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金茵利胆胶囊的体外抗炎活性评价及其谱效关系
曹凡1,宋忠兴2,陈琳2*,唐志书2,3*,倪健4,张德柱5
0
(1. 陕西中医药大学药学院, 西安 712046;
2. 陕西省中药资源产业化协同创新中心, 秦药特色资源研究开发国家重点实验室(培育), 陕西省创新药物研究中心, 陕西中医药大学, 咸阳 712083;
3. 中国中医科学院中药资源中心, 北京 100700;
4. 北京中医药大学北京中医药研究院, 北京 100029;
5. 陕西盘龙药业集团股份有限公司, 西安 710025
*通信作者)
摘要:
目的 评价金茵利胆胶囊的体外抗炎活性,并研究其抗炎谱效关系。方法 建立脂多糖(LPS)诱导的小鼠单核巨噬细胞(RAW264.7细胞)炎症模型,检测细胞培养上清液中一氧化氮(NO)、TNF-α和IL-6等的含量以评价金茵利胆胶囊的抗炎活性。采用均匀设计法制备金茵利胆胶囊组成饮片的不同配伍样本,采用HPLC建立其指纹图谱,并进行抗炎活性测定;以细胞培养上清液中NO、TNF-α和IL-6的抑制率为药效指标,与共有峰峰面积经灰色关联分析构建抗炎谱效关系。结果 金茵利胆胶囊提取物(0.25~1.00 mg/mL)能抑制LPS诱导的RAW264.7细胞分泌NO、TNF-α和IL-6。经对照品比对指认出其中7个色谱峰,分别为新绿原酸、绿原酸、隐绿原酸、1,3-二咖啡酰奎宁酸、对羟基苯乙酮、柚皮苷、新橙皮苷。抗炎谱效关系分析结果表明,23个共有峰均具有一定的抗炎贡献度,其中峰21(柚皮苷)、峰23(新橙皮苷)与3个细胞炎症指标(NO、TNF-α、IL-6)的关联度均>0.74。结论 金茵利胆胶囊发挥抗炎作用是多种成分共同作用的结果。本研究结果可为金茵利胆胶囊抗炎药效物质基础及其质量控制提升提供参考。
关键词:  金茵利胆胶囊  单核巨噬细胞  抗炎活性  均匀设计  谱效关系
DOI:10.16781/j.CN31-2187/R.20220091
投稿时间:2022-01-24修订日期:2022-04-29
基金项目:中药大品种品牌价值提升示范研究项目(202190025),国家现代农业产业技术体系资助项目(CARS-21),咸阳市2020年度重大科技专项(2020K01-20),国家自然科学基金青年科学基金(81904047),陕西省千人计划区域青年人才项目(2018).
Evaluation for in vitro anti-inflammatory activity of Jinyin Lidan capsule and its spectrum-effect relation
CAO Fan1,SONG Zhong-xing2,CHEN Lin2*,TANG Zhi-shu2,3*,NI Jian4,ZHANG De-zhu5
(1. College of Pharmacy, Shaanxi University of Chinese Medicine, Xi'an 712046, Shaanxi, China;
2. Shaanxi Collaborative Innovation Center of Chinese Medicine Resources Industrialization, State Key Laboratory of Research and Development of Characteristic Qin Medicine Resources (Cultivation), Shaanxi Innovative Drug Research Center, Shaanxi University of Chinese Medicine, Xianyang 712083, Shaanxi, China;
3. Chinese Medicine Resource Center, China Academy of Chinese Medical Sciences, Beijing 100700, China;
4. Beijing Academy of Chinese Medicine, Beijing University of Chinese Medicine, Beijing 100029, China;
5. Shaanxi Panlong Pharmaceutical Group Co. Ltd., Xi'an 710025, Shaanxi, China
*Corresponding authors)
Abstract:
Objective To evaluate the in vitro anti-inflammatory activity of Jinyin Lidan capsule and study its anti-inflammatory spectrum-effect relation. Methods The lipopolysaccharide (LPS)-induced inflammation model of mouse mononuclear macrophages (RAW264.7 cells) was established, and the contents of nitric oxide (NO), tumor necrosis factor α (TNF-α) and interleukin 6 (IL-6) in the supernatant of RAW264.7 cells were determined to evaluate the anti-inflammatory activity of Jinyin Lidan capsule. Different compatible samples of Jinyin Lidan capsules were prepared by uniform distribution design method, their fingerprints were established by high-performance liquid chromatography (HPLC), and their anti-inflammatory activity was determined. The inhibitory rates of NO, TNF-α and IL-6 in the supernatant were used as pharmacodynamic indexes, and the correlation between the anti-inflammatory spectrum and the common peak area was established by grey correlation analysis. Results The Jinyin Lidan capsule extract (0.25-1.00 mg/mL) reduced the secretion of NO, TNF-α and IL-6 in LPS-induced inflammatory RAW264.7 cells. Seven chromatographic peaks were identified by reference substance comparison, and they were neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, 1,3-dicaffeoylquinic acid, p-hydroxyacetophenone, naringin, and neohesperidin. The analysis results of anti-inflammatory spectrum-effect relation showed that 23 common peaks had a certain degree of anti-inflammatory contribution, and the correlation degree of peak 21 (naringin) and peak 23 (neohesperidin) with the 3 cellular inflammatory indicators (NO, TNF-α, and IL-6) was greater than 0.74. Conclusion The anti-inflammatory effect of Jinyin Lidan capsule is the result of the joint action of various components, and the results of our study can provide reference for the improvement of the substance basis and quality control of Jinyin Lidan capsule.
Key words:  Jinyin Lidan capsule  mononuclear phagocyte  anti-inflammatory activity  design by uniform distribution  spectrum-effect relation