Gu Yanqiu, Chen Xiao, Wang Yao, Liu Yue, Zheng Leyi, Li Xiaoqun, Wang Rong, Wang Shaozhan, Li Shengnan, Chai Yifeng, Su Jiacan, Yuan Yongfang, Chen Xiaofei
Department of Pharmacy, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University, School of Medicine, Shanghai 201999, China.
School of Pharmacy, Second Military Medical University, Shanghai 200433, China.
Acta Pharm Sin B. 2020 Oct;10(10):1856-1865. doi: 10.1016/j.apsb.2020.01.019. Epub 2020 Feb 18.
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 ( ) 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 , which has never been reported. We found that tectochrysin could inhibit the differentiation of BMMCs into osteoclasts induced by receptor activator of nuclear factor-Β ligand (RANKL) and macrophage colony-stimulating factor (M-CSF) in a concentration-dependent manner . , 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.
骨质疏松症是一种由于破骨细胞生成过多导致成骨细胞与破骨细胞之间失衡而引起的骨代谢疾病，表现为骨密度和骨强度降低。黄芩具有良好的抗骨质疏松活性，但其有效成分仍不清楚。细胞膜色谱法（CMC）是以固定在硅胶载体上的膜为固定相的一种生物亲和色谱法。它可以实现药物与细胞膜上受体相互作用的动态模拟，适用于从复杂体系中筛选活性化合物。在本研究中，通过骨髓单核细胞（BMMC）/CMC分析系统筛选了黄芩中具有潜在抗骨质疏松活性、能抑制骨髓单核细胞向破骨细胞分化的成分。首先，开发了一种新型的3-巯基丙基三甲氧基硅烷（MPTS）修饰的BMMC/CMC固定相，以实现与细胞膜组分的共价结合。通过考察阳性药物的保留时间（ ），MPTS修饰的CMC柱的寿命从3天显著提高到12天。其次，通过二维BMMC/CMC-TOFMS分析系统筛选出黄芩的6种成分，显示出它们与BMMC上的膜受体具有亲和力。其中，黄芩黄酮表现出最佳的抗骨质疏松效果，这从未有过报道。我们发现黄芩黄酮能以浓度依赖的方式抑制核因子-κB受体活化因子配体（RANKL）和巨噬细胞集落刺激因子（M-CSF）诱导的BMMC向破骨细胞的分化。此外，它显著减少了去卵巢小鼠骨小梁的丢失，并降低了血清中I型胶原C端交联肽（CTX-1）、抗酒石酸酸性磷酸酶5b（TRAP-5b）、白细胞介素6（IL-6）的水平。总之，黄芩黄酮是治疗骨质疏松症的潜在候选药物。所提出的二维MPTS修饰的BMMC/CMC-TOFMS分析系统具有寿命长和识别速度快的优点，非常适合于不常用的细胞系。
