药物血清内黄芪甲苷含量测定及其抑制HSCs活化增殖的实验研究(1)
摘 要：目的：探讨黄芪经正常、肝纤维化机体代谢后血清内黄芪甲苷含量差别，及其对肝纤维化发生中HSCs活化增殖的影响。方法：40%CCl4, 皮下注射9周制备大鼠肝纤维化模型。正常、肝纤维化大鼠分别灌服黄芪，提取正常、肝纤维大鼠药物血清，HPLC检测药物血清内黄芪甲苷含量。CCK-8法检测黄芪甲苷对HSCs增殖影响，3-Pro掺入-胶原酶消化法检测黄芪甲苷对HSCs胶原分泌的影响。结果：正常、病理药物血清内均可检出黄芪甲苷。病理药物血清内黄芪甲苷含量高于正常药物血清(0.1140 vs0.0851,P<0.05 )。黄芪甲苷可抑制HSCs增殖及I型胶原分泌。结论：机体状态可影响黄芪内有效成分黄芪甲苷的代谢，可能与肝脏不同状态、口服药物首过效应及分解代谢有关。HPLC可用于检测中药体内有效成分含量，评估机体代谢对于中药有效成分含量的影响及临床药效。黄芪甲苷可有效抑制HSCs增殖、活化，为抗肝纤维化有效作用成分之一。
关键词：黄芪甲苷；血清药理学；高效液相色谱法；肝纤维化；肝星状细胞
, http://www.100md.com
中图分类号：R285.5文献标识码：A文章编号：1673-7717(2011)12-2770-04
Content Assaying of Astragaloside IV and Its Effects on Proliferation and Activation of
HSCs in Liver Fibrosis
LU Tao1, YAO Xixian2, SUN Zeming2
(1. Dapartment of Gastroenterology, First People's Hospital of Hangzhou, Hangzhou 310006, Zhejiang, China;
2. Hebei Institute of Gastroenterology, Shijiazhuang 050000, Hebei, China)
, 百拇医药
________________________________________
收稿日期：2011-07-10
作者简介：吕涛(1976-)，女，河北石家庄人，医师，博士，研究方向：中药防治肝纤维化作用机制及成分研究。Abstract:Objective: To detect the contents of astragaloside Ⅳ in raw astragalus mongholicus, and the corresponding drug sera (normal and fibrotic drug sera) by HPLC. To compare the astragaloside Ⅳ in astragalus mongholicus. To study the effects of astragaloside Ⅳon proliferation and activation of HSCs in hepatic fibrogenesis. Methods: Inducing liver fibrosis in rats by 40% CCl4, SC, for 9 weeks. Astragalus mongholicus. solutions were given to the normal and fibrotic rats, respectively for 5days. Normal and fibrotic drug sera from rats were extracted. The separation of astragaloside Ⅳwas performed by HPLC. The effects of astragaloside Ⅳ on the proliferation of HSCs were measured by the method of CCK-8 (Cell Counting Kit-8). The effects on synthesis of collagen I were assessed by 3H-proline incorporation assays. Results: Astragaloside Ⅳ could be found both in normal and fibrotic drug sera. The contents of astragaloside Ⅳ in fibrotic drug sera were higher than normal drug sera (0.1140 vs 0.0851,P<0.05 ). Astragaloside Ⅳ had inhibitory effect on proliferation of HSCs and the synthesis of collagen I in HSCs. Conclusion: HPLC is suitable for the determination of bioactive components in drugs (especially Traditional Chinese Medicine, TCM) in vitro and after metabolized in vivo under different conditions. The results show that astragaloside Ⅳ could be found in both raw astragalus mongholicus and its corresponding drug sera. The different conditions for metabolism might affect the contents of astragaloside Ⅳ. The concentration of Astragaloside Ⅳ in fibrotic drug sera is higher than that in normal sera, which may be caused by different “first pass effect” under different liver conditions and the different analytic metabolism. This phenomenon is in accordance with the different curative effects of different patients accepting TCM in clinic. Astragaloside Ⅳ, which could inhibit the proliferation and activation of HSCs, might be a working component in astragalus mongholicus.
Key words:Astragaloside Ⅳ; Seropharmacological method; High performance liquid chromatography (HPLC); Liver fibrosis; Hepatic stellate cells (HSCs), 百拇医药(吕涛 姚希贤 孙泽明)
关键词：黄芪甲苷；血清药理学；高效液相色谱法；肝纤维化；肝星状细胞
, http://www.100md.com
中图分类号：R285.5文献标识码：A文章编号：1673-7717(2011)12-2770-04
Content Assaying of Astragaloside IV and Its Effects on Proliferation and Activation of
HSCs in Liver Fibrosis
LU Tao1, YAO Xixian2, SUN Zeming2
(1. Dapartment of Gastroenterology, First People's Hospital of Hangzhou, Hangzhou 310006, Zhejiang, China;
2. Hebei Institute of Gastroenterology, Shijiazhuang 050000, Hebei, China)
, 百拇医药
________________________________________
收稿日期：2011-07-10
作者简介：吕涛(1976-)，女，河北石家庄人，医师，博士，研究方向：中药防治肝纤维化作用机制及成分研究。Abstract:Objective: To detect the contents of astragaloside Ⅳ in raw astragalus mongholicus, and the corresponding drug sera (normal and fibrotic drug sera) by HPLC. To compare the astragaloside Ⅳ in astragalus mongholicus. To study the effects of astragaloside Ⅳon proliferation and activation of HSCs in hepatic fibrogenesis. Methods: Inducing liver fibrosis in rats by 40% CCl4, SC, for 9 weeks. Astragalus mongholicus. solutions were given to the normal and fibrotic rats, respectively for 5days. Normal and fibrotic drug sera from rats were extracted. The separation of astragaloside Ⅳwas performed by HPLC. The effects of astragaloside Ⅳ on the proliferation of HSCs were measured by the method of CCK-8 (Cell Counting Kit-8). The effects on synthesis of collagen I were assessed by 3H-proline incorporation assays. Results: Astragaloside Ⅳ could be found both in normal and fibrotic drug sera. The contents of astragaloside Ⅳ in fibrotic drug sera were higher than normal drug sera (0.1140 vs 0.0851,P<0.05 ). Astragaloside Ⅳ had inhibitory effect on proliferation of HSCs and the synthesis of collagen I in HSCs. Conclusion: HPLC is suitable for the determination of bioactive components in drugs (especially Traditional Chinese Medicine, TCM) in vitro and after metabolized in vivo under different conditions. The results show that astragaloside Ⅳ could be found in both raw astragalus mongholicus and its corresponding drug sera. The different conditions for metabolism might affect the contents of astragaloside Ⅳ. The concentration of Astragaloside Ⅳ in fibrotic drug sera is higher than that in normal sera, which may be caused by different “first pass effect” under different liver conditions and the different analytic metabolism. This phenomenon is in accordance with the different curative effects of different patients accepting TCM in clinic. Astragaloside Ⅳ, which could inhibit the proliferation and activation of HSCs, might be a working component in astragalus mongholicus.
Key words:Astragaloside Ⅳ; Seropharmacological method; High performance liquid chromatography (HPLC); Liver fibrosis; Hepatic stellate cells (HSCs), 百拇医药(吕涛 姚希贤 孙泽明)