cAMP/PKA/CREB信号通路对卵巢早衰继发骨质疏松大鼠骨微结构影响研究
Effect of cAMP/PKA/CREB signaling pathway on bone microstructure in rats with osteoporosis secondary to premature ovarian failure
张雁儒 1杨越 2董家琪 2徐景超 2韩道正 2苏甲子2
作者信息
- 1. 宁波大学医学部,浙江宁波 315211
- 2. 河南理工大学骨科研究所,河南 焦作 454000
- 折叠
摘要
目的 探究环磷酸腺苷(cyclicadenosine monophosphate,cAMP)/蛋白激酶A(protein kinase A,PKA)/cAMP反应元件结合蛋白(cAMP response element binding protein,CREB)信号通路在卵巢早衰继发骨质疏松疾病进程中对大鼠骨微结构及骨髓间充质干细胞成骨矿化结节活力影响.方法 利用生物信息方法分析cAMP/PKA/CREB信号通路;对成年雌性大鼠腹腔注射环磷酰胺建立卵巢早衰继发骨质疏松疾病模型,首次给药剂量50mg/kg,后续8mg/kg为期两周设为模型组,同时设生理盐水组、老龄组进行对照,建模后腹腔注射1mg/kgPKA抑制剂H89为期两周;取大鼠股骨头、骨髁进行切片HE、番红固绿染色;取三组大鼠骨髓间充质干细胞进行成骨诱导茜素红染色.结果 环磷酰胺成功诱导雌鼠卵巢早衰继发骨质疏松.当PKA被抑制后,干细胞成骨矿化结节活力明显降低,三组骨微结构均呈明显下降,主要表现在骨骺、骺板的软骨增殖区和软骨钙化区以及骨小梁,生理盐水组最为明显.结论 cAMP/PKA/CREB信号通路在调控卵巢颗粒细胞生理活动、成骨分化及软骨生长方面有着重要意义,当抑制PKA后,大鼠骨微结构呈明显下降.
Abstract
Objective To explore cyclicadenosine monophosphate(cAMP)/protein kinase A(PKA)/cAMP response element binding effect of proteins protein,CREB)signaling pathway in the process of osteoporosis secondary to premature ovarian failure on bone microstructure and bone marrow mesenchymal stem cells mineralized nodules activity in rats.Methods Bioinformatics methods were used to analyze the cAMP/PKA/CREB signaling pathway.The model of osteoporosis secondary to premature ovarian failure was established by intraperitoneal injection of cyclophosphamide(50mg/kg for the first time,followed by 8mg/kg for two weeks)in adult female rats.At the same time,the normal saline group and the aged group were set as controls,and 1mg/kg PKA inhibitor H89 was injected intraperitoneally for two weeks after modeling.The femoral head and condyle were stained with HE and Safranin fast green.The bone marrow mesenchymal stem cells of the three groups were stained with osteogenic induction alizarin red.Results Cyclophosphamide successfully induced osteoporosis secondary to premature ovarian failure in female rats.When PKA was inhibited,the activity of bone mineralized nodules of stem cells was significantly decreased,and the bone microstructure of the three groups was significantly decreased,mainly in the epiphysis,the cartilage proliferation area and cartilage calcification area of the epiphyseal plate,and the trabecular bone,especially in the saline group.Conclusions The cAMP/PKA/CREB signaling pathway plays an important role in regulating the physiological activities of ovarian granulosa cells,osteogenic differentiation and cartilage growth.Inhibition of PKA significantly decreases the bone microstructure in rats.
关键词
cAMP/PKA/CREB/卵巢早衰继发骨质疏松/骨微结构Key words
cAMP/PKA/CREB/Osteoporosis secondary to premature ovarian failure/Bone microstructure引用本文复制引用
基金项目
河南省科技攻关(172102310546)
出版年
2024
NETL
NSTL
万方数据