摘要
目的文章对刺老苞根皮进行化学成分及其对原代破骨细胞影响研究。方法采用大孔吸附树脂、硅胶柱、ODS-C_(18)以及制备型HPLC等方法进行分离纯化,结合理化性质和波谱数据鉴定化合物结构。体外培养原代破骨细胞,采用抗酒石酸酸性磷酸酶阳性染色及骨吸收甲苯胺蓝染色进行鉴定。CCK-8细胞毒性检测法、TRAP阳性细胞数实验及Real-time PCR(RT-PCR)与Western blot法测定骨保护素(osteoprotegerin,OPG)及核因子-κB受体活化因子配体(receptor activator of nuclear factor kappaB ligand,RANKL)的表达情况检测化合物对破骨细胞的影响。结果成功从刺老苞根皮70%乙醇提取物中分离鉴定了4个化合物,分别是:屏边三七皂苷R_(2)、楤木皂苷C、楤木皂苷A、竹节人参皂苷IVa。在0~50μg·mL^(-1)浓度范围可排除化合物对破骨细胞抑制的假阳性结果。与对照组相比,4个化合物低、中、高浓度(0.5μg·mL^(-1)、5μg·mL^(-1)、50μg·mL^(-1))组OPG mRNA与蛋白表达有所升高,RANKL mRNA与蛋白表达有所降低,破骨细胞数目均有所减少,数据有统计学差异意义(P<0.05或P<0.01)。结论从刺老苞根皮中分离得到的4个单体化合物对原代破骨细胞有一定的抑制活性,为刺老苞根皮治疗骨质疏松症的物质基础研究提供依据。
Objective This paper was aimed to investigate the chemical constituents from Aralia echinocaulis Hand.-Mazz.,as well as their effects on primary osteoclasts.Methods The constituents were isolated and purified by column chromatography over macroporous resin,silica gel,ODS-C_(18),and preparative HPLC.Their structures were identified by spectra analysis.Primary osteoclasts were cultured in vitro,and identified by anti-tartaric acid phosphatase positive staining and bone resorption toluidine blue staining.CCK-8 cytotoxicity assay,TRAP positive cell count assay,the expression mRNA and protein levels of OPG/RANKL were used to test the effect of constituents on osteoclasts.Results The results suggested that four compounds were obtained from 70%ethanol extract of Aralia echinocaulis Hand.-Mazz.and identified as Stipuleanoside R_(2),Araloside C,Araloside A,Chikuselsusaponin IVa.In the range of 0~50μg·mL^(-1),the false positive result of inhibition of osteoclasts by monomeric compounds can be excluded.Low,medium,and high concentrations of four compounds(0.5μg·mL^(-1),5μg·mL^(-1),50μg·mL^(-1))showed a decrease in the number of osteoclasts and the expression mRNA and protein levels of RANKL,and a increase in the expression mRNA and protein levels of OPG,compared with the control group,indicating a certain inhibitory effect.There was statistically significant difference(P<0.05 or P<0.01).Conclusion These findings indicated that four compounds,which were isolated from Aralia echinocaulis Hand.-Mazz.,had certain inhibitory activity on primary osteoclasts,which provided a basis for the material basic research of the treatment of osteoporosis.
作者
燕梦云
李金诚
周文斌
裴凌鹏
YAN Meng-yun;LI Jin-cheng;ZHOU Wen-bin;PEI Ling-peng(Key Laboratory of Ethnomedicine,Miruu University of China,Ministry of Education,Beijing 100081,China;Xinjiang Teachers College,Urumqi,Xinjiang 830043,China)
出处
《时珍国医国药》
CAS
CSCD
北大核心
2021年第8期1793-1797,共5页
Lishizhen Medicine and Materia Medica Research
基金
国家自然科学基金(81673769)。
