Bioassay-Guided Isolation of Iridoids and Phenylpropanoids from Aerial Parts of Lamium album and Their Anti-inflammatory Activity in Human Neutrophils[*]
received 28 January 2017
revised 13 March 2017
accepted 17 March 2017
03 April 2017 (eFirst)
In traditional medicine, flowers and aerial parts of Lamium album are assigned by their anti-inflammatory, antiseptic, and mucolytic activities, and are used in chronic bronchitis and pharyngitis as well as skin, vaginal, and cervical inflammation.
The aim of the present study was to compare effects of ethanolic extracts prepared from flowers and aerial parts of L. album on selected functions of human neutrophils, which are involved in an inflammatory response. In order to identify the compounds engaged in the anti-inflammatory activity of extracts, the bioassay-guided isolation of compounds was performed based on the inhibition of cytokine secretion by stimulated neutrophils.
The extracts were phytochemically characterized with the HPLC-DAD-MSn method. The inhibition of reactive oxygen species production by formyl-met-leu-phenylalanine- or phorbol 12-myristate 13-acetate-stimulated neutrophils was determined using luminol- or lucigenin-dependent chemiluminescence. The effect on myeloperoxidase secretion by neutrophils was established spectrophotometrically. The levels of cytokine (interleukin 8, TNF-α) production after extract treatment was measured by ELISA.
The most abundant constituents of extracts were phenylpropanoids, iridoids, flavonoids, and phenolic acids. Both extracts at concentrations of 25 and 100 µg/mL significantly inhibited reactive oxygen species production, and myeloperoxidase and interleukin 8 secretion. The phenylethanoid glycosides, such as lamiusides A, B, and C as well as 6″-O-β-D-glucopyranosylmartynoside, were isolated and identified. The cells treated with 6″-O-β-D-glucopyranosylmartynoside and lamiuside B produced 29.47 ± 7.11 % and 64.67 ± 5.25 % of interleukin 8, respectively, compared to non-treated control cells.
Our results support the traditional use of L. album and indicate it as a potential source of natural anti-inflammatory constituents, such as phenylpropanoids.
* Dedicated to Professor Dr. Max Wichtl in recognition of his outstanding contribution to pharmacognosy research.
- 1 Blaschek W. Wichtl – Teedrogen und Phytopharmaka, 6th edition. Stuttgart: Wissenschaftliche Verlagsgesellschaft; 2016: 365-366
- 2 DAC/NRF-Kommission, ABDA – Bundesvereinigung Deutscher Apothekerverbände. Deutscher Arzneimittel-Codex/Neus Rezeptur Formularium (DAC/NRF). Monographie T-017. Taubnesselkraut (Lamii albi herba). 5th edition Stuttgart: Govi-Verlag Pharmazeutischer Verlag GmbH; 2016
- 3 Yordanova ZP, Zhiponova MK, Iakimova ET, Dimitrova MA, Kapchina-Toteva VM. Revealing the reviving secret of the white dead nettle (Lamium album L.). Phytochem Rev 2014; 13: 375-389
- 4 Deschauer T. Illustrated phytotherapy. Book two. Maywood, Ill: Thos. Deschauer Publications; 1945. Available at http://www.swsbm.com/Ephemera/Deschauer-2.pdf Accessed January 20, 2016
- 5 Remigton JP, Woods HC. The dispensatory of the United States of America 1918, 20th ed. The Southwest School of Botanical Medicine. Available at. http://www.swsbm.com/Dispensatory/USD-1918-complete.pdf Accessed January 20, 2016
- 6 Trouillas P, Calliste CA, Allais DP, Simon A, Marfak A, Delage C, Duroux JL. Antioxidant, anti-inflammatory and antiproliferative properties of sixteen water plant extracts used in the Limousin countryside as herbal teas. Food Chem 2003; 80: 399-407
- 7 Paduch R, Wójciak-Kosior M, Matysik G. Investigation of biological activity of Lamii albi flos extracts. J Ethnopharmacol 2007; 110: 69-75
- 8 Todorov D, Dimitrova M, Shishkova K, Yordanova Z, Kapchina-Toteva V, Shishkov S. Comparative anti-herpes effects of the chloroform in vitro and in vivo extracts, derived from Lamium album L. Bulg J Agric Sci 2013; 19: 190-193
- 9 Zhang H, Rothwangl K, Mesecar AD, Sabahi A, Rong L, Fong HHS. Lamiridosins, hepatitis C virus entry inhibitors from Lamium album . J Nat Prod 2009; 72: 2158-2162
- 10 Alipieva K, Kokubun T, Taskova R, Evstatieva L, Handjieva N. LC-ESI-MS analysis of iridoid glucosides in Lamium species. Biochem Syst Ecol 2007; 35: 17-22
- 11 Damtoft S. Iridoid glucosides from Lamium album . Phytochemistry 1991; 31: 175-178
- 12 Budzianowski J, Skrzypczak L. Phenylpropanoid esters from Lamium album flowers. Phytochemistry 1995; 38: 997-1001
- 13 Pereira OR, Domingues MRM, Silva AMS, Cardoso SM. Phenolic constituents of Lamium album: focus on isoscutellarein derivatives. Food Res Int 2012; 48: 330-335
- 14 Wójciak-Kosior M, Krzaczek T, Matysik G, Skalska A. HPTLC-densitometric method of determination of oleanolic acid in the Lamii albi flos . J Sep Sci 2005; 28: 2139-2143
- 15 Damtoft S, Jensen SR. Hemialboside, a hemiterpene glucoside from Lamium album . Phytochemistry 1995; 39: 923-924
- 16 Savchenko T, Blackford M, Sarker SD, Dinan L. Phytoecdysteroids from Lamium spp: identification and distribution within plants. Biochem Syst Ecol 2001; 29: 891-900
- 17 Quade MJ, Roth JA. A rapid, direct assay to measure degranulation of bovine neutrophil primary granules. Vet Immunol Immunopathol 1997; 58: 239-248
- 18 Sato T, Hongu T, Sakamoto M, Funakoshi Y, Kanaho Y. Molecular mechanisms of N-formyl-methionyl-leucyl-phenylalanine-induced superoxide generation and degranulation in mouse neutrophils: phospholipase D is dispensable. Mol Cell Biol 2013; 33: 136-145
- 19 Jaken S. Protein kinase C isozymes and substrates. Curr Opin Cell Biol 1996; 8: 168-173
- 20 Pålsson-McDermott EM, OʼNeill LA. Signal transduction by the lipopolysaccharide receptor, Toll-like receptor-4. Immunology 2004; 113: 153-162
- 21 Arnhold J, Flemmig J. Human myeloperoxidase in innate and acquired immunity. Arch Biochem Biophys 2010; 500: 92-106
- 22 Di Paola R, Esposito E, Mazzon E, Riccardi L, Caminiti R, Dal Toso R, Pressi G, Cuzzocrea S. Teupolioside, a phenylpropanoid glycosides of Ajuga reptans, biotechnologically produced by IRBN22 plant cell line, exerts beneficial effects on a rodent model of colitis. Biochem Pharmacol 2009; 77: 845-857
- 23 Ito N, Nihei T, Kakuda R, Yaoita Y, Kikuchi M. Five new phenylethanoid glycosides from the whole plants of Lamium purpureum L. Chem Pharm Bull (Tokyo) 2006; 54: 1705-1708
- 24 Mittal M, Siddiqui MR, Tran K, Reddy SP, Malik AB. Reactive oxygen species in inflammation and tissue injury. Antioxid Redox Signal 2014; 20: 1126-1167
- 25 Viljoen A, Mncwangi N, Vermaak I. Anti-inflammatory iridoids of botanical origin. Curr Med Chem 2012; 19: 2104-2127
- 26 Horiuchi T, Mitoma H, Harashima S, Tsukamoto H, Shimoda T. Transmembrane TNF-alpha: structure, function and interaction with anti-TNF agents. Rheumatology 2010; 49: 1215-1228
- 27 Speranza L, Franceschelli S, Pesce M, Reale M, Menghini L, Vinciguerra I, De Lutiis MA, Felaco M, Grilli A. Antiinflammatory effects in THP-1 cells treated with verbascoside. Phytother Res 2010; 24: 1398-1404
- 28 Rutkowski L. Klucz do Oznaczania Roślin Naczyniowych Polski Niżowej. 2nd edition Warszawa: Wydawnictwo Naukowe PWN; 2011: 392
- 29 Tutin T, Heywood V, Burges N, Moore D, Valentine D, Walters S, Webb D. Flora Europaea. Vol. 3. Cambridge: Cambridge University Press; 1972: 148
- 30 Granica S, Bazylko A, Kiss AK. Determination of macrocyclic ellagitannin oenothein B in plant materials by HPLC-DAD-MS: method development and validation. Phytochem Anal 2012; 23: 582-587
- 31 Böyum A. A one-stage procedure for isolation of granulocytes and lymphocytes from human blood. General sedimentation properties of white blood cells in a 1 g gravity field. Scand J Clin Lab Invest Suppl 1968; 97: 51-76