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(1) Background: Helleborus purpurascens Waldst. &amp, Kit. (hellebore) is a plant species found mainly in Balkans and the Carpathians, and it is traditionally used for a variety of ailments since the time of Hippocrates. The aim of this study was to investigate the immunomodulatory effect of hellebore extracts correlated with relevant chemical compounds and the extraction method. (2) Methods: A methanolic (H1) and a hydroalcoholic extract (H2) were prepared by standard methods. Qualitative (HPTLC) and quantitative (HPLC) chemical analysis were conducted to reveal the ecdysones and polyphenolic compounds. In vitro studies were performed using rat macrophages, murine fibroblasts and immortalized human T-lymphocytes, and their viability was determined by MTS assay. In vivo studies involved a rat immunodepression model. (3) Results: In vitro assays revealed the stronger effect of H2 on cellular proliferation, compared to H1. In the in vivo assay, H2 revealed an immunostimulatory effect in the context of experimentally induced immunosuppression with dexamethasone, a superior effect to levamisole treatment according to the same regimen, in two doses every 24 h. There was no correlation between pharmacological effect and the reference compounds evaluated. (4) Conclusions: The immunomodulatory effect of methanolic and hydroalcoholic hellebore extracts is not due to ecdysones and polyphenolic compounds, but other polar substances, possible steroid glycosides.

Linking Europe and Central Asia, Romania is considered to be an important meeting point between different biogeographic regions and ecosystems. In this context, Romanian flora is truly an exceptional source of biological active phytocompounds, basis of new biopharmaceuticals. Data indicate that in Romania there are about 3600 higher plants (ferns and flowering plants) that grow wild and other 800 hybrid species. Amongst these, about 800 are medicinal species, some of them unique in Europe. In order to demonstrate their potential use as active ingredients in biopharmaceutical engineering, this work was aimed at revealing chemical composition, particularly polyphenols profile (HPTLC method) and antioxidant activity (DPPH method) face to potential dermal benefits (single and repeated dose tolerance test in rabbits) of two (ethanolic) extracts prepared from some less studied plant pieces of two valuable phytomedicines; leaves of chokeberries (Aronia melanocarpa L.) and leaves of sea-buckthorn (Hippophae rhamnoides L.). Analytical studies have shown the occurrence of numerous polyphenols species, quercetin, gallic and caffeoylquinic acid derivates, known to have a high antioxidant potency. DPPH assay has confirmed high antioxidant activity of both vegetal extracts, even stronger than that of references, rutin and gallic acid phenolics (IC50 ranging from 2.66 to 5.08 μg / mL). In vivo, pharmacological studies have indicated good dermal tolerance and healthy, supple and reepitelized tegument after being treated with both vegetal extracts, which has revealed their potential use as antioxidant and healing ingredients for (topic) biopharmaceuticals. Thus, while Aronia melanocarpa L. as well as Hippophae rhamnoides L. are known for their valuable fruit and seed derived products, this study proves that the (ethanolic) extracts prepared from their leaves pieces should also be taken into consideration.

Verbascum phlomoides L. (Scrophulariaceae) (mullein) used in the European folk medicine due to its anti-inflammatory and soothing action on the respiratory tract is thoroughly documented in handbooks and scientific literature. Nevertheless, information regarding the influence of the polyphenol content on pharmacological activity is scarce.This study explored the antioxidant and anti-inflammatory potential of V. phlomoides polyphenol-rich extract.Dried mullein flowers (200 g) were subjected to water extraction (60 °C, 2 h, herb/solvent ratio = 1/10 m/v) and further to n-butanol partition. Total phenolics were spectrophotometrically determined and specific compounds were evaluated by HPLC. The antioxidant activity was assessed by the 2,2-di(4-tert-octylphenyl)-1-picrylhydrazyl (DPPH) assay. The anti-inflammatory potential of the extract (50-200 μg/mL) was evaluated in vitro by ELISA measurement of ICAM-1 expression in TNF-α-stimulated endothelial cells and in vivo by the rat paw edema assay.The mullein extract contained 4.18% total polyphenols expressed as gallic acid. The main components identified by HPLC were: rosmarinic acid (14.93 mg/g), caffeic acid (39.96 mg/g), ferulic acid (29.61 mg/g) and quercetin (17.29 mg/g). Acteoside was not detected; aucubin was detected in traces (0.028 mg/g). Depending on concentration, the extract exerted scavenging activity on DPPH radical (EC50 7.09 mg/mL), significantly inhibited TNF-α-induced ICAM-1 expression by 55-58.8% on human umbilical vein endothelial cells at 100 and 200 μg/mL, but failed to reduce egg-white-induced rat paw edema.Mullein polyphenols play an important role in exerting the antioxidant effect but have a weak influence on anti-inflammatory activity that is correlated, probably, to a higher content of iridoids and phenylethanoids.