中国组织工程研究2025，Vol.29Issue(32)：6828-6835,8.DOI:10.12307/2025.970
健骨方对破骨细胞-成骨细胞的偶联效应及作用机制
Mechanism of Jiangu Formula in treating osteoporosis based on osteoclast-osteoblast coupling
摘要
Abstract
BACKGROUND:Previous studies have found that Jiangu Formula can play a dual role in anti-osteoporosis therapy by inhibiting bone resorption and promoting bone formation,but its mechanism of action has not been elucidated. OBJECTIVE:To investigate the effect of Jiangu Formula on the coupling mechanism between bone resorption and bone formation at the cellular and molecular levels. METHODS:Serum containing Jiangu Formula was prepared and its toxicity to bone marrow derived macrophages was determined using MTT assay.The cell proliferation ability of MC3T3-E1 cells that treated Jiangu Formula medicated serum was measured by using cell counting kit-8 method.Tartrate resistant acid phosphatase staining was used to determine the bone resorption inhibition effect of serum containing Jiangu Formula on osteoclast differentiation.The Jiangu Fang conditional culture medium was prepared and the promoting effect of the conditional culture medium on bone formation of MC3T3-E1 was determined through alkaline phosphatase staining.The effect of serum containing Jiangu Formula on the mRNA levels of osteoclast associated sphingosine kinase 2,collagen triple helix repeat containing 1,and slit homolog 3 coupled genes,as well as the effect of conditioned culture medium on the mRNA levels of bone formation related secreted phosphoprotein 1,alkaline phosphatase,Sp7 transcription factor,and secreted protein acidic and rich in cysteine genes,were determined by RT-qPCR method. RESULTS AND CONCLUSION:(1)The MTT and cell counting kit-8 methods showed that serum containing Jiangu Formula had no significant cytotoxic effects at a concentration of<5%.(2)Tartrate resistant acid phosphatase staining results indicated that serum containing Jiangu Formula dose-dependently inhibited osteoclast differentiation at 2.5%,1.25%,and 0.63%concentrations.(3)The alkaline phosphatase staining results showed that the osteoclast conditioned medium of Jiangu Formula promoted osteoblast differentiation in a dose-dependent manner.(4)In addition,RT-qPCR results showed that serum containing 0.63%Jiangu Formula could increase the mRNA levels of osteoclast derived slit homolog 3 and sphingosine kinase 2 coupling factors,and the conditional culture medium could increase the mRNA expression levels of bone formation related secreted phosphoprotein 1,Sp7 transcription factor,and secreted protein acidic and rich in cysteine.To conclude,the serum containing Jiangu Formula can promote bone formation while inhibiting bone resorption,and its mechanism may be related to the mRNA expression of the coupling factors osteoclast derived slit homolog 3 and sphingosine kinase 2 that promote bone resorption and bone formation.关键词
健骨方/骨吸收/骨形成/骨质疏松症/偶联/破骨细胞/成骨细胞/骨稳态/工程化组织构建Key words
Jiangu Formula/bone resorption/bone formation/osteoporosis/coupling/osteoclasts/osteoblasts/bone homeostasis/engineered tissue construction分类
医药卫生引用本文复制引用
卢秀丽,许华珍,陈玉兴,姚楠,胡子旋,黄丹娥..健骨方对破骨细胞-成骨细胞的偶联效应及作用机制[J].中国组织工程研究,2025,29(32):6828-6835,8.基金项目
国家自然科学基金面上项目(82104473),项目负责人:黄丹娥 （82104473）
国家自然科学基金青年科学基金项目(82474347),项目负责人:黄丹娥 （82474347）
广东省自然科学基金(2023A1515010752),项目负责人:黄丹娥 （2023A1515010752）
广东省第二中医院科研创新基金项目(SEZYY2023A02),项目负责人:姚楠 National Natural Science Foundation of China(General Program),No.82104473(to HDE) （SEZYY2023A02）
National Natural Science Foundation of China(Youth Program),No.82474347(to HDE) （Youth Program）
Natural Science Foundation of Guangdong Province,No.2023A1515010752(to HDE) （to HDE）
Guangdong Second Hospital of Traditional Chinese Medicine Scientific Research and Innovation Fund Project,No.SEZYY2023A02(to YN) （to YN）
