12 research outputs found
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
Treatment with tocilizumab or corticosteroids for COVID-19 patients with hyperinflammatory state: a multicentre cohort study (SAM-COVID-19)
Objectives: The objective of this study was to estimate the association between tocilizumab or corticosteroids and the risk of intubation or death in patients with coronavirus disease 19 (COVID-19) with a hyperinflammatory state according to clinical and laboratory parameters.
Methods: A cohort study was performed in 60 Spanish hospitals including 778 patients with COVID-19 and clinical and laboratory data indicative of a hyperinflammatory state. Treatment was mainly with tocilizumab, an intermediate-high dose of corticosteroids (IHDC), a pulse dose of corticosteroids (PDC), combination therapy, or no treatment. Primary outcome was intubation or death; follow-up was 21 days. Propensity score-adjusted estimations using Cox regression (logistic regression if needed) were calculated. Propensity scores were used as confounders, matching variables and for the inverse probability of treatment weights (IPTWs).
Results: In all, 88, 117, 78 and 151 patients treated with tocilizumab, IHDC, PDC, and combination therapy, respectively, were compared with 344 untreated patients. The primary endpoint occurred in 10 (11.4%), 27 (23.1%), 12 (15.4%), 40 (25.6%) and 69 (21.1%), respectively. The IPTW-based hazard ratios (odds ratio for combination therapy) for the primary endpoint were 0.32 (95%CI 0.22-0.47; p < 0.001) for tocilizumab, 0.82 (0.71-1.30; p 0.82) for IHDC, 0.61 (0.43-0.86; p 0.006) for PDC, and 1.17 (0.86-1.58; p 0.30) for combination therapy. Other applications of the propensity score provided similar results, but were not significant for PDC. Tocilizumab was also associated with lower hazard of death alone in IPTW analysis (0.07; 0.02-0.17; p < 0.001).
Conclusions: Tocilizumab might be useful in COVID-19 patients with a hyperinflammatory state and should be prioritized for randomized trials in this situatio
Melanzana rossa di Rotonda: analisi fitochimica e attività antiossidante
Obiettivo del presente progetto di tesi è stato valutare le caratteristiche potenzialmente funzionali della “Melanzana Rossa di Rotonda”, un ecotipo lucano di Solanum aethiopicum L. che ha ottenuto il riconoscimento europeo della “denominazione di origine protetta” (DOP), a sottolineare lo stretto nesso tra le particolari caratteristiche del prodotto ed il territorio in cui si coltiva. Sia il frutto che le foglie di questa specie hanno un uso consolidato in gastronomia. Nel presente studio, è stata valutata la potenziale attività antiossidante della melanzana rossa di Rotonda DOP, in relazione alla sua composizione fitochimica, con particolare riferimento al contenuto in polifenoli. Da campioni freschi di frutto e di foglie, certificati dal consorzio di produzione ALSIA, sono stati ottenuti gli estratti etanolici di buccia (MB), polpa (MP), frutto in toto (MFr) e foglie (MFo). Sugli estratti è stata condotta l’analisi del fingerprinting metabolomico, mediante cromatografia HPTLC e densitometria, la determinazione dei polifenoli totali (test di Folin-Ciocalteau) e di flavonoidi (test di Down) e test di attività antiossidante (Gulcin, 2012), sia diretta che indiretta (attività scavenger dei radicali DPPH, ABTS, ossidrile e anione superossido; inibizione della perossidazione lipidica; attività chelante e riducente degli ioni ferro). La composizione fitochimica della melanzana rossa DOP è stata confrontata anche con quella di due varietà lucane di melanzana comune, in particolare S. melongena var. bianca di Senise e S. melongena var. purposa.
L’analisi fitochimica ha evidenziato una peculiare composizione in polifenoli della melanzana rossa DOP, rispetto alle melanzane comuni di riferimento. Tali differenze erano particolarmente evidenti nel fingerprint di MB ma si riflettevano anche in quello di MFr. Tra i polifenoli, l’acido clorogenico era presente in maniera ubiquitaria nei vari estratti saggiati, mentre acido caffeico ed apigenina erano presenti soltanto negli estratti di melanzana rossa. Inoltre, la concentrazione dell’acido clorogenico era bassa nella polpa della melanzana rossa: questo potrebbe essere responsabile dello lento imbrunimento della stessa dopo il taglio. I dati quantitativi ottenuti dai test di Folin-Ciocalteau e Down evidenziavano la presenza ubiquitaria dei polifenoli nei vari estratti saggiati e la composizione peculiare in flavonoidi degli estratti MB ed MFo. Nei test di attività antiossidante, tutti gli estratti di melanzana rossa risultavano attivi come scavenger dei radicali DPPH, ABTS, e delle specie ROS (radicale idrossilico e anione superossido), come inibitori della perossidazione lipidica e come chelanti degli ioni ferrosi e ferrici, anche se con potenza differente. Gli estratti MB ed MFo erano in generale più potenti di MP ed MFr e, in alcuni casi, MFo produceva effetti antiossidanti confrontabili con quelli dei controlli positivi di riferimento.
I risultati ottenuti mostrano, pertanto, che la melanzana rossa di Rotonda DOP possiede una spiccata attività antiossidante da ascrivere probabilmente al contenuto in polifenoli, in particolare flavonoidi.
L’evidenza di dati scientifici circa il potenziale ruolo funzionale di questo prodotto, sebbene richieda validazioni in sistemi biologici, potrebbe favorirne lo sviluppo futuro come supplemento della dieta ed incrementarne la coltivazione e la produzione, con evidenti risvolti positivi sull’economia lucana.
Bibliografia
Gulcin I. (2012), “Antioxidant activity of food constituents: an overview”, Arch. Toxicol., 86: 345–391
Chemopreventive effects of β-caryophyllene against cigarette smoke damage in upper airway cells
Exposure to cigarette smoke induces damages in different organs and tissues, and high incidence of precancerous lesions and malignancies particularly at upper respiratory tract.1 Increasing levels of reactive oxygen species (ROS) and the activation of STAT3 pathway seems to be involved in smoke injury and oncogenic proliferation of damaged cells.1 In order to find new strategies for preventing cancer development in smokers, in present study we evaluated the ability of the natural sesquiterpene β-caryophyllene (CRY) to inhibit smoke damage in human epithelial bronchial upper airway (BEAS-2B) cells, by studying the inhibition of STAT3-phosphorylation, pro-oxidant damage and oncogenic proliferation induced by a condensed sample of cigarette smoke (CSC), according to previous methods.2,3 In our experiments, CSC (25-150 g/ml) strongly increased the levels of both phosphorylated STAT3 and intracellular ROS, and the cell migration capacity. CRY (1-10 g/ml) significantly reduced the activation of STAT3 pathway and the pro-oxidant effects of CSC (75 μg/ml). Furthermore, a remarkable inhibition of CSC-induced cell migration was highlighted, so suggesting a possible interference of CRY with metastatic ability of damaged cells. Data obtained highlight interesting protective properties of CRY and encourage further studies in order to evaluate its possible use as a chemopreventive agent against smoke damage.
References
1. Wu et al. (2014). Free Rad. Biol. Med. 69, 208–218.
2. Chichiarelli et al. (2010). Arch Biochem Biophys 494, 178-193.
3. Duan et al. (2013). Plos One 8, e57941
SPC Liposomes as Possible Delivery Systems for Improving Bioavailability of the Natural Sesquiterpene β-Caryophyllene: Lamellarity and Drug-Loading as Key Features for a Rational Drug Delivery Design
The natural sesquiterpene β-caryophyllene (CRY) has been highlighted to possess interesting pharmacological potentials, particularly due to its chemopreventive and analgesic properties. However, the poor solubility of this sesquiterpene in aqueous fluids can hinder its uptake into cells, resulting in inconstant responses of biological systems, thus limiting its application. Therefore, identifying a suitable pharmaceutical form for increasing CRY bioavailability represents an important requirement for exploiting its pharmacological potential. In the present study, the ability of soybean phosphatidylcholine (SPC) liposomes to improve bioavailability and absorption of CRY in cancer cells has been evaluated. Liposomal formulations of CRY, differing for lamellarity (i.e., unilamellar and multilamellar vesicles or ULV and MLV) and for the drug loading (i.e., 1:0.1, 1:0.3 and 1:0.5 mol/mol between SPC and CRY) were designed with the aim of maximizing CRY amount in the liposome bilayer, while avoiding its leakage during storage. The low-loaded formulations significantly potentiated the antiproliferative activity of CRY in both HepG2 and MDA-MB-468 cells, reaching a maximum IC50 lowering (from two to five folds) with 1:0.3 and 1:0.1 SPC/CRY MLV. Conversely, increasing liposome drug-loading reduced the ability for CRY release, likely due to a possible interaction between SPC and CRY that affects the membrane properties, as confirmed by physical measures
Capsicum annuum L. var. Cornetto di Pontecorvo PDO. Polyphenolic profile and in vitro biological activities
In present study, the potential functional properties of the ethanolic extracts from peel, pulp, edible part and seeds of Capsicum annuum L. var. Cornetto di Pontecorvo were studied. The antimutagenicity against environmental pollutants and oxidative mutagens, the digestive enzyme inhibition, and the antioxidant properties were evaluated in relation to phenolic content. All the samples produced antimutagenic effects against 1-nitropyrene, 2-aminoantracene and tert-butyl-hydroperoxide, and inhibited α-amylase, without affecting α-glucosidase. Also, radical scavenging effects, inhibition of ROS-mediated lipoperoxidation and chelating activity were highlighted. The peel and seed extracts were in general the most effective samples. The phytochemical analysis highlighted a high amount of polyphenols, with a very low carotenoid content. Among phenolics, rutin, gallic acid, carvacrol, chlorogenic acid, 4-OH benzoic acid, and traces of quercetin and epicatechin were determined. Present results highlight a functional role of this pepper ecotype and suggest a possible use of its derived products as food supplements
Chemosensitization of hepatocellular carcinoma cells to sorafenib by β-caryophyllene oxide-induced inhibition of ABC export pumps
Several ATP-binding cassette (ABC) proteins reduce intracellular concentrations of antitumor drugs and hence weaken the response of cancer cells to chemotherapy. Accordingly, the inhibition of these export pumps constitutes a promising strategy to chemosensitize highly chemoresistant tumors, such as hepatocellular carcinoma (HCC). Here, we have investigated the ability of β-caryophyllene oxide (CRYO), a naturally occurring sesquiterpene component of many essential oils, to inhibit, at non-toxic doses, ABC pumps and improve the response of HCC cells to sorafenib. First, we have obtained a clonal subline (Alexander/R) derived from human hepatoma cells with enhanced multidrug resistance (MDR) associated to up-regulation (mRNA and protein) of MRP1 and MRP2. Analysis of fluorescent substrates export (flow cytometry) revealed that CRYO did not affect the efflux of fluorescein (MRP3, MRP4 and MRP5) but inhibited that of rhodamine 123 (MDR1) and calcein (MRP1 and MRP2). This ability was higher for CRYO than for other sesquiterpenes assayed. CRYO also inhibited sorafenib efflux, increased its intracellular accumulation (HPLC–MS/MS) and enhanced its cytotoxic response (MTT). For comparison, the effect of known ABC pumps inhibitors was also determined. They induced strong (diclofenac on MRPs), modest (verapamil on MDR1) or null (fumitremorgin C on BCRP) effect on sorafenib efflux and cytotoxicity. In the mouse xenograft model, the response to sorafenib treatment of subcutaneous tumors generated by mouse hepatoma Hepa 1–6/R cells, with marked MDR phenotype, was significantly enhanced by CRYO co-administration. In conclusion, at non-toxic dose, CRYO is able to chemosensitizating liver cancer cells to sorafenib by favoring its intracellular accumulation