Calceolarioside A, a Phenylpropanoid Glycoside from Calceolaria spp., Displays Antinociceptive and Anti-Inflammatory Properties.

Phenylpropanoid glycosides are a class of natural substances of plant origin with interesting biological activities and pharmacological properties. This study reports the antinociceptive and anti-inflammatory effects of calceolarioside A, a phenylpropanoid glycoside previously isolated from various Calceolaria species. In models of acute nociception induced by thermal stimuli, such as the hot plate and tail flick test, calceolarioside administered at doses of 1, 5, and 10 µg in the left cerebral ventricles did not modify the behavioral response of mice. In an inflammatory based persistent pain model as the formalin test, calceolarioside A at the high dose tested (100 µg/paw) reduced the licking activity induced by formalin by 35% in the first phase and by 75% in the second phase of the test. In carrageenan-induced thermal hyperalgesia, calceolarioside A (50 and 100 µg/paw) was able to significantly reverse thermal hyperalgesia induced by carrageenan. The anti-inflammatory activity of calceolarioside A was then assessed using the zymosan-induced paw edema model. Calceolarioside A (50 and 100 µg/paw) induced a significant reduction in the edema from 1 to 4 h after zymosan administration. Measuring IL-6, TNFα, and IL-1ß pro-inflammatory cytokines released from LPS-stimulated THP-1 cells, calceolarioside A in a concentration-dependent manner reduced the release of these cytokines from THP-1 cells. Taken together, our results highlight, for the first time, the potential and selective anti-inflammatory properties of this natural-derived compound, prompting its rationale use for further investigations.

Estrogens and phytoestrogens in body functions.

Estrogens are the hormones of reproduction in women as well as of many other important functions in the male and female body. They undergo significant changes in the different phases of life, e.g. during puberty, pregnancy or at menopause/andropause. Phytoestrogens are natural non-steroidal phenolic plant compounds that can mimic the activity of estrogens and their beneficial effects in women and in men. This narrative review summarizes the literature on the physiological role of estrogens and the several potential health benefits of phytoestrogens, with particular attention given to the possible role of phytoestrogens in aging.

Evaluation of in vitro cell and blood compatibility and in vivo analgesic activity of plant-derived dietary supplements.

OBJECTIVE: In vitro cell and blood compatibility of three dietary supplements, comprised of multiple plant extracts, Pneumo Go (PG), Green active (GA) and Equistasi (Eq), and their main component, the phytocomplex Matrix U.B.® (Union Bio S.r.l.) (M), were evaluated. Moreover, preliminary in vivo tests were performed on GA in order to assess its ability to reduce pain in an animal model. METHODS: Cell compatibility was determined using fibroblasts (NIH3T3) and primary adult human microvascular endothelial cells (HMVECad) and the neutral red uptake test. Blood compatibility was evaluated by analyzing blood parameters after incubation of the products with sodium citrate anticoagulated whole blood. Thrombin time was determined by adding thrombin to aliquots of human plasma containing the samples. Clotting time was revealed by an automatic coagulometer. The in vivo analgesic effect of GA was evaluated in Wistar rats using the formalin test. RESULTS: M and PG reduced the percentage of viable NIH3T3 cells, indicating their interference in the cell cycle. GA and Eq stimulated fibroblast proliferation and neutralized the toxic effect of M. M and PG reduced HMVECad cell viability. GA and Eq did not affect cell viability as well as negative control. The hemocompatibility tests indicated that all the samples did not interfere with fibrinogen. The in vivo test carried out in male rats showed a significant analgesic effect of GA in all formalin-induced pain behaviors. CONCLUSION: No hemotoxicity and good cell compatibility were found for all the tested samples. GA and Eq were the best candidates for further biocompatibility testing. Moreover, GA reduced pain in the animal model.

In vitro and in vivo characterization of the new analgesic combination Beta-caryophyllene and docosahexaenoic Acid.

Beta-caryophyllene (BCP) and docosahexaenoic acid (DHA) are components of several plants with documented anti-inflammatory and analgesic effects in animal pain models. In the present study, in vitro and in vivo tests were carried out to evaluate their effects, alone or in combination, during long-lasting administration in a model of persistent pain. IR spectra of the two compounds were obtained to determine their chemical stability and then in vitro toxicity was evaluated in fibroblasts and astrocytes. In the in vivo tests, the analgesic effects of BCP and BCP+DHA were determined in male rats subjected to a model of persistent recurrent pain (three repetitions of the formalin test once a week) to mimic recurrent pain. Both substances were administered per os in almond oil for 2 weeks. Gonadal hormones were determined at the end of the tests to evaluate treatment-induced effects on their levels. BCP changed fibroblast and astrocyte survival in a dose-dependent manner and the effect was counteracted by DHA coadministration. In the in vivo tests, pain responses were significantly decreased in the BCP and BCP+DHA groups with respect to OIL after 1 and 2 weeks of treatment. Estradiol and testosterone levels were increased only in the BCP group. In conclusion, BCP alone or at lower concentration in combination with DHA was efficacious in modulating pain, showing a clear analgesic activity.

Estrogenic chemicals at puberty change ERalpha in the hypothalamus of male and female rats.

The effects of two environmental endocrine disruptors, the synthetic pharmaceutical estrogen 17-ethinylestradiol (EE) and bisphenol-A (BPA), were analysed in male and female rats in a very sensitive developmental period, puberty. Immunohistochemistry was used to evaluate changes in the number of cells expressing estrogen receptors (ER-alpha) in the arcuate nucleus (ARC), ventromedial nucleus (VMH) and medial preoptic area (MPA) of the hypothalamus. Animals were treated during early puberty, from PND 23 to PND 30, with EE and BPA given orally every day. They were then sacrificed and perfused on PND 37 or PND 90, and blood and brains were collected for hormonal determination (testosterone and estradiol) and immunohistochemistry (estrogen receptors, ER). At PND 37, ER-labelled neurons were higher in males than in females in the ARC and MPA. EE and BPA increased ER-labelled neurons in the ARC and MPA. At PND 90, females showed higher ER-labelled neurons in the VMH. EE and BPA increased ER-labelled neurons in the MPA in females. EE increased testosterone in males at PND 37 and estradiol in females at PND 90. These results indicate the ability of estrogenic chemicals to change the reproductive neural circuits during puberty in male and female rats.

Effects of long-term exposure of lemon essential oil odor on behavioral, hormonal and neuronal parameters in male and female rats.

Behavioral, hormonal and neuronal responses to prolonged exposure to the volatile components of essential oil (EO) extracted from citrus lemon were investigated in male and female rats. Animals were exposed to the lemon essence for 2 weeks while in their cage. Anxiety was then determined with the elevated plus-maze apparatus while nociception was evaluated with a phasic thermal pain stimulus (plantar test) and with a chemical pain stimulus (formalin test). At the end of the experimental sessions, brain areas were dissected to measure beta-endorphin (beta-EP) concentrations in the hypothalamus and periaqueductal gray matter (PAG). Blood samples were collected to determine corticosterone plasma levels. In both sexes, prolonged EO exposure decreased the time spent in the open arms of the plus-maze apparatus. EO-exposed males and females showed higher thermal nociceptive thresholds than controls when tested with the plantar test apparatus. EO exposure induced female-specific decreases in formalin-induced pain behaviors during the formalin test. beta-EP concentrations in the hypothalamus and PAG were affected by EO. Corticosterone was lower in EO-exposed animals of both sexes than in their controls. These results suggest that long-term exposure to lemon EO can induce significant, at times sex-specific, changes in neuronal circuits involved in anxiety and pain.

Sex differences in the citrus lemon essential oil-induced increase of hippocampal acetylcholine release in rats exposed to a persistent painful stimulation.

The microdialysis technique was used to study the ability of essential oil from citrus lemon to modulate hippocampal acetylcholine (ACh) release in male and female rats. Animals were allowed to inhale this odor while experiencing a persistent nociceptive input (50 microl formalin, 5%) or under control conditions (sham-injection). In males, exposure to the essential oil did not change the time course and magnitude of the ACh increase induced by pain. In females, the pain-induced increase of ACh was delayed and increased by exposure to lemon essential oil. The present results indicate that lemon essential oil affects the ACh release differently in male and female rats during a painful condition.