摘要
目的基于两种不同取血部位制备桂枝汤含药血清,探讨其抗炎作用差异。方法取雄性SD大鼠按10倍临床剂量给药(桂枝汤汤剂生药1 g/mL),取门静脉血、外周血含药血清,采用快速液相-三重四级杆质谱联用仪(rapid resolution liquid chromatography-triple quadrupole mass spectrometry,RRLC-QQQ-MS/MS),建立桂枝汤汤剂、含药血清中8种化学成分含量测定方法,并对含量进行测定。取对数生长期小鼠脑微血管内皮细胞(brain-derived endothelial cells.3,bEnd.3)模型,用白细胞介素(interleukin,IL)-1β刺激,建立细胞炎症模型,使用bEnd.3细胞作为正常组。除正常组、模型组外,设置桂枝汤汤剂高、中、低剂量组(分别含5%、2.5%、1.25%桂枝汤汤剂),门静脉含药血清高、中、低剂量组(分别含20%、10%、5%门静脉含药血清)和外周血含药血清高、中、低剂量组(分别含20%、10%、5%外周血含药血清),ELISA法测定细胞外液中的前列腺素(prostaglandin,PG)E_(2)含量。结果含量测定结果显示汤剂中芍药苷、芍药内酯、甘草苷、甘草酸比例相对较高,血清中肉桂酸、6-姜酚含量相对较高,较汤剂中新出现甘草次酸等代谢物。门静脉血清中以上8种化合物含量均高于外周血血清。5%桂枝汤汤剂中甘草酸含量为798.13 ng/mL,20%门静脉含药血清中甘草酸含量为316.22 ng/mL,20%外周血中甘草酸含量为481.17 ng/mL,药物中成分的含量在同一数量级。与模型组比较,将高、中、低剂量桂枝汤汤剂添加到bEnd.3细胞培养体系中,高、中、低剂量桂枝汤组细胞外液中PGE_(2)的水平均明显高于模型组(P<0.05),而门静脉或外周血含药血清组细胞外液中PGE_(2)的水平均明显低于模型组(P<0.05),且门静脉血清的药效作用显著强于相同浓度的外周血血清(P<0.05)。结论桂枝汤汤剂经胃肠道给药后,门静脉含药血清相较外周血含药血清显示出更高的成分含量与抗炎作用。这一发现提示采用门静脉含药血清而非传统的外周血含药血清在中药方剂研究中的重要性。
Objective To investigate the differences in anti-inflammatory effects between drug-containing serum prepared from two different blood collection sites using Guizhi decoction.Methods Male SD rats were administered Guizhi decoction at a dosage of 1 g/mL,ten times the clinical dose.Drug-containing serum was collected from both the portal vein and peripheral blood.RRLC-QQQ-MS/MS method was developed to quantify eight chemical constituents in both the Guizhi decoction and the drug-containing serum.Mouse brain-derived endothelial cells(bEnd.3)in the logarithmic growth phase were utilized to establish an inflammation model stimulated by IL-1β,with bEnd.3 cells also serving as the control group.Apart from the control and model groups,there were high,medium,and low dosage groups for Guizhi decoction(5%,2.5%,and 1.25%of the decoction respectively),as well as high,medium,and low dosage groups for drug-containing serum from the portal vein and peripheral blood(20%,10%,and 5%of serum respectively).The levels of PGE_(2)in the extracellular fluid were measured by ELISA.Results The analysis revealed that the decoction had relatively high concentrations of paeoniflorin,paeonol,glycyrrhizin,and glycyrrhetinic acid,while the serum was rich in cinnamic acid and 6-gingerol,along with new metabolites like glycyrrhetinic acid monoglucuronide.The content of these compounds was higher in the portal vein drug-containing serum compared to the peripheral blood serum.The concentration of glycyrrhetinic acid in 5%Guizhi decoction was 798.13 ng/mL,in 20%portal vein drug-containing serum was 316.22 ng/mL,and in 20%peripheral blood serum was 481.17 ng/mL,indicating similar levels of constituents in the drug across these mediums.When compared with the model group,the PGE_(2)levels in the extracellular fluid of bEnd.3 cells treated with varying doses of Guizhi decoction were significantly higher(P<0.05),whereas those in groups treated with drug-containing serum from both the portal vein and peripheral blood were significantly lower(P<0.05).Furthermore,the portal vein serum exhibited a markedly stronger pharmacological effect than the peripheral blood serum at equivalent concentrations(P<0.05).Conclusion Post-oral administration of Guizhi decoction,the drug-containing serum from the portal vein exhibited higher constituent levels and more potent anti-inflammatory effects compared to the peripheral blood serum.This finding highlights the significance of using portal vein drug-containing serum over traditional peripheral blood serum in research on Chinese herbal formulas.
作者
阎政燊
张立石
高焕
郭琴
宋剑南
高源
柏冬
YAN Zhengshen;ZHANG Lishi;GAO Huan;GUO Qin;SONG Jiannan;GAO Yuan;BAI Dong(Institute of Basic Theory for Chinese Medicine,China Academy of Chinese Medical Sciences,Beijing 100700,China;Shaanxi University of Chinese Medicine,Xianyang 712046,China)
出处
《中国中医基础医学杂志》
CAS
CSCD
2024年第5期828-835,共8页
JOURNAL OF BASIC CHINESE MEDICINE
基金
中国中医科学院科技创新工程项目(C12021A00108)
国家自然科学基金项目(81403282)
北京市自然科学基金项目(7133251)。
关键词
桂枝汤
门静脉含药血清
外周血含药血清
含量测定
药效实验
Guizhi decoction
Drug-containing serum of portal vein blood
Drug-containing serum of peripheral blood
Content determination
Pharmacodynamic experiment
