金银花与山银花抗流感病毒功效差异的关键药效组分筛选及其作用机制研究

The phenomenon of the same effect medicinal materials of multiple primaries is one of TCM features, at the same time also is the main factor that leads to effect not stable. It is necessary than the main effect different is systematically assessed based on key efficacy components screening in medicinal materials of multiple primaries. Lonicerae Japonicae Flos and L. Flos are typical representatives for this kind of medicinal materials. However, the efficacy 'heat-clearing and detoxifying, evacuation wind heat' is uniformly described in L. Japonicae Flos and L. Flos in Chinese Pharmacopoeia. We have not still known whether there were the effect differences in both medicinal materials up to now, thus huge troubles have been hidden when facing with a major epidemic situation. The further studies will be carried out based on early chemistry and pharmacodynamics works in L. Japonicae Flos and L. Flos. The methods of analytical chemistry and pharmacodynamics are used synthetically in this study. Key efficacy components and their the best proportion of efficacy differences are confirmed by establishing the spectrum-effect relationship between chemical information and anti influenza virus efficacy information in vitro test. Action mechanisms of efficacy differences are expounded by investigating death protection, lung tissue inflammation inhibition and pathology, lung tissue cell apoptosis, immune function on mice model in vivo test. Lastly, anti influenza virus effects between L.Japonicae Flos and L.Flos are synthetically evaluated based on the results. The results suggest important theoretical values and practical significances are put forward to guide the clinical medication ensure curative effect, develop healthily industry of L.Japonicae Flos and L.Flos, and enrich basic theory of medicinal materials of multiple primaries quality research.

"一药多基原"现象是中药一大特色，同时也是导致功效不稳定的重要因素。因此，对于多基原药材有必要在筛选其关键药效成分的基础上，对其主要功效差异进行系统评价。金银花与山银花为这类药材的典型代表，两者在药典中的功效描述均为"清热解毒，疏散风热"，但两者间的功效差异尚不明确，这在面临重大疫情时潜伏着巨大隐患。 本研究是申请者前期对金银花与山银花化学及药效学研究工作的深入。拟综合运用分析化学和药理学的手段，建立化学信息与体外抗流感病毒药效信息的谱效关系，筛选出导致两者功效差异的关键药效组分及其黄金比例；从对流感病毒模型小鼠的死亡保护作用、肺组织炎症抑制作用、肺组织细胞凋亡、机体免疫功能的影响角度，阐明功效差异的作用机制；最后，综合评价金银花与山银花抗流感功效。研究结果对于指导临床用药确保疗效，对于金银花与山银花产业的健康发展，丰富多基原药材品质研究的基础理论，都具有重要的理论价值和实践意义。

金银花是传统中药材，也是重要的药食同源药材，近年来在抗击非典、甲型流感病毒中起到重要作用。金银花的近缘种是山银花，由于某些原因，两者长期混用，而《药典》中两者的性味与归经、功能与主治、用法与用量完全相同，它们之间的药效到底有何差异呢？ .本项目重点研究了金银花与山银花在抗流感病毒关键药效组分组成及其作用机制。取得了如下重要结果：①化学成分上的差异：山银花中总有机酸、总黄酮、总皂苷含量比金银花多，而金银花较山银花含有更丰富的环烯醚萜类物质。②抗流感病毒作用差异：金银花和山银花水提液对H1N1流感病毒致细胞病变皆有一定的抑制作用，以治疗指数（TI）为评价标准，药效由高到低依次排列分别为：红腺忍冬的TI是17.71±9.80，金银花的TI是13.17±4.60，灰毡毛忍冬的TI是10.60±3.75，黄褐毛忍冬的TI是10.50±2.71，华南忍冬的TI是6.84±1.74。③谱效关系研究中发现：金银花有8个关键成分，山银花中的灰毡毛忍冬有5个关键成分，华南忍冬有5个关键成分，黄褐毛忍冬有3个关键成分，红腺忍冬有2个关键成分。本研究结果体现了中药多组分的协同作用，通过标准品比对发现马钱苷，咖啡酸，阿魏酸，木犀草苷，异绿原酸A，金丝桃苷，绿原酸在抗病毒效果中起到重要作用。④抗流感病毒功效差异的作用机制：在抗流感病毒H1N1方面，就肺部炎症抑制作用，肺指数，肺组织病理切片及对肺炎小鼠的免疫作用来看，金银花与山银花都具有一定的治疗效果。和病毒组相比，红腺忍冬在较大程度上降低流感病毒肺炎小鼠肺组织细胞凋亡率；红腺忍冬组CD4+T细胞百分比有显著性差异（P<0.01），IL-1β含量显著性降低（P<0.05），流感病毒肺炎小鼠IgG水平有显著性提高（P<0.05），其中来源于红腺忍冬的山银花具有较好的治疗效果，与金银花效果相当。.本研究结果对于指导临床用药确保疗效，对于金银花与山银花产业的健康发展，丰富“多基原”药材品质研究的基础理论，都具有重要的理论价值和实践意义。