Studi preclinici su piante medicinali utilizzate nel trattamento delle patologie prostatiche

La fitoterapia viene spesso impiegata nel trattamento e prevenzione di patologie prostatiche di grado lieve-moderato. L’eziopatogenesi delle prostatiti, dell’iperplasia e dell’ipertrofia prostatica benigna (IPB) può essere complessa e l’intervento è spesso multi-target. Scopo del presente elaborato è stato quello di analizzare i meccanismi d’azione delle piante medicinali più comunemente utilizzate. Le proprietà farmacologiche, esplicate a livello delle vie urinarie, più ricorrenti sono: anti-androgenica, antiestrogenica, anti-proliferativa, antiossidante ed antinfiammatori

Evaluation of in vitro neurotoxicity of methyl mercury, PCB153 and PCB126

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Antimutagenic and antioxidant activity of a protein fraction from aerial parts of Urtica dioica

Abstract Context: Urtica dioica L. (Urticaceae), stinging nettle, has been employed as a folklore remedy for a wide spectrum of ailments, including urinary disorders, prostatic hyperplasia, and liver diseases. It has been also used traditionally for cancer treatment. Object: To evaluate the potential chemopreventive properties of a protein fraction from the aerial part of Urtica dioica (namely UDHL30). Materials and methods: UDHL30 has been tested for the antimutagenic activity in bacteria (50-800 μg/plate; Ames test by the preincubation method) and for the cytotoxicity on human hepatoma HepG2 cells (0.06-2 mg/mL; 24 and 48 h incubation). Moreover, the antioxidant activity of UDHL30 (0.1-1200 μg/mL; ABTS and superoxide-radical scavenger assays) was evaluated as potential protective mechanisms. Results: UDHL30 was not cytotoxic on HepG2 cells up to 2 mg/mL; conversely, it exhibited a strong antimutagenic activity against the mutagen 2-aminoanthracene (2AA) in all strains tested (maximum inhibition of 56, 78, and 61% in TA98, TA100, and WP2uvrA strains, respectively, at 800 μg/plate). In addition, a remarkable scavenging activity against ABTS radical and superoxide anion (IC50 values of 19.9 ± 1.0 μg/mL and 75.3 ± 0.9 μg/mL, respectively) was produced. Discussion and conclusions: UDHL30 possesses antimutagenic and radical scavenging properties. Being 2AA a pro-carcinogenic agent, we hypothesize that the antimutagenicity of UDHL30 can be due to the inhibition of CYP450-isoenzymes, involved in the mutagen bioactivation. The radical scavenger ability could contribute to 2AA-antimutagenicity. These data encourage further studies in order to better define the potential usefulness of UDHL30 in chemoprevention

β-caryophyllene and low-doses of doxorubicin against liver cancer cells: a “metronomic chemotherapy”

Cholangiocarcinoma and hepatocellular carcinoma are primary liver cancers, both representing a growing challenge due to their increasing morbidity and mortality. A “metronomic chemotherapy”, consisting of the repeated administration of low and/or continuous doses of anti-neoplastic drugs, represents an alternative approach to the standard chemotherapy [1]. Numerous natural substances exhibited in vitro chemosensitizing features: in particular, the natural sesquiterpene β-caryophyllene (CRY) has been proved to increase the cytotoxicity of doxorubicin (DOXO) in leukemic cells [2]. Hence, our aim has been to evaluate the ability of CRY to enhance the efficacy of low-dose DOXO in human liver cancer cells, by applying a metronomic protocol. To this end, human liver HepG2 and CCA cells have been used as models of hepatocellular carcinoma and cholangiocarcinoma. The metronomic protocol was based on a 2h low-time exposition to the test substances, followed by 72h incubation for restoring. This scheduling has been applied 3 times and cytotoxicity was measured by MTT assay. Both the substances alone (CRY 1-100 μg/ml; DOXO 1-500 μg/ml) and the combination of DOXO with a nontoxic concentration of CRY were assessed. We found that the repeated treatments with low concentrations produced a significant potentiation (about 30 %) of DOXO cytotoxicity in HepG2. The combination with CRY increased the DOXO activity, reaching a 70 % inhibition of cell viability at 50 μg/ml after 2 repeated treatments. Similar effects were found in CCA, although repeated treatments induced no additional potentiation. These results highlight a possible role of CRY as a chemosensitizing agent for DOXO-based chemotherapy of liver cancer

β-caryophyllene and low-doses of doxorubicin against liver cancer cells: a “metronomic chemotherapy”

Cholangiocarcinoma and hepatocellular carcinoma are primary liver cancers, both representing a growing challenge due to their increasing morbidity and mortality. A “metronomic chemotherapy”, consisting of the repeated administration of low and/or continuous doses of anti-neoplastic drugs, represents an alternative approach to the standard chemotherapy [1]. Numerous natural substances exhibited in vitro chemosensitizing features: in particular, the natural sesquiterpene β-caryophyllene (CRY) has been proved to increase the cytotoxicity of doxorubicin (DOXO) in leukemic cells [2]. Hence, our aim has been to evaluate the ability of CRY to enhance the efficacy of low-dose DOXO in human liver cancer cells, by applying a metronomic protocol. To this end, human liver HepG2 and CCA cells have been used as models of hepatocellular carcinoma and cholangiocarcinoma. The metronomic protocol was based on a 2h low-time exposition to the test substances, followed by 72h incubation for restoring. This scheduling has been applied 3 times and cytotoxicity was measured by MTT assay. Both the substances alone (CRY 1-100 μg/ml; DOXO 1-500 μg/ml) and the combination of DOXO with a nontoxic concentration of CRY were assessed. We found that the repeated treatments with low concentrations produced a significant potentiation (about 30 %) of DOXO cytotoxicity in HepG2. The combination with CRY increased the DOXO activity, reaching a 70 % inhibition of cell viability at 50 μg/ml after 2 repeated treatments. Similar effects were found in CCA, although repeated treatments induced no additional potentiation. These results highlight a possible role of CRY as a chemosensitizing agent for DOXO-based chemotherapy of liver cancer

Genotoxicity assessment of piperitenone oxide: an in vitro and in silico evaluation

Piperitenone oxide, a natural flavouring agent also known as rotundifolone, has been studied for the genotoxicity assessment by an integrated in vitro and in silico experimental approach, including the bacterial reverse mutation assay, the micronucleus test, the comet assay and the computational prediction by Toxtree and VEGA tools. Under our experimental conditions, the monoterpene showed to induce both point mutations (i.e. frameshift, base-substitution and/or oxidative damage) and DNA damage (i.e. clastogenic or aneuploidic damage, or single-strand breaks). Computational prediction for piperitenone oxide agreed with the toxicological data, and highlighted the presence of the epoxide function and the α,β-unsaturated carbonyl as possible structural alerts for DNA damage. However, improving the toxicological libraries for natural occurring compounds is required in order to favour the applicability of in silico models to the toxicological predictions. Further in vivo evaluations are strictly needed in order to evaluate the role of the bioavailability of the substance and the metabolic fate on its genotoxicity profile. To the best of our knowledge, these data represent the first evaluation of the genotoxicity for this flavour compound and suggest the need of further studies to assess the safety of piperitenone oxide as either flavour or fragrance chemicals

Role of Cassia angustifolia on hepatic toxicity in Wistar rats

Background. Cassia angustifolia L. (senna) is traditionally used as a laxative for the short-term treatment of acute constipation and as a purgative before diagnostic endoscopy. Among several adverse reactions in literature, hepatotoxicity (Sonmez et al., Gastroenterol. Belg., 2005;68:385-7) and portal vein thrombosis (Soyuncu et al., Clin. Toxicol. Phila, 2008;46:774-7) have been reported. Present study was aimed to evaluate the long-term effects of C. angustifolia by both in vivo (oral administration to rats) and in vitro (liver cell cultures) approaches. Materials and methods. In vivo, experiments were performed in Wistar rats, after oral administration of senna leaf extract (12 and 58 mg/kg/day) for 4 or 8 weeks. Serum was used for biochemical analysis. Liver samples were used for hystomorphological and immunohistochemical examination (Gaudio E. et al., Gastroenterology. 2006;130:1270-82) along with the determination of oxidative stress parameters. Cytotoxicity was assessed on Buffalo normal Rat Liver cell line (BRL-3A) by the Trypan blue assay and the MTT reduction method after 24 h of exposure. Results. In Wistar rats the extract did not induce any significant change in animal body weight, food and water consumption, enzyme activities, hystomorphological hepatic characteristics, levels of reduced glutathione, and MDA formation, at either time or dosage level. In BRL-3A cells the substance induced concentration-dependent cytotoxicity. Conclusions. C. angustifolia, at doses about 10 and 50 times higher than those used in humans (during a lapse of time corresponding to a chronic administration) does not affect liver morphology and hepatic function indices in rats. In vitro senna extract is cytotoxic at concentrations at least 2000 fold higher than those obtainable in vivo. Nevertheless, in humans the safety of C. angustifolia should be further monitored, in terms of patient-related factors

Role of acute administration of Chelidonium majus alone and in association with acetaminophen in rat liver

Aerial parts of Chelidonium majus (CM) are used to treat liver and gallbladder disorders, however several cases of hepatotoxicity following use of CM preparations are reported (Moro et al, 2009). Previous studies showed that high doses of CM do not alter morpho-functional rat liver parameters (Mazzanti et al. 2009). We aimed to evaluate if the co-treatment of CM and sub-toxic doses of acetaminophen can induce rat liver damage. Rats were treated as follows: 1) received vehicle at 0, 12, 24, 36 and 37h; 2) received vehicle at 0, 12, 24, 36 vehicle and acetaminophen at 37h; 3) received CM at 0, 12, 24, 36 and vehicle at 37h; 4) received CM at 0, 12, 24, 36 and acetaminophen at 37h. 24 hours after last treatment animals were sacrified and blood samples were collected to perform biochemical analysis and liver samples were underwent histomorphological and immunohistochemical examination. CM or acetaminophen did not alter body weight whereas total bilirubin, AST and ALT were slightly modified. Neither CM nor acetaminophen altered the hepatic structure while both substances induced a weak inflammatory cells infiltration. The co-administration of CM and acetaminophen did not modify the picture caused by the single treatments. Expression of α-SMA in hepatic stellate cells was negative in all experimental groups. In conclusion, the co-administration of C.majus and acetaminophen, does not have any synergistic effect in this experimental model. In humans, the safety of CM should be further monitored, taking into account the possibility of idiosyncratic reactions and/or other patient-related factors

A polyphenol rich extract from Solanum melongena L. DR2 peel exhibits antioxidant properties and anti-herpes simplex virus type 1 activity in vitro

DR2B and DR2C extracts, obtained by ethanolic maceration of peel from commercially and physiologically ripe aubergine berries, were studied for the antioxidative cytoprotective properties and anti-HSV-1 activity, in line with the evidence that several antioxidants can impair viral replication by maintaining reducing conditions in host cells. The antioxidative cytoprotective effects against tBOOH-induced damage were assessed in Caco2 cells, while antiviral activity was studied in Vero cells; polyphenolic fingerprints were characterized by integrated phytochemical methods. Results highlighted different compositions of the extracts, with chlorogenic acid and delphinidin-3-rutinoside as the major constituents; other peculiar phytochemicals were also identified. Both samples reduced reactive oxygen species (ROS) production and exhibited scavenging and chelating properties. DR2C partly counteracted the tBOOH-induced cytotoxicity, with a remarkable lowering of lactate metabolism under both normoxia and hypoxia; interestingly, it increased intracellular GSH levels. Furthermore, DR2C inhibited the HSV-1 replication when added for 24 h after viral adsorption, as also confirmed by the reduction of many viral proteins’ expression. Since DR2C was able to reduce NOX4 expression during HSV-1 infection, its antiviral activity may be correlated to its antioxidant properties. Although further studies are needed to better characterize DR2C activity, the results suggest this extract as a promising new anti-HSV-1 agent

Analysis of Safety Concerns on Herbal Products with Assumed Phytoestrogenic Activity

Phytoestrogens (PEs) are plant-based compounds that can interact with estrogen receptors and are mainly used to treat menopausal complaints. However, the safety of products with assumed phytoestrogenic activity is not fully understood. This study aimed to identify plant species with assumed phytoestrogenic activity, review existing literature on their use and safety, and critically evaluate adverse reaction (AR) reports of single-herb, multi-herb, and mixed-multiple products, as submitted to the Netherlands Pharmacovigilance Centre Lareb and to VigiBase of the World Health Organization (WHO). In the Lareb database, the most commonly reported plant species to cause ARs (total of 67 reports) were Actaea racemosa L. (black cohosh) (47.8%), Humulus lupulus L. (hops) (32.8%), and Glycine max (L.) Merr. (soybean) (22.4%). In the VigiBase database (total of 21,944 reports), the top three consisted of Glycine max (L.) Merr. (71.4%), Actaea racemosa L. (11.6%), and Vitex agnus-castus L. (chaste tree) (6.4%). In the scoping review (total of 73 articles), Actaea racemosa L. (30.1%), Glycine max (L.) Merr. (28.8%), and Trifolium pratense L. (13.7%) were the most frequently mentioned plant species. ARs were most frequently reported in the system organ classes "gastrointestinal disorders", "skin and subcutaneous tissue disorders", "reproductive system and breast disorders", and "general disorders and administration site conditions". Furthermore, from the scoping review, it appeared that the use of products with assumed phytoestrogenic activity was associated with postmenopausal bleeding. It was concluded that, while the potential benefits of products with assumed phytoestrogenic activity have been extensively pursued, the potential occurrence of ARs after using these products is less well understood. This study highlights the need for further investigation and careful monitoring of these products to better understand their effects and ensure the safety and well-being of individuals using them. </p