Anti-inflammatory, Gastroprotective, and Cytotoxic Effects ofSideritis scardicaExtracts

Abstract

Sideritis scardica Griseb. (ironwort, mountain tea), an endemic plant of the Balkan Peninsula, has been used in traditional medicine in the treatment of gastrointestinal complaints, inflammation, and rheumatic disorders. This study aimed to evaluate its gastroprotective and anti-inflammatory activities. Besides, continuously increasing interest in assessing the role of the plant active constituents preventing the risk of cancer was a reason to make a detailed examination of the investigated ethanol, diethyl ether, ethyl acetate, and N-butanol extracts regarding cytotoxicity. Oral administration of the investigated extracts caused a dose-dependent anti-inflammatory effect in a model of carrageenan-induced rat paw edema. Gastroprotective activity of the extracts was investigated using an ethanol-induced acute stress ulcer in rats. The cytotoxic activity of plant extracts was assessed on PBMC, B16, and HL-60 cells and compared to the cytotoxicity of phenolic compounds identified in extracts. Apoptotic and necrotic cell death were analyzed by double staining with fluoresceinisothiocyanate (FITC)-conjugated annexin V and PI. The developed HPLC method enabled qualitative fingerprint analysis of phenolic compounds in the investigated extracts. Compared to the effect of the positive control, the anti-inflammatory drug indomethacine (4 mg/kg), which produced a 50 % decrease in inflammation, diethyl ether and N-butanol extracts exhibited about the same effect in doses of 200 and 100 mg/kg (53.6 and 48.7 %; 48.4 and 49.9 %, respectively). All investigated extracts produced dose-dependent gastroprotective activity with the efficacy comparable to that of the reference drug ranitidine. The diethyl ether extract showed significant dose-dependent cytotoxicity on B16 cells and HL-60 cells, decreasing cell growth to 51.3 % and 77.5 % of control, respectively, when used at 100 µg/mL. It seems that phenolic compounds (apigenin, luteolin, and their corresponding glycosides) are responsible for the diethyl ether extract cytotoxic effect. It also appears that induction of oxidative stress might be involved in its cytotoxicity, since B16 and HL-60 cells increased their ROS production in response to treatment with diethyl ether extract. Neither of the tested extracts nor any phenolic compounds showed significant cytotoxic effect to human PBMC. These results demonstrated the potent anti-inflammatory and gastroprotective activities, as well as the promising cytotoxicity.

Assessment of the Acute and Subchronic Toxicity and Mutagenicity of Sideritis scardica Griseb. Extracts

“…Until now, there has been no comprehensive data on the toxicity of S. scardica except for one in vitro study, in which a significant dose-dependent decrease in proliferation was found in B16 (by 51.3%) and HL-60 (by 77.5%) cells treated with diethyl ether extract, and as speculated, phenolic compounds such as apigenin, luteolin, and their corresponding glycosides could be responsible for this effect [18]. These findings strongly advocated the need to further the toxicity evaluation of S. scardica , preferably using an in vivo experimental model.…”

“…The extracts were analyzed for total polyphenols with a Folin–Ciocalteu method following guidelines of the European Pharmacopoeia [18] and for specific polyphenolic compounds (flavonoids, acteoside, caffeoylquinic acids) with a high-performance liquid chromatography (HPLC) method. For this purpose, a LunaR C18/2 column (Phenomenex, Torrance, CA, USA; 250 mm length, 4.6 mm inner diameter, 5 μm particle size) was used at 40 °C in a Shimadzu LC10 HPLC system (Shimadzu Deutschland GmbH, Duisburg, Germany).…”

“…This is particularly relevant for data on the acute and repeated-dose toxicity and mutagenicity. The only available data is from the in vitro studies involving B16 cells and HL-60 cells, and indicates that highly lipophilic diethyl ether extract of S. scardica can exert moderate cytotoxic effects [18]. Considering that wild specimens of S. scardica are collected for consumption and that there is a growing evidence of the potential therapeutic use of its extracts in some neurodegenerative disorders, it is imperative to evaluate whether constituents of this plant do not exhibit health threats.…”

Morphological, chemical and pharmacological characterization of selected plant species of genus Arbutus L., Bruckenthalia Rchb., Calluna Salisb. i Erica L. (Ericaceae)

“…Nakon 3h se razvija intenzivna fagocitna inflamacija sa velikim brojem neutrofila i otokom tkiva. Inhibicija edema predstavlja dokaz antiinflamatornog dejstva (158,159).…”

“…Brojna istraživanja su pokazala da mnoge aktivne komponente biljaka, pre svega flavonoidi i fenolkarbonske kiseline, sprečavaju infiltraciju neutrofila u inflamirano područje i uklanjaju slobodne radikale pa deluju antiinflamatorno. Flavonoidi mogu da inhibiraju 5-lipooksigenazu i ciklooksigenazu, a deluju i kao inhibitori PLA2, pa utiču na biosintetske puteve prostanoida (63,156,159).…”