桂枝附子含药血清对类风湿关节炎模型的影响
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摘要
目的通过液质联用(LC-MS)技术分析桂枝附子温通经脉配伍灌胃后胶原关节炎(CIA)大鼠的血清移行成分,考察桂枝附子含药血清对类风湿关节炎模型的影响。方法选取雄性Wistar大鼠25只随机取6只分为正常组、桂枝附子组,每组3只,其余大鼠采用Ⅱ型胶原造模。将造模成功的6只CIA大鼠随机平均分为模型组、造模桂枝附子组,每组3只。予20倍给药剂量的桂枝附子配伍灌胃给药,正常组和模型组大鼠予生理盐水灌胃,60 min后经肝门静脉取血,采用甲醇沉淀法处理血清,予LC-MS技术对其进行分析。通过将含药血清与相同条件下测定的模型组血清、正常组血清、供试品以及对照品图谱比较,分析大鼠给药后血清中的移行成分。结果桂枝附子组含药血清中产生了10个移行成分,均为代谢产物;造模桂枝附子组含药血清产生了25个移行成分,其中8个为原形成分,17个为代谢产物。结论造模后大鼠的含药血清中移行成分明显增加,成分浓度也较高。由此可见,CIA大鼠病理模型选择性的吸收了桂枝附子含药学清中的成分,从基础层面探讨中医"有是病用是药"理论。
Objective:To analyze the serum migration components of rats with collagen-induced arthritis(CIA)after administration of Guizhi Fuzi warming the meridians compatibility by liquid chromatography-mass spectrometry(LC-MS),and to study the effect of serum-containing components of Guizhi Fuzi on rheumatoid arthritis model. Methods:6 rats were randomly selected from 25 male Wistar rats and divided into a normal group and a Guizhi Fuzi group,3 rats each group. The other rats were modeled with type II collagen. 6 CIA rats with successful modeling were randomly divided into a model group and a model of Guizhi Fuzi group,3 rats each group. The rats in the Guizhi Fuzi group were administered with 20-time dose of Guizhi Fuzi and the rats in the normal group and the model group were given normal saline. After 60 minutes,the blood was taken from the hepatic portal vein. The serum was treated by methanol precipitation and analyzed by LC-MS. The migration components in the serum after administration of the rats were analyzed by comparing the drug-containing serum with the model group serum,and the serum of the normal group,the test sample,and the reference spectrum were measured under the same conditions. Results:10 transitional components were produced in the serum of Guizhi Fuzi group,all of which were metabolites. 25 migration components were produced in the modeling Guizhi Fuzi group,of which 8 were original components and 17 were metabolites. Conclusion:The migration component of the drug-containing serum of the rats after modeling is significantly increased,and the concentration of the components is also high. It can be seen that the pathological model of CIA rats selectively absorbs the components of collateral aconite containing drug serum,and basically discusses the theory of "to have disease or to use drug".
Objective:To analyze the serum migration components of rats with collagen-induced arthritis(CIA)after administration of Guizhi Fuzi warming the meridians compatibility by liquid chromatography-mass spectrometry(LC-MS),and to study the effect of serum-containing components of Guizhi Fuzi on rheumatoid arthritis model. Methods:6 rats were randomly selected from 25 male Wistar rats and divided into a normal group and a Guizhi Fuzi group,3 rats each group. The other rats were modeled with type II collagen. 6 CIA rats with successful modeling were randomly divided into a model group and a model of Guizhi Fuzi group,3 rats each group. The rats in the Guizhi Fuzi group were administered with 20-time dose of Guizhi Fuzi and the rats in the normal group and the model group were given normal saline. After 60 minutes,the blood was taken from the hepatic portal vein. The serum was treated by methanol precipitation and analyzed by LC-MS. The migration components in the serum after administration of the rats were analyzed by comparing the drug-containing serum with the model group serum,and the serum of the normal group,the test sample,and the reference spectrum were measured under the same conditions. Results:10 transitional components were produced in the serum of Guizhi Fuzi group,all of which were metabolites. 25 migration components were produced in the modeling Guizhi Fuzi group,of which 8 were original components and 17 were metabolites. Conclusion:The migration component of the drug-containing serum of the rats after modeling is significantly increased,and the concentration of the components is also high. It can be seen that the pathological model of CIA rats selectively absorbs the components of collateral aconite containing drug serum,and basically discusses the theory of "to have disease or to use drug".
引文
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[11]吕涛,姚希贤,孙泽明.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[J].中华中医药学刊,2011,12:2770-2773.
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[2]刘卓,晏星,徐春霞,等.附子总生物碱的提取工艺研究[J].安徽农业科学,2012,9:5199-5201.
[3]高晓霞.桂枝茯苓胶囊的代谢组学与药代动力学研究[D].沈阳药科大学,2009.
[4]王喜军.中药血清药物化学[M].北京:科学出版社,2010:285.
[5]张庆祥.辨病论治与辨证论治关系探析[J].辽宁中医杂志,2005,2:107-108.
[6]Yang W Z,Ye M,Qiao X,et al.A strategy for efficient discovery of new natural co-mpounds by integrating orthogonal column chromatography and liquid chromatography/mass spectrometry analysis:its application in Panax ginseng,Panax quinquefoliumand Panax noto-ginseng to characterize 437 potential new ginsenosides[J].Anal Chim Acta,2012,739:56.
[7]雷志丹,雷志钧,夏新华.川芎血清药物化学的初步研究[J].中国实验方剂学杂志,2011,12:96-98.
[8]张星华,杜小正,王金海,等.类风湿关节炎动物模型在中医药研究中的进展[J].亚太传统医药,2015,13:47-50.
[9]赵宏艳,王燕,肖诚,等.不同品系、不同性别对大鼠CIA发病情况的比较研究[J].中国中医基础医学杂志,2010,09:761-764.
[10]殷飞,姚树坤,吴新满,等.清肝化瘀方药含药血清对肝癌细胞SMMC-7721增殖和凋亡的影响[J].中药药理与临床,2004,20(3):30.
[11]吕涛,姚希贤,孙泽明.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[J].中华中医药学刊,2011,12:2770-2773.
[12]韩笑,刘建勋,林成仁.血清药理学关键问题的存疑与讨论[J].中药药理与临床,2009,25(2):122-124.