Antihyperalgesic Effect of Eschscholzia Californica in Rat Models of Neuropathic Pain

Eschscholzia californica Cham. (Papaveraceae) is traditionally used by the Indians as a medicinal plant for its anxiolytic, anticonflict, analgesic and sedative properties. The mechanisms of action for the sedative and anxiolytic activities have not been clearly established and so to further investigate the pharmacological profile of E. californica in some painful conditions, a 70% v/v ethanol extract, DERnative=5:1, was tested in rat models of neuropathy induced by chronic constriction injury of the sciatic nerve (CCI), with chemotherapeutic oxaliplatin, and osteoarthritis caused by intrarticular injection of monoiodoacetate. In the CCI model evaluated in the rat paw-pressure test, the examined extract (100 mg kg−1 p.o.) showed an antihyperalgesic effect. Eschscholzia extract, after single injection at a dose of 100–300 mg kg−1 p.o., produced also a statistically significant decrease of pain perception on hyperalgesia induced by oxaliplatin and osteoarthritis, while in the same condition gabapentin did not display any antihyperalgesic effect. Furthermore, in the range of antihyperalgesic doses, the extract was efficacious in the hot-plate (thermal stimulus) and carrageenan tests (inflammatory model) without producing any behavioral impairment, as evaluated by the Irwin test. The analgesic effect exhibited by Eschscholzia extract in the mouse hot-plate test was not antagonized by naloxone, indicating that opioid neurotransmission is not involved in the effect. The above reported results suggest that a 70% (v/v) ethanol dried extract (DERnative=5:1) of E. californica might represent a promising product for the therapy of acute and chronic pain

An Integrated Approach to the Evaluation of a Metabolomic Fingerprint for a Phytocomplex. Focus on Artichoke [Cynara cardunculus subsp. scolymus] Leaf:

The availability of reliable herbal formulations is essential in order to assure the maximal activity and to limit unwanted side-effects. The correct concentration of declared components of herbal products is a matter of health legislation and regulation, but is still a topic under debate in the field of quality control assessment. In the present work specific constituents of artichoke leaf extracts, considered as a test herbal product, were measured by standard spectrophotometric and HPLC methods (for quantitative determination of some components only), and results were correlated with the ESI-MS (showing the full metabolomic fingerprint). Phytocomplex stability over time was also investigated in batches submitted to different storage conditions. The results indicated excellent agreement between the two approaches in the measurement of total caffeoylquinic acids and chlorogenic acid contents, but the metabolomic ESI-MS method approach provides a more complete evaluation and monitoring of the composition of a herbal product, without focusing only on a single/few compound measurements. Therefore, the ESI-MS method can be proposed for the evaluation of the quality of complex matrices, such as those in a phytocomplex. Another aspect lies in the possibility to obtain a broad-spectrum stability control of herbal formulations, requiring minimal sample pre-processing procedures

Efficacy of AdipoDren ® in Reducing Interleukin-1-Induced Lymphatic Endothelial Hyperpermeability

Lymphatic leakage can be seen as a detrimental phenomenon associated with fluid retention and deposition as well as gain of weight. Moreover, lymphatic dysfunction is associated with an inflammatory environment and can be a substrate for other health conditions. A number of treatments can ameliorate lymphatic vasculature: natural substances have been used as treatment options particularly suitable for their consolidated effectiveness and safety profile. Here we report the protective effect of AdipoDren®, an association of a series of plant-derived natural complexes, on lymphatic endothelium permeability promoted by interleukin-1 beta (IL-1β) and the associated molecular mechanisms. AdipoDren® demonstrated a protective effect on dermal lymphatic endothelial cell permeability increased by IL-1β. Reduced permeability was due to the maintenance of tight junctions and cell-cell localisation of occludin and zonula occludens-1 (ZO-1). Moreover, AdipoDren® reduced the expression of the inflammatory key element cyclooxygenase-2 (COX-2), while not altering the levels of endothelial and inducible nitric oxide synthases (eNOS and iNOS). The upregulation of antioxidant enzymatic systems (catalase and superoxide dismutase-1, SOD-1) and the downregulation of pro-oxidant markers (p22 phox subunit of NADPH oxidase) were also evident. In conclusion, AdipoDren® would be useful to ameliorate conditions of altered lymphatic vasculature and to support the physiological functionality of the lymphatic endothelium

The fingerprinting of Sedivitax, a commercial botanical dietary supplement: The classical LC-MS approach vs direct metabolite mapping.

The use of phytochemical preparations has shown a massive growth and consequently the related quality control is an important topic. Considering the possible interactions of the active molecules, arising to synergic phenomena, the use of the classical approach, based on the evaluation of the level of one ormore active compounds can be limitative.Recently a method, based on the direct infusion of commercial botanical dietary supplements in an ESI source operating in positive and negative ion modes has been proposed. The results so obtained were highly promising, allowing the presence of specific plant extracts in commercial products to be determined. In order to evaluate its validity, the data so obtained have been compared with those achievable by a more consolidated technique, as LCMS is. For this aim Sedivitax gocce (a commercial product composed by extracts of Passiflora incarnata, Eschscholtzia californica, Melissa officinalis and Valeriana officinalis) has been considered. Either the plant extracts or Sedivitax samples produced in different years have been analyzed by ESI (\ub1) with direct infusion and LC-MS. The data obtained were elaborated with different statistical methods. The results suggest that mass spectrometry linked to statistical methods can be a quick method to assess the overall stability of a botanical dietary supplement, and can be proposed as a promising perspective in quality control

A metabolite fingerprinting for the characterization of commercial botanical dietary supplements

Phytopharmaceuticals, phytomedicines and botanical dietary supplements are products of wide interest considering the increase of their use. The development of fast and effective analytical methods able to give a fingerprinting of the product, on the basis of the plant extracts declared to be contained in it, is surely of high interest. In a previous investigation electrospray mass spectrometry was proved to be effective for the characterization of plant extracts. The direct infusion of the samples and the analyses in both positive and negative ion mode lead to a clear differentiation of the different samples. To verify if the same approach can be effective also for mixtures of plant extracts, five different commercial dietary supplements [Sedivitax gocce (1), Finocarbo Plus opercoli (2), Sollievo Bio tavolette (3), MiniMas opercoli (4) and Ruscoven gocce (5), all products from Aboca S.p.A., Sansepolcro, Italy] were analyzed by ESI. In order to evaluate possible changes in the metabolic profile with respect to different years of production, ten different batches of the commercial dietary supplements were considered. The mass spectral data were evaluated by multivariate analysis and the obtained results suggest that the method allows a satisfactory and rapid characterization of complex mixtures of commercial dietary supplements

CAN STEVIA LEAF EXTRACTS PROTECT BETA CELLS AGAINST GLUCO- OR LIPOTOXICITY?

In the present study, the effects of leaf extracts of two Stevia rebaudiana Bertoni biotypes (namely ST2 and ST3) on beta cells exposed to gluco- or lipotoxic conditions have been evaluated. Leaf extracts were obtained through innovative and eco-friendly extraction technologies, able to obtain enriched fractions of desired bioactive compounds. Leaf phytochemical composition was spectrophotometrically assessed, in terms of total phenols and flavonoids as well as for their anti-radical and anti-oxidant activity, evaluated by DPPH radical-scavenging assay and ferric reducing antioxidant power (FRAP), respectively. Furthermore, the content and profile of the main steviol glycosides was analysed by HPLC, using an HILIC column under isocratic conditions, and quantified through standard curves of pure standard mixtures. MTT assay was used in order to evaluate the protection effect of stevia leaf extracts on beta cells exposed to gluco- or lipotoxicity conditions. INS-1E cells were exposed for 4 days to 30 mM glucose or 24h to 0.5 mM palmitate with or without increasing concentrations of stevia leaf extracts.The phytochemical characterisation revealed that ST2 had higher total phenolic and flavonoid content and antioxidant activity, in comparison with ST3. Conversely, ST3 showed higher (p<0.05) concentration of stevioside than ST2. Chronic exposure to high glucose concentration reduced INS-1E cells viability (-40%), that was completely prevented by exposure to 50 or 200 μg/mL ST2 and partially, but significantly, in the presence of ST3 genotype. Similarly, 24h exposure to palmitate resulted in a decrease of INS-1E viability (-20%), that was counteracted by both ST2 and ST3. The study shows that Stevia rebaudiana extracts have beneficial effects on viability of beta cells exposed to gluco- or lipotoxic conditions, possibly depending on their phytochemical composition and related antioxidant capacity

Protective effects of Stevia rebaudiana extracts on beta cells in lipotoxic conditions

Aims Stevia rebaudiana Bertoni leaf extracts have gained increasing attention for their potential protection against type 2 diabetes. In this study, we have evaluated the possible beneficial effects of Stevia rebaudiana leaf extracts on beta-cells exposed to lipotoxicity and explored some of the possible mechanisms involved. Methods Extracts, deriving from six different chemotypes (ST1 to ST6), were characterized in terms of steviol glycosides, total phenols, flavonoids, and antioxidant activity. INS-1E beta cells and human pancreatic islets were incubated 24 h with 0.5 mM palmitate with or without varying concentrations of extracts. Beta-cell/islet cell features were analyzed by MTT assay, activated caspase 3/7 measurement, and/or nucleosome quantification. In addition, the proteome of INS-1E cells was assessed by bi-dimensional electrophoresis (2-DE). Results The extracts differed in terms of antioxidant activity and stevioside content. As expected, 24 h exposure to palmitate resulted in a significant decrease of INS-1E cell metabolic activity, which was counteracted by all the Stevia extracts at 200 μg/ml. However, varying stevioside only concentrations were not able to protect palmitate-exposed cells. ST3 extract was also tested with human islets, showing an anti-apoptotic effect. Proteome analysis showed several changes in INS-1E beta-cells exposed to ST3, mainly at the endoplasmic reticulum and mitochondrial levels. Conclusions Stevia rebaudiana leaf extracts have beneficial effects on beta cells exposed to lipotoxicity; this effect does not seem to be mediated by stevioside alone (suggesting a major role of the leaf phytocomplex as a whole) and might be due to actions on the endoplasmic reticulum and the mitochondrion