基于PI3K/AKT信号通路PHB调节PMOP成骨细胞凋亡作用及补肾活血方干预机制的研究

Abnormal apoptosis in postmenopausal osteoblast is another important mechanism of postmenopausal osteoporosis. Prohibitin (PHB) plays an important role in the regulation of cell proliferation and cell apoptosis. PI3K/Akt pathway is a pathway in regulating osteoblast apoptosis, and is associated with ERα. Although PHB is associated to the PI3K/Akt pathway, there is no report on the mechanism of PHB regulating osteoblast apoptosis at present. We found that Bushen Huoxue Recipe could cause the different expression of PHB in osteoblast, with PHB expression increasing and Akt level lowing. Thus we hypothesized that PHB is the major target of Bushen Huoxue Recipe and it may through the pathway of PI3K/AKT to regulate osteoblastic cell apoptosis. In this project,we intend to construct PHB overexpression and silencing the expression of osteoblastic cell model to observe the effect of PHB expression changes on PI3K/AKT pathway、ER and mitochondria pathway key factors in the osteoblast cells, which researches the influence of PHB on the regulation of apoptosis in osteoblast cells; through the vivo experimentes, we could observe the effect of Bushen Huoxue Recipe on PHB、ER 、regulated proteins and cytokines in the PMOP rats bone tissue to investigate the action mechanism of Bushen Huoxue Recipe in the expression of PMOP rats bone tissue PHB and the regulation of oapoptosis in osteoblastic cell. Thus, this is to provide a new theoretical basis for the cause of PMOP and to provide a new target and path for the prevention and treatment of PMOP.

绝经后OB的异常凋亡是PMOP的又一重要机制。PHB在细胞凋亡的调控中起重要作用。PI3K /Akt通路参与调控成骨细胞凋亡且与ERα相关，虽然PHB和PI3K /Akt密切相关，但尚未见PHB调节OB凋亡机制的报道。我们在前期研究中发现黄芪三仙汤能引起PMOP成骨细胞PHB的差异表达，且PHB表达量升高，Akt水平下降。我们推测PHB是黄芪三仙汤的关键作用靶点，可能通过PI3K/AKT通路来调节OB凋亡。本研究拟构建PHB过表达和沉默表达OB模型，观察PHB表达变化对OB的ER、PI3K/AKT通路等的影响，探讨PHB对调节OB凋亡的作用机制；通过体内实验，观察黄芪三仙汤对PMOP大鼠骨组织PHB、ER、相关蛋白等的影响，探讨其对PMOP大鼠骨组织PHB表达的影响及其调节骨细胞凋亡的作用机制。为PMOP病因提供新的理论依据，为中医药防治PMOP提供新的作用靶点及路径。

绝经后成骨细胞的异常凋亡是PMOP的又一重要机制。PHB在细胞凋亡的调控中起重要作用。PI3K/Akt通路参与调控成骨细胞凋亡相关，虽然PHB和PI3K/Akt密切相关，但尚未见PHB调节OB凋亡机制的报道。本研究通过构建PHB过表达和沉默病毒感染PMOP OB模型，证实了PHB是PMOP的一个重要的影响因子，也是药物防治PMOP的一个重要作用靶点。高表达或低表达PHB能通过PI3K/Akt信号通路以调节PMOP OB的成骨分化及矿化结节并调控凋亡相关蛋白。并通过体内和体外实验证实了补肾活血方可通过抑制PHB的表达，激活PI3K/Akt信号通路，促进PMOP OB的成骨分化，抑制PMOP OB的细胞凋亡，从而发挥防治PMOP的作用，部分阐明了补肾活血方防治PMOP的作用机制，为进一步的深入研究打下坚实基础。