Abstract：
Prolyl 4-hydroxylase beta subunit (P4HB) plays a vital role in bone formation. This study intends to clarify the role of P4HB in the therapeutic effect of Icariin (ICA) on osteoporosis. Herein, in vivo and in vitro models were constructed by performing ovariectomy (OVX) in rats and inducing osteogenic differentiation in bone marrow stem cells (BMSCs), respectively. Hematoxylin and eosin staining and micro-computed tomography analysis were performed to evaluate osteoporosis in OVX rats. Alizarin Red staining, alkaline phosphatase staining, and the ALP activity test were employed to assess osteogenesis. m6A dot blotting and methylated RNA immunoprecipitation were used to determine m6A modification. We found that P4HB was downregulated in bone tissues of patients with osteoporosis and OVX rats. P4HB facilitated osteogenic differentiation of BMSCs. What\'s more, ICA upregulated P4HB expression, promoted osteogenic differentiation of BMSCs, and alleviated osteoporosis in OVX rats, which were reversed by knocking down P4HB. ICA enhanced the stability and m6A modification of P4HB. METTL14 mediated m6A modification of P4HB mRNA. In addition, METTL14 knockdown overturned the promotive effects of ICA on P4HB m6A level and BMSC osteogenic differentiation. To sum up, ICA elevated the METTL14-mediated m6A modification of P4HB to facilitate BMSC osteogenic differentiation.
摘要：
脯氨酸4-羟化酶β亚基（P4HB）在骨形成中起着至关重要的作用。本研究旨在阐明P4HB在淫羊藿苷(ICA)治疗骨质疏松症中的作用。在这里，通过在大鼠中进行卵巢切除术（OVX）和诱导骨髓干细胞（BMSCs）成骨分化来构建体内和体外模型，分别。进行苏木精和伊红染色和显微计算机断层扫描分析以评估OVX大鼠的骨质疏松症。茜素红染色,碱性磷酸酶染色，和ALP活性测试用于评估成骨。m6A斑点印迹和甲基化RNA免疫沉淀用于确定m6A修饰。我们发现P4HB在骨质疏松症患者和OVX大鼠的骨组织中下调。P4HB促进BMSCs成骨分化。更重要的是，ICA上调P4HB表达，促进BMSCs成骨分化，减轻OVX大鼠的骨质疏松症，通过击倒P4HB来逆转。ICA增强了P4HB的稳定性和m6A修饰。METTL14介导m6A修饰P4HBmRNA。此外,METTL14敲低推翻了ICA对P4HBm6A水平和BMSC成骨分化的促进作用。总而言之,ICA提高了METTL14介导的P4HB的m6A修饰以促进BMSC成骨分化。
