Osteoporosis is a bone metabolic disease caused by the imbalance between osteoblasts and osteoclasts due to excess osteoclastogenesis, manifesting in the decrease of bone density and bone strength. Scutellariae Radix shows good anti-osteoporosis activity, but the effective component is still unclear. Cell membrane chromatography (CMC) is a biological affinity chromatography with membrane immobilized on a silica carrier as the stationary phase. It can realize a dynamical simulation of interactions between drugs and receptors on cell membrane, which is suitable for screening active compounds from complex systems. In this study, the components of Scutellariae Radix with potential anti-osteoporosis activity through inhibiting the differentiation from bone marrow mononuclear cells (BMMCs) to osteoclast were screened by a BMMC/CMC analytical system. Firstly, a new 3-mercaptopropyltrimethoxysilane (MPTS)-modified BMMC/CMC stationary phase was developed to realize covalent binding with cell membrane fractions. By investigating the retention time (t_R) of the positive drug, the life span of the MPTS-modified CMC columns was significantly improved from 3 to 12 days. Secondly, 6 components of Scutellariae Radix were screened to show affinity to membrane receptors on BMMCs by a two-dimensional BMMC/CMC–TOFMS analytical system. Among them, tectochrysin demonstrated the best anti-osteoporosis effect in vitro, which has never been reported. We found that tectochrysin could inhibit the differentiation of BMMCs into osteoclasts induced by receptor activator of nuclear factor-κΒ ligand (RANKL) and macrophage colony-stimulating factor (M-CSF) in a concentration-dependent manner in vitro. In vivo, it significantly reduced the loss of bone trabeculae in ovariectomized mice, and decreased the level of C-terminal cross-linking telopeptides of type 1 collagen (CTX-1), tartrate-resistant acid phosphatase 5b (TRAP-5b), interleukin 6 (IL-6) in serum. In conclusion, tectochrysin serves as a potential candidate in the treatment of osteoporosis. The proposed two-dimensional MPTS-modified BMMC/CMC-TOFMS analytical system shows the advantages of long-life span and fast recognition ability, which is very suitable for infrequent cell lines.

骨质疏松症是一种骨代谢疾病，是由于过度的破骨细胞形成导致的成骨细胞与破骨细胞之间的不平衡所致，表现为骨密度和骨强度的降低。黄cut显示出良好的抗骨质疏松活性，但有效成分仍不清楚。细胞膜色谱法（CMC）是一种生物亲和色谱法，其膜固定在硅胶载体上作为固定相。它可以实现药物与细胞膜上受体之间相互作用的动力学模拟，适用于从复杂系统中筛选活性化合物。在这项研究中，通过BMMC / CMC分析系统筛选了通过抑制从骨髓单核细胞（BMMC）向破骨细胞分化而具有潜在抗骨质疏松活性的黄S成分。首先，开发了一种新的3-巯基丙基三甲氧基硅烷（MPTS）修饰的BMMC / CMC固定相，以实现与细胞膜级分的共价结合。通过调查保留时间（（t _R）为阳性药物，MPTS修饰的CMC色谱柱的使用寿命从3天缩短到12天。其次，通过二维BMMC / CMC-TOFMS分析系统筛选了黄S的6个成分，以显示对BMMC膜受体的亲和力。其中，线粒体蛋白在体外显示出最佳的抗骨质疏松作用，但尚未见报道。我们发现，tectochrysin可抑制的BMMCs分化成由核因子的受体活化诱导的破骨细胞κ以依赖于浓度的方式Β配体（RANKL）和巨噬细胞集落刺激因子（M-CSF）在体外。体内，可显着减少卵巢切除小鼠的骨小梁丢失，并降低1型胶原（CTX-1），抗酒石酸酸性磷酸酶5b（TRAP-5b），白介素6（ IL-6）在血清中。总之，血凝素可以作为骨质疏松症的潜在治疗药物。提出的二维MPTS修饰的BMMC / CMC-TOFMS分析系统显示出长寿命和快速识别的优势，非常适合于不常见的细胞系。