Development of 3-mercaptopropyltrimethoxysilane (MPTS)-modified bone marrow mononuclear cell membrane chromatography for screening anti-osteoporosis components from Scutellariae Radix.
Development of 3-mercaptopropyltrimethoxysilane (MPTS)-modified bone marrow mononuclear cell membrane chromatography for screening anti-osteoporosis components from Scutellariae Radix.
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3-巯基丙基三甲氧基硅烷（MPTS）修饰的骨髓单核细胞膜色谱法筛选黄芩抗骨质疏松成分的研究
DOI：
10.1016/j.apsb.2020.01.019
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发表时间：
2020-10
期刊：
影响因子：
--
通讯作者：
Chen X
中科院分区：
文献类型：
--
作者：
Gu Y;Chen X;Wang Y;Liu Y;Zheng L;Li X;Wang R;Wang S;Li S;Chai Y;Su J;Yuan Y;Chen X
骨质疏松症是由于破骨细胞过度生成，导致成骨细胞和破骨细胞之间的失衡，表现为骨密度和骨强度降低而引起的一种骨代谢疾病。黄芩具有良好的抗骨质疏松活性，但其有效成分尚不清楚。细胞膜色谱(CMC)是一种以硅胶载体为固定相的生物亲和层析。它可以在细胞膜上实现药物与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。本研究利用BMMC/CMC分析系统筛选出具有抑制骨髓单个核细胞向破骨细胞分化而具有抗骨质疏松作用的有效成分。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，实现了与细胞膜部分的共价结合。通过考察阳性药物的保留时间，MPTS修饰的CMC柱的寿命从3天提高到12天。其次，利用双向BMMC/CMC-TOFMS分析系统，筛选出6个与BMMCs上的膜受体具有亲和力的成分。其中，ectochrysin在体外表现出最好的抗骨质疏松症作用，这是从未报道过的。结果表明，延胡索素在体外对核因子受体激活剂κΒ配体和巨噬细胞集落刺激因子诱导的骨髓基质细胞向破骨细胞的分化有抑制作用，且呈浓度依赖性。在体内可明显减少去卵巢小鼠骨小梁的丢失，降低血清中C端交联型胶原末端肽(CTX-1)、抗酒石酸酸性磷酸酶5b(TRAP-5b)、白介素6(IL-6)水平。综上所述，tectochrysin有望成为治疗骨质疏松症的候选药物。所提出的MPTS修饰的二维BMMC/CMC-TOFMS分析系统具有寿命长、识别能力快的优点，非常适合于罕见的细胞系。为了筛选黄芩中抗骨质疏松的成分，建立了2D BMMC/CMC-TOFMS分析体系，并对硅胶固定相进行MPTS修饰，显著延长了色谱柱寿命。首次筛选出具有良好抗骨质疏松活性的长春花素。
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 (tR) 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. To screen anti-osteoporosis components from Scutellariae Radix, 2D BMMC/CMC-TOFMS analytical system was established and the column life span was significantly prolonged by MPTS modification on silica stationary phase. Tectochrysin was screened out with good anti-osteoporosis activity for the first time.
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DOI：
10.1096/fj.201700316r
发表时间：
2017-11
期刊：
FASEB journal : official publication of the Federation of American Societies for Experimental Biology
影响因子：
--
作者：
Chen X;Zhi X;Pan P;Cui J;Cao L;Weng W;Zhou Q;Wang L;Zhai X;Zhao Q;Hu H;Huang B;Su J
通讯作者：
Su J
影响因子：
9
作者：
Chen X;Zhi X;Cao L;Weng W;Pan P;Hu H;Liu C;Zhao Q;Zhou Q;Cui J;Su J
通讯作者：
Su J
全面的二维 APTES 修饰的 MCF7 细胞膜色谱系统用于表征元胡-白芷草药对中潜在的抗乳腺癌成分
影响因子：
3.6
作者：
Wang, Xiao-Yu;Ding, Xuan;Yuan, Yong-Fang;Zheng, Le-Yi;Cao, Yan;Zhu, Zhen-Yu;Zhang, Guo-Qing;Chai, Yi-Feng;Chen, Xiao-Fei;Hong, Zhan-Ying
通讯作者：
Hong, Zhan-Ying
影响因子：
4.2
作者：
de Juan-Franco, Elena;Caruz, Antonio;Lechuga, Laura M.
通讯作者：
Lechuga, Laura M.
DOI：
10.1016/j.jpba.2018.10.028
发表时间：
2019-02-05
影响因子：
3.4
作者：
Gu, Yanqiu;Chen, Xiaofei;Yuan, Yongfang
通讯作者：
Yuan, Yongfang