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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.

Lamium album and Lamium purpureum are species belonging to the genus Lamium. Aerial parts of the two species and roots of Lamium album have applications in human and veterinary traditional medicine. Haemostatic properties of butanolic extracts of Lamium species were investigated by two experimental models in Wistar rats: haemostatic test by tail bleeding time determination and acenocoumarolcarrageenan test. Results of the haemostatic test by tail bleeding determination demonstrated haemostatic activity of both extracts. In the acenocoumarol-carrageenan test, only the Lamium album extract showed haemostatic activity, comparable to that of vitamin K. Based on the qualitative chemical data on iridoid glycosides (HPTLC), 8-acetylshanzhiside methyl ester might be assumed to be responsible for haemostatic activity. Based on the acute toxicity test, none of the extracts showed toxicity.

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.

This work was aimed at testing gastroprotective effects of Centaurea cyanus L. (herba) polysaccharides (P) and polyphenols (A) fractions on stress-induced rat ulcer model. Studies evaluating acute toxicity in rats and antioxidant (chemiluminescence method) and antimicrobial (on Staphylococcus aureus ATCC6538 and Escherichia coli ATCC8739 strains) activities of Centaurea cyanus L. (herba) product, which combined polysaccharides and polyphenols fractions (PA), have also been done. Accordingly, in vivo pharmacological studies revealed high influence of PA product (500 mg kg-1 of body weight) on deep, moderate and superficial gastric mucosal lesions, greater than that of chemical reference, Ranitidine. P product was proven more effective than Ranitidine in opposing the emergence of deep necrotic lesions only, suggesting the ability of polysaccharides compounds to consolidate gastric mucous layer as well as their certain tendency for cooperation with polyphenols fractions. The acute toxicity study indicated the lack of toxicity of PA product and maximum tolerated doses greater than 1875 mg kg-1 of body weight. PA product provided augmented scavenger activity and week antimicrobial activity on Staphylococcus aureus ATCC6538 and Escherichia coli ATCC8739 strains, resulting in better opportunities for valorisation of the aerial part of Centaurea cyanus L. species in order to obtain new and effective natural medicines.

High-mobility group box protein 1 (HMGB1) is an intracellular protein that may be released actively from monocytes and macrophages or passively from necrotic or damaged cells. Its inhibition in animal experiments, even in the late phase of septic shock, significantly enhanced the survival rate of rodents. The aim of our study was to investigate the effect of a vegetal fraction isolated and highly purified from Helleborus purpurascens regarding the modulation of HMGB1 release either from tumor cells or human blood mononuclear cells. Our results showed that the vegetal fraction was able to down-regulate the release of HMGB1 from activated human blood mononuclear cells (PBMCs) and tumor cells. By combining the purified fraction with Cyclophosphamide the release of HMGB1 from tumor cells was strongly decreased. This synergism was not noticed when the ve getal product was associated with Doxorubicin. We also studied the effect of the purified fraction in mice with septic shock induced by cecal ligation and puncture (CLP) method. The tested vegetal product increased significantly the survival rate of animals compared to the mice not treated with it. Our data suggest that the purified vegetal fraction may modulate inflammation by down-regulating the HMGB1, which can also explain its efficacy in septic shock in mice.