基于液质联用技术及抗炎与抗氧化活性研究夏枯草茎叶和果穗的替代性

Substitutability of Prunella vulgaris L Stem Leaf and Ear Based on HPLC-ESI-MSnAnalysis and Anti-Inflammatory and Antioxidant Activity

目的比较与分析夏枯草茎叶与果穗化学组分与药效的差异,为夏枯草茎叶能否替代果穗入药或共同入药提供有效的实验依据。方法采用液质联用分析法,对不同产地夏枯草茎叶与果穗水提物进行成分分析;通过角叉菜胶致大鼠足跖肿胀、二甲苯致小鼠耳郭肿胀实验模型,并采用酶联免疫吸附法测定大鼠足跖肿胀实验大鼠血浆中肿瘤坏死因子-α(TNF-α)浓度;观察比较夏枯草茎叶与果穗水提物的抗炎效果,运用快速ABTS检测法比较夏枯草茎叶与果穗水提物的总抗氧化能力。结果本实验所用不同产地夏枯草茎叶与果穗在化学组分上无显著性差异,主要含有三萜、黄酮、酚酸等物质;夏枯草果穗组在1 h开始显著抑制大鼠足跖肿胀,而夏枯草茎叶组在2 h才对大鼠足跖肿胀产生显著性抑制效果,并且均能显著降低肿瘤坏死因子-α的含量;夏枯草茎叶与果穗水提物均能显著减轻小鼠耳郭肿胀;夏枯草茎叶与果穗水提物均表现出较强的抗氧化活性,最高接近Trolox标准溶液的1.5倍。结论本实验样品夏枯草茎叶与果穗在化学组分上基本相似,仅含量有差异;夏枯草茎叶与果穗水提物均有一定的抗炎作用,且茎叶抗氧化活性较果穗强,因此茎叶替代果穗或共同入药具有可能性。

OBJECTIVE To compare the chemical composition and therapeutic effect between Prunella vulgaris L stem leaf and ear,thus to provide evidence for judging whether the stem leaf of Prunella vulgaris L can substitute the ear as an herbal medicine.METHODS Aqueous extracts of the stem leaf and ear of Prunella vulgaris L from different producing areas were analyzed with HPLCESI-MSn. The anti-inflammatory effects were observed by inflammatory models of ear edema induced by dimethylbenzene in mice and hind paw edema induced by carrageenan in rats. Enzyme-linked immunosorbent assay（ ELISA） was used to detect the plasma level of TNF-α and antioxidant activities were detected by ABTS method. RESULTS Prunella vulgaris L stem leaf and ear were not significantly different in chemical composition,both of which contained mainly triterpenoids,flavonoids,phenolic acids and other substances. Compared with the model group,Prunella vulgaris L ear significantly reduced the hind paw edema in rats induced by carrageenan from 1 h after oral administration（ P〈0. 05）,while the onset time of stem leaf was later than 1 h. Both groups could significantly reduce the ear edema in mice induced by dimethylbenzene（ P〈0. 01）. The TNF-α levels in the Prunella vulgaris L stem leaf and ear groups [（ 24. 16 ± 1. 24） and（ 24. 33 ± 2. 36） ng·mL^-1]were lower than that in the model group [（ 31. 34 ± 1. 94） ng·mL^-1]（ P〈0. 01）. Prunella vulgaris L stem leaf and ear groups showed strong antioxidant activities in the ABTS^· ＋scavenging test.CONCLUSION The contents of the main constituents in Prunella vulgaris L stem leaf and ear have significance differences. The results of animal tests indicate that the aqueous extracts of Prunella vulgaris L stem leaf and ear have significant anti-inflammatory and antioxidant effects.