一。本项目以中医“阳虚-痰湿-血瘀”理论为指导，采用基因沉默、慢病毒、高通量基因测序等技术研究FKBP52/GRα/miRNA-708信号通路与SANFH的关系，寻找BMSCs的成骨-成脂分化中miRNA-708的相关基因及信号通路，并应用具有“通阳-利湿-活血”功效的生骨再造丸进行干预。结果表明LPS联合甲泼尼龙琥珀酸钠法可以成功复制SANFH模型；全骨髓培养法可以成功提取BMSCs；基因测序获得 NPY、TMEM164、SLC7A11、RAB27B、COLEC12、GPRC5B、ECRG4、PRRT3、STEAP2、GPAT3、KRT32、ABI3BP、SFRP4、SFRP4等14个 miRNA-708 相关靶基因；靶向沉默FKBP52可降低miRNA-708在BMSCs中的表达；激活FKBP52/GRα/miRNA-708信号通路可抑制BMSCs成骨分化、促进其成脂分化，导致SANFH的发生；FKBP52/GRα/miRNA-708信号通路在BMSCs的成骨-成脂分化过程中起到关键作用，生骨再造丸靶向调控FKBP52/GRα/miRNA-708信号通路是治疗SANFH的主要机制之一。