Simvastatin Reduces Doxorubicin-Induced Cardiotoxicity: Effects beyond Its Antioxidant Activity.

This study aimed to evaluate if Simvastatin can reduce, and/or prevent, Doxorubicin (Doxo)-induced cardiotoxicity. H9c2 cells were treated with Simvastatin (10 µM) for 4 h and then Doxo (1 µM) was added, and the effects on oxidative stress, calcium homeostasis, and apoptosis were evaluated after 20 h. Furthermore, we evaluated the effects of Simvastatin and Doxo co-treatment on Connexin 43 (Cx43) expression and localization, since this transmembrane protein forming gap junctions is widely involved in cardioprotection. Cytofluorimetric analysis showed that Simvastatin co-treatment significantly reduced Doxo-induced cytosolic and mitochondrial ROS overproduction, apoptosis, and cytochrome c release. Spectrofluorimetric analysis performed by means of Fura2 showed that Simvastatin co-treatment reduced calcium levels stored in mitochondria and restored cytosolic calcium storage. Western blot, immunofluorescence, and cytofluorimetric analyses showed that Simvastatin co-treatment significantly reduced Doxo-induced mitochondrial Cx43 over-expression and significantly increased the membrane levels of Cx43 phosphorylated on Ser368. We hypothesized that the reduced expression of mitochondrial Cx43 could justify the reduced levels of calcium stored in mitochondria and the consequent induction of apoptosis observed in Simvastatin co-treated cells. Moreover, the increased membrane levels of Cx43 phosphorylated on Ser368, which is responsible for the closed conformational state of the gap junction, let us to hypothesize that Simvastatin leads to cell-to-cell communication interruption to block the propagation of Doxo-induced harmful stimuli. Based on these results, we can conclude that Simvastatin could be a good adjuvant in Doxo anticancer therapy. Indeed, we confirmed its antioxidant and antiapoptotic activity, and, above all, we highlighted that Simvastatin interferes with expression and cellular localization of Cx43 that is widely involved in cardioprotection.

Evaluation of the Antioxidant Activity of Nigella sativa L. and Allium ursinum Extracts in a Cellular Model of Doxorubicin-Induced Cardiotoxicity.

Natural products black cumin-Nigella sativa (N. sativa) and wild garlic-Allium ursinum (AU) are known for their potential role in reducing cardiovascular risk factors, including antracycline chemotherapy. Therefore, this study investigates the effect of N. sativa and AU water and methanolic extracts in a cellular model of doxorubicin (doxo)-induced cardiotoxicity. The extracts were characterized using Ultraviolet-visible (UV-VIS) spectroscopy, Fourier-transform infrared (FT-IR) spectroscopy, Liquid Chromatography coupled with Mass Spectrometry (LC-MS) and Gas Chromatography coupled with Mass Spectrometry (GC-MS) techniques. Antioxidant activity was evaluated on H9c2 cells. Cytosolic and mitochondrial reactive oxygen species (ROS) release was evaluated using 2',7'-dichlorofluorescin-diacetate (DHCF-DA) and mitochondria-targeted superoxide indicator (MitoSOX red), respectively. Mitochondrial membrane depolarization was evaluated by flow cytometry. LC-MS analysis identified 12 and 10 phenolic compounds in NSS and AU extracts, respectively, with flavonols as predominant compounds. FT-IR analysis identified the presence of carbohydrates, amino acids and lipids in both plants. GC-MS identified the sulfur compounds in the AU water extract. N. sativa seeds (NSS) methanolic extract had the highest antioxidant activity reducing both intracellular and mitochondrial ROS release. All extracts (excepting AU methanolic extract) preserved H9c2 cells viability. None of the investigated plants affected the mitochondrial membrane depolarization. N. sativa and AU are important sources of bioactive compounds with increased antioxidant activities, requiring different extraction solvents to obtain the pharmacological effects.

Nigella sativa: Valuable perspective in the management of chronic diseases.

OBJECTIVES: Over the past 20 years, increasing interest in the use of medicinal plants as alternative or adjuvant treatments of several chronic diseases was observed. Accordingly, Nigella sativa or black cumin, a medicinal plant rich in bioactive compounds, has been used worldwide for food purposes or in traditional medicines. This paper aims to reveal N. sativa potential as adjunct treatment in cardiovascular diseases, diabetes, and hematological malignancies, due to their increasing prevalence and difficult management in everyday life. MATERIALS AND METHODS: Databases like PubMed, Web of Science, Science Direct, Scopus, and Google Scholar were used to search the literature data. Keywords like anti-inflammatory effect, anti-oxidant effect, antihypertensive effects, hypolipidemic effects and hematological malignancies were used in combination with N. sativa. RESULTS: Because of its numerous pharmacological actions, but especially for its anti-oxidant and anti-inflammatory properties, in vivo and in vitro studies demonstrated N. sativa positive effect against diabetes, hypertension, and hypercholesterolemia, all of them associated to cardiovascular diseases progression. Also, it was proved to have marked anti-proliferative, cytotoxic, pro-apoptotic, and anti-metastatic effects, in both solid cancers and hematological malignancies. CONCLUSION: N. sativa used as complementary treatment to classical medications can improve the management of several chronic diseases.

Evaluation of heavy metals, cytotoxicity, and antioxidant activity of tomatoes grown in toxic muddy soils.

This research studies tomatoes grown in polluted soils to ascertain their phytochemical and nutritive features. Pulp and seeds from tomatoes grown in muddy soils were analyzed for their antioxidant power and their toxicity because of the possibility that heavy metals were present in the soils. An antioxidant assay on methanol extracts was made by using DDPH, while an ABTS [2,2'-Azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid)] assay was used to evaluate the antioxidant activity of lipophilic fractions. Results of the antioxidant assay showed that the tomatoes maintained a high level of antioxidant activity especially in the lipophilic fractions which contain the most representative compounds. Cytotoxic activity was performed on HeLa, PDAC, and A375 cell lines by [3-(4,5-dimethylthiazol-2-yl)-2,5-phenyl-2H-tetrazolium bromide] (MTT) assay. Results showed that neither the seeds, nor the pulp, of the extracts was cytotoxic. The presence of heavy metals was evaluated by using spectroscopy of atomic absorption with a graphite oven. Test results show the absence of heavy metals and these results have an interesting scientific role because they provide useful information for promoting food safety.

L-Arginine and its metabolites in kidney and cardiovascular disease.

L-Arginine is a semi essential amino acid synthesised from glutamine, glutamate and proline via the intestinal-renal axis in humans and most mammals. L-Arginine degradation occurs via multiple pathways initiated by arginase, nitric-oxide synthase, Arg: glycine amidinotransferase, and Arg decarboxylase. These pathways produce nitric oxide, polyamines, proline, glutamate, creatine and agmatine with each having enormous biological importance. Several disease are associated to an L-arginine impaired levels and/or to its metabolites: in particular various L-arginine metabolites may participate in pathogenesis of kidney and cardiovascular disease. L-Arginine and its metabolites may constitute both a marker of pathology progression both the rationale for manipulating L-arginine metabolism as a strategy to ameliorate these disease. A large number of studies have been performed in experimental models of kidney disease with sometimes conflicting results, which underlie the complexity of Arg metabolism and our incomplete knowledge of all the mechanisms involved. Moreover several lines of evidence demonstrate the role of L-arg metabolites in cardiovascular disease and that L-arg administration role in reversing endothelial dysfunction, which is the leading cause of cardiovascular diseases, such as hypertension and atherosclerosis. This review will discuss the implication of the mains L-arginine metabolites and L-arginine-derived guanidine compounds in kidney and cardiovascular disease considering the more recent literature in the field.

Antioxidant and cytotoxic activities investigation of tomato seed extracts.

Biological activities of different varieties of tomato seed extracts were evaluated to verify the potential antioxidant and/or antiproliferative activity of the bioactive metabolites present in them. Findings demonstrated that among all the varieties investigated (San Marzano Rosso, San Marzano Giallo, Corbarino, Black Tomato and San Marzano/Black Tomato hybrid) San Marzano Rosso seed extract exhibited the highest free radical-scavenging activity with 68% of 2,2-Diphenyl-1-picrylhydrazyl radical inhibition, and the best cytotoxic activity evaluated by using the brine shrimp test (LD50: 23,198 ppm) and 3-(4,5-dimethylthiazol-2-yl)-2,5-phenyl-2H-tetrazolium bromide assay on A375 cell line (IC50: 137.7 µg/mL).

Cytotoxic activity of nemorosone in human MCF-7 breast cancer cells.

Estrogen receptor (ER) antagonists have been widely used for breast cancer treatment; however, patients have increasingly shown resistance and sensitivity to the high toxicity of these drugs, and identification of novel targeted therapies is therefore required. To determine whether nemorosone, a polycyclic polyisoprenylated benzophenone isolated from floral resins of Clusia rosea Jacq. and Cuban propolis samples, exerts anticancer effects on human breast cancer cells, estrogen receptor positive (ERα+) MCF-7 and estrogen receptor negative (ERα-) MDA-MB-231 and LNCaP cells were used. Cells were treated with nemorosone alone or in association with 17ß-estradiol (E2) or an ER antagonist, ICI 182,780, a selective ER downregulator that completely abrogates estrogen-sensitive gene transcription. Nemorosone inhibited the cell viability of ERα+ but not of ERα- cells. In MCF-7, nemorosone induced inhibition of cell growth by blocking the cell cycle in the G0/G1 phase. Moreover, the expression of pERK1/2 and pAkt, considered to be hallmarks of the nongenomic estrogen signalling pathway, were reduced in MCF-7 cells treated with nemorosone. All these effects were enhanced by ICI 182,780. However, nemorosone was not able to interfere with E2-induced Ca²(+) release. These findings suggest that nemorosone may have therapeutic application in the treatment of breast cancer because of its activity on ERα.

Antiproliferative activity of brown Cuban propolis extract on human breast cancer cells.

Brown Cuban propolis (BCP) is the major type of propolis in Cuba; its chemical composition is exclusive and the principal component is nemorosone. In this study we investigated the antiproliferative activity of the ethanol extract of BCP on human breast cancer cell lines. The MTT assay showed a significant antiproliferative activity (P<0.005) of BCP on MCF-7 (estrogen receptor positive ER+) rather than MDA-MB 23 1 (ER-). This effect was concentration- (1-25 microg/mL) and time- (24-48 h) dependent, but it is only partially attributable to apoptosis. Indeed, our data showed that BCP administration to MCF-7 caused a significant (P>0.01) inhibition of cell growth in the G1 phase of cell cycle, which was mechanism dose- and time-dependent. 17-beta Estradiol (10 nM) administration to MCF-7 caused a significant (P<0.001), but not total reduction of BCP antiproliferative activity at concentrations of 1, 5 and 10 microg/mL, but not at the highest concentration (25 microg/mL). The coadministration of ICI 182,780 (100 nM), an antagonist of ER, on MCF-7 totally reduced the effect of BCP at 24 h, and showed a significant (P<0.001) reduction of BCP antiproliferative activity at 48 h. Thus it was hypothesized that BCP possesses an estrogen-like activity. It is to be noted, however, that BCP application to MDA-MB 23 1 at 48 h also induced increased cell mortality. Thus, it cannot be ruled out that BCP could not only interact with the ER, but also have an ER-independent activity.