Identification of bioactive phenolics from Porana sinensis Hemsl. stem by UPLC-QTOF-MS/MS and the confirmation of anti-inflammatory indicators using LPS-induced RAW264.7 cells
- 89 Downloads
To characterise bioactive phenolics and confirm anti-inflammatory indicators in Porana sinensis stem, 23 phenolics were identified by UPLC-QTOF-MS/MS from crude extract (CE) prepared optimally with 80% methanol. Further fractionalisation using D101 macroporous resin resulted in predominant enrichment of total phenols and flavonoids into Fr.II. Correspondingly, the bioactive components-enriched Fr.II exhibited the lowest IC50 for scavenging DPPH and ABTS and the highest oxygen radical absorbance capacity or ORAC followed by Fractions Fr.I + Fr.II, CE and Fr.I, implying that certain phenolics possessing lower antioxidant activity completely remained in CE. Anti-inflammatory tests with LPS-stimulated RAW264.7 cells showed that CE possessed the highest inhibition of NO-production followed by Fr.II and Fr.I, meaning that CE might contain compounds that expressed higher anti-inflammatory but lower antioxidant activities or possessed synergistic interactions but were not fractionated together. Quantitative determination of nine major phenolics revealed that caffeic acid and 3-, 4- and 5-caffeoylquinic acids were concentrated into Fr.I, whereas scopolin, scopoletin and 3,5-, 3,4- and 4,5-dicaffeoylquinic acids were enriched into Fr.II. Further experiments with three selected major phenolics reduced the proposed synergistic interactions. Anti-inflammatory tests of the nine major phenolics evidenced that caffeic acid and the six caffeoylquinic acids produced higher, and the three dicaffeoylquinic acids at 140 μΜ showed even more significant activities in suppressing NO-production and mRNA expression of iNOS, TNF-α, COX-2, and IL-6, suggesting that these three dicaffeoylquinic acids could be indicators of the anti-inflammatory potential of P. sinensis stem. These findings provided novel insights for potential use of P. sinensis or liana, as an important source of natural antioxidants, against inflammation.
KeywordsPorana sinensis UPLC-QTOF-MS/MS D101 macroporous resin Phenolic antioxidant LPS-stimulated RAW264.7 cells Anti-inflammatory activity
This research was financially supported by the China Special Fund for Forestry Research in the Public Interest (Grant No. 201504606).
QX, LS, and YL designed the study, performed the research and drafted the manuscript. PY, KL, HF, LY, and XC participated in the experiments. YL provided the facilities and reviewed the manuscript. All authors read and approved the final manuscript.
Compliance with ethical standards
Conflict of interest
The authors declared that there were no conflicts of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
- Apel K, Hirt H (2004) Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu Rev Plant Biol 55:373–399. https://doi.org/10.1146/annurev.arplant.55.031903.141701 CrossRefGoogle Scholar
- Cai H, Harrison DG (2000) Endothelial dysfunction in cardiovascular diseases: the role of oxidant stress. Circ Res 87:840–844. http://circres.ahajournals.org/content/87/10/840
- Chen Z, Liao L, Yang Y, Zhang Z, Wang Z (2015) Different fingerprinting strategies to differentiate Porana sinensis and plants of Erycibe by high-performance liquid chromatography with diode array detection, ultra high performance liquid chromatography with tandem quadrupole mass spectrometry, and chemometrics. J Sep Sci 38:231–238. https://doi.org/10.1002/jssc.201400861 CrossRefGoogle Scholar
- Chunying L, Wang X, Ge G, Lian W, Yongxin L, Chengjun S (2013) Identification and quantification of free, conjugate and total phenolic compounds in leaves of 20 sweet potato cultivars by HPLC-DAD and HPLC-ESI-MS/MS. Food Chem 141:2697–2706. https://doi.org/10.1016/j.foodchem.2013.05.009 CrossRefGoogle Scholar
- Cui YQ, Jia YJ, Zhang T, Zhang QB, Wang XM (2012) Fucoidan protects against lipopolysaccharide-induced rat neuronal damage and inhibits the production of proinflammatory mediators in primary microglia. CNS Neurosci Ther 18:827–833. https://doi.org/10.1111/j.1755-5949.2012.00372.x CrossRefGoogle Scholar
- Fang X, Wang J, Hao J, Li X, Guo N (2015) Simultaneous extraction, identification and quantification of phenolic compounds in Eclipta prostrata, using microwave-assisted extraction combined with HPLC-DAD-ESI-MS/MS. Food Chem 188:527–536. https://doi.org/10.1016/j.foodchem.2015.05.037 CrossRefGoogle Scholar
- Hsu YW, Chi KH, Huang WC, Lin WW (2001) Ceramide inhibits lipopolysaccharide-mediated nitric oxide synthase and cyclooxygenase-2 induction in macrophages: effects on protein kinases and transcription factors. J Immunol 166:5388–5397. https://doi.org/10.4049/jimmunol.166.9.5388 CrossRefGoogle Scholar
- Longo W, Panesar N, Mazuski J, Kaminski D (1998) Contribution of cyclooxygenase-1 and cyclooxygenase-2 to prostanoid formation by human enterocytes stimulated by calcium ionophore and inflammatory agents. Prostag Oth Lipid M 56:325–339. https://doi.org/10.1016/S0090-6980(98)00058-6 CrossRefGoogle Scholar
- Nascimento AM, Maria-Ferreira D, Dal Lin FT, Kimura A, de Santana-Filho AP, Mfp W (2017) Phytochemical analysis and anti-inflammatory evaluation of compounds from an aqueous extract of Croton cajucara Benth. J Pharmaceut Biomed 145:821–830. https://doi.org/10.1016/j.jpba.2017.07.032 CrossRefGoogle Scholar
- Wu L, Zhu E, Zhang Z, Wang Z (2005) Investigating original plant of Caulis Erycibes in Guangxi and identifying mainstream variety of Caulis Erycibes in market. Chin Trad Herb Drugs 36:1398–1400Google Scholar
- Zhang L, Ravipati AS, Koyyalamudi SR, Jeong SC, Reddy N, Smith PT, Bartlett J, Shanmugam K, Münch DG, Wu MJ (2011) Antioxidant and anti-inflammatory activities of selected medicinal plants containing phenolic and flavonoid compounds. J Agric Food Chem 59:12361–12367. https://doi.org/10.1021/jf203146e CrossRefGoogle Scholar
- Zhang JY, Zhang Q, Li N, Wang ZJ, Lu JQ, Qiao YJ (2013) Diagnostic fragment-ion-based and extension strategy coupled to DFIs intensity analysis for identification of chlorogenic acids isomers in Flos Lonicerae Japonicae by HPLC-ESI-MS(n). Talanta 104:1–9. https://doi.org/10.1016/j.talanta.2012.11.012 CrossRefGoogle Scholar