National Omics Centre, National Science and Technology Development Agency (NSTDA), Pathum Thani, Thailand.
Department of Biochemistry, Faculty of Science, Kasetsart University, Bangkok, Thailand.
Phytochem Anal. 2022 Oct;33(7):1086-1098. doi: 10.1002/pca.3161. Epub 2022 Jul 5.
Bua Bok or Centella asiatica (CA) is an Asian vegetable with anti-inflammatory benefits. Asiaticoside, asiatic acid, madecassoside and madecassic have been characterised as major active ingredients with a wide range of pharmacological advantages. In manufacturing processes, high-performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LCMS) are used to routinely determine the active compounds in raw materials.
This research aims to explore anti-inflammatory properties, characterise metabolites and observe the biochemical changes of the inflammatory induced macrophages after pretreatment with the potential extracted fractions.
Bua Bok leaf extracts were prepared. Macrophages were pretreated with non-toxic fractions to determine the anti-inflammatory action. Tentative metabolites of effective fractions were identified by LC-MS. Synchrotron fourier-transform infrared (S-FTIR) microspectroscopy was utilised to observe the biochemical change of the lipopolysaccharide (LPS)-induced cells after pretreatment with potential fractions.
Fractions of ethyl acetate, 30% and 100% ethanol highly increased the nitrile scavenging and suppressed the function of phospholipase A . Fractions of 70% and 100% ethanol strongly decreased nitric oxide production. The comparison of 39 chemical compounds was presented. The change of proteins was improved after pretreatment of macrophages with fraction 70% ethanol. Fraction of 100% ethanol revealed the lipid accumulation was lower than 70% ethanol and diclofenac.
While the anti-inflammatory actions of 70% and 100% ethanol were similar. S-FTIR expressed they inhibited inflammatory response with the distinct features of biomolecules. The S-FTIR, LC-MS and biological assay confidently provided the efficient strategies to inform the advantage of herbal extract on cellular organisation instead of a single compound.
积雪草（CA）又名布哇或积雪草，是一种具有抗炎功效的亚洲蔬菜。已鉴定出积雪草酸、积雪草苷、羟基积雪草苷和羟基积雪草酸为主要活性成分，具有广泛的药理优势。在制造过程中，高效液相色谱（HPLC）和液相色谱-质谱联用（LCMS）常用于常规测定原料中的活性化合物。
本研究旨在探讨积雪草提取物的抗炎特性，对代谢产物进行表征，并观察经潜在提取部位预处理后诱导的巨噬细胞的生化变化。
制备积雪草叶提取物。用无毒部位预处理巨噬细胞，以确定抗炎作用。通过 LC-MS 鉴定有效部位的推测代谢产物。利用同步辐射傅里叶变换红外（S-FTIR）微光谱法观察潜在部位预处理后脂多糖（LPS）诱导细胞的生化变化。
乙酸乙酯、30%和 100%乙醇部位高度增加了腈的清除，并抑制了磷脂酶 A 的功能。70%和 100%乙醇部位强烈降低了一氧化氮的产生。比较了 39 种化合物。用 70%乙醇预处理巨噬细胞后，蛋白质的变化得到改善。与 70%乙醇和双氯芬酸相比，100%乙醇部位显示出较低的脂质积累。
虽然 70%和 100%乙醇的抗炎作用相似，但 S-FTIR 表明它们通过生物分子的独特特征抑制了炎症反应。S-FTIR、LC-MS 和生物测定为告知草药提取物对细胞组织的优势提供了有效的策略，而不是单一化合物。
