运用CRISPR/CAS9敲除斑马鱼hepcidin基因来研究铁蓄积对绝经后骨质疏松的影响及其机制

In recent years, studies have shown that postmenopausal osteoporosis occur not only due to estrogen deficiency, but also owing to the accumulation of body iron content during perimenopausal and menopausal period. There are some reports which have confirmed the efficacy of some way to reduce the body iron content in preventing and treating postmenopausal osteoporosis. .However, the concrete mechanism of iron accumulation induced abnormal bone metabolism is still unclear,In this project, We observe the hepcidin-/- zebrafish by CRISPR/CAS9 through detecting a series of markers. These markers include iron metabolism markers,bone metabolism markers,bone mineral density and bone metabolism genes.We also cultivate primitive cells in vitro to study the effect of iron accumulation on BMP2 signaling pathways.This study may provide the new evidence to explain the abnormal bone metabolism induced by iron accumulation, and thus explore the therapeutical way based on the findings.

近年来的研究表明，绝经后骨质疏松症（Ⅰ型骨质疏松症）的发生不仅与绝经后雌激素缺乏有关，而且与绝经后体内“铁蓄积”有关。目前已有报道证实：“铁蓄积”抑制骨形成，促进骨质疏松，且“降低体内铁含量方法”具有防治Ⅰ型骨质疏松症的疗效。.然而，铁蓄积对骨代谢的影响机制尚不明确。本项目运用“CRISPR/ CAS9技术”敲除斑马鱼铁调素基因，检测转基因斑马鱼铁代谢指标、骨代谢指标、骨密度及相关的骨代谢基因；并利用原代细胞，研究“BMP2a—Runx2a信号通路”指标改变，最终通过实验，探讨“铁蓄积”导致骨代谢异常的影响和机制，希望能以此项研究，提供应对“Ⅰ型骨质疏松症”的防治新策略。

近年来的研究表明，绝经后骨质疏松症（Ⅰ型骨质疏松症）的发生不仅与绝经后雌激素缺乏有关，而且与绝经后体内“铁蓄积”有关。目前已有报道证实：“铁蓄积”抑制骨形成，促进骨质疏松，且“降低体内铁含量方法”具有防治Ⅰ型骨质疏松症的疗效。本项目运用CRISPR/CAS9技术敲除斑马鱼铁调素基因，检测转基因斑马鱼铁代谢指标、骨代谢指标、骨密度及相关的骨代谢基因，发现敲除斑马鱼铁调素基因造成斑马鱼的铁过载，骨密度下降，骨代谢基因下降，从高通量测序结果发现hepcidin通过调控BMP2a的启动子来调控的Runx2a促进骨骼的发育，并通过定量和Westernblot实验来证实。初步发现“铁蓄积”导致骨代谢异常的影响和机制，为“Ⅰ型骨质疏松症”的防治新策略。