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大鼠胆汁中黄芩苷代谢物的分离、纯化及其对肝癌细胞HepG2增殖的影响研究
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| 篇名： | 大鼠胆汁中黄芩苷代谢物的分离、纯化及其对肝癌细胞HepG2增殖的影响研究 |
| TITLE： | Study on the Isolation and Purification of Baicalin Metabolites from Rat Bile and Its Effect on the Proliferation of Liver Cancer HepG 2 Cells |
| 摘要： | 目的：从大鼠胆汁中分离纯化黄芩苷的代谢物，并研究其对肝癌细胞HepG2增殖的影响。方法：取6只SD大鼠，麻醉后进行胆管插管，待大鼠清醒后，灌胃给予黄芩苷（168mg/kg），收集灌胃给药后24h的胆汁样品1，经处理后，采用高效液相色谱法进行初步分析；另取5只SD大鼠，同上述方法麻醉、插管后，灌胃给予黄芩苷（400mg/kg），并收集灌胃给药后48h的胆汁样品2，经处理后，采用半制备型高效液相色谱法对代谢物进行分离纯化；采用紫外光谱、红外光谱、质谱、核磁共振技术并结合其理化性质对分离得到的代谢物进行鉴定；采用MTT法及高内涵细胞成像分析法研究代谢物对HepG2细胞增殖的影响。结果：黄芩苷通过胆汁主要代谢成代谢物1和代谢物2，经分离纯化后分别鉴定为千层纸素A-7-O-β-D-葡萄糖醛酸苷和黄芩素6-O-β-D-葡萄糖醛酸苷。MTT法结果显示，代谢物2对HepG2细胞具有显著的增殖抑制作用，其半数抑制浓度（IC50）为90µg/mL；高内涵细胞成像分析法结果显示，在一定浓度下，代谢物2可能通过改变HepG2细胞线粒体分布及膜通透性从而抑制细胞增殖（代谢物1的药理活性已有报道，本研究略去）。结论：成功分离并鉴定出2个黄芩苷的胆汁代谢物，其中代谢物2黄芩素6-O-β-D-葡萄糖醛酸苷对HepG2细胞具有显著的增殖抑制作用。 |
| ABSTRACT： | OBJECTIVE：To isolate and purify baicalin metabolites from rat bile ，and to study its effect on the proliferation of liver cancer HepG 2 cells. METHODS ：Totally of 6 SD rats were collected ，anesthetized and then intubated with bile ducts. They were given baicalin （168 mg/kg）intragastrically after the rats were awake as well as the bile sample 1 was collected during 24 h after intragastric administration. After processed ，bile sample 1 was preliminarily analyzed by HPLC. Another 5 SD rats were anesthetized and intubated in the same way as above ，and then given baicalin intragastrically （400 mg/kg）. The bile sample 2 was collected during 48 h after intragastric administration. After processed ，the bile sample 2 was isolated and purified by semi-preparative HPLC. The isolated metabolites were identified by using UV ，IR，MS and NMR ，as well as based on physical and chemical properties. MTT method combined with high content cell imaging analysis system was used to study the effect of the metabolites on the proliferation of HepG 2 cells. RESULTS ：Baicalin was mainly metabolized into metabolite 1 and metabolite 2 through bile. After isolation and purification ， they were identified as oroxylin A- 7-O-β-D-glucuronide and 2726719792@qq.com baicalein 6-O-β-D-glucuronide. The results of MTT assay showed that the metabolite 2 had a significant inhibitory effect 分析。E-mail：gaoxl@gmc.edu.cn on the proliferation of HepG 2 cells，and its IC 50 was 90 µg/mL；the results of high content cell imaging analysis showed that at a certain concentration metabolite 2 may inhibit proliferation by changing the mitochondrial distribution and membrane permeability of HepG 2 cells（studies on the pharmacological activities of metabolism 1 had been reported ，it was skipped in this study ）. CONCLUSIONS ：In this study ，two baicalin bile metabolites were successfully isolated and identified ，of which baicalein 6-O-β-D-glucuronide has a significant inhibitory effect on the proliferation of HepG 2 cells. |
| 期刊： | 2020年第31卷第07期 |
| 作者： | 朱亚南，张硕，张敏，孟小夏，杨七妹，汤磊，张荣平，高秀丽 |
| AUTHORS： | ZHU Yanan， ZHANG Shuo，ZHANG Min，MENG Xiaoxia，YANG Qimei，TANG Lei，ZHANG Rongping，GAO Xiuli |
| 关键字： | 黄芩苷；代谢物；千层纸素A-7-O-β-D-葡萄糖醛酸苷；黄芩素6-O-β-D-葡萄糖醛酸苷；分离纯化；高内涵细胞成像分析；肝 |
| KEYWORDS： | Baicalin； Metabolite； Oroxylin A- 7-O-β-D-glucuronide； Baicalein 6-O-β-D-glucuronide； Isolation and |
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