Abstract
The metabolomic screening of potential anti-inflammatory compounds in the leaves of Actinidia arguta was performed by using LC-MS/MS. Ethanol extracts were prepared, and the anti-inflammatory effects were investigated based on nitric oxide (NO) synthesis and inducible nitric oxide synthase expression in lipopolysaccharide-induced RAW 264.7 macrophages. The 75% ethanol extract showed the highest inhibitory effect on nitric oxide (NO) production, and it was further separated by in vitro bioassay-guided fractionation using preparative LC with reversed-phase column separation. Through multiple steps of fractionation, sub-fraction 1-3 was finally purified, and caffeic acid derivatives, such as caffeoylthreonic acid and danshensu (salvianic acid A), were successfully identified as key anti-inflammatory compounds by LC-MS/MS and metabolomics analyses. This is the first study identifying anti-inflammatory compounds in A. arguta (Actinidia arguta) leaves through bioassay-guided fractionation and metabolomics screening. Results of this study would be useful for the application of A. arguta leaves as a nutraceutical.
Highlights
Actinidia arguta (Actinidiaceae) is native to China, Korea, Siberia, and Japan
The 75% ethanol extract exhibited the highest extraction yield (20.15 ± 0.85%), whereas the yields were 10.78 ± 2.15%, 13.68 ± 1.09%, 15.89 ± 1.18%, and 4.71 ± 0.15% after extraction with distilled water, 25% ethanol, All extracts and their fractions inhibited LPS-induced nitric oxide (NO) production in a dose-dependent manner
The 75% ethanol extracts were partially purified by preparative LC (prep-LC)
Summary
In Korea, fruits, stems, roots, and leaves of A. arguta (Actinidia arguta) are traditionally used as medicines. The leaves are used as a folk medicine to cure intestinal catarrh, stomach cancer, and acute gastritis in Korea [4]. Phytochemicals in A. arguta leaves, such as flavonoids [7], lignin, and phenolic acid [8], have been reported as key anti-diabetic compounds. Webby et al investigated flavonoids in the leaf of Actinidia, but the study was only focused on several compounds, such as quercetin, kaempferol, and myricetin, with their glycosides [7]. Kwak and Lee [9] reported on the antioxidant and anti-inflammatory effects of ethanol extracts from A. arguta leaves, the systematic analysis of its key compounds has not been conducted
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