Inflammation Factors and Genistein Supplementation in Cancer-Preliminary Research.

The purpose of this study was to evaluate the effect of genistein in nano, micro, and macro forms on the intensity of the DMBA-induced tumor process in rats and to understand the mechanisms of this action. The effect of genistein supplementation on the content of selected eicosanoids (HETEs, HODE, and HEPE) in the serum of rats was evaluated. The levels and expression of genes encoding various pro-inflammatory cytokines (IL-1, IL-6) and MMP-9 in the blood of rats were also investigated. The biological material for the study was blood obtained from female rats of the Sprague Dawley strain (n = 32). The animals were randomly divided into four groups: animals without supplementation, and animals supplemented at a dose of 0.2 mg/kg b.w. (0.1 mg/mL) with macro, micro (587 ± 83 nm), or nano (92 ± 41 nm) genistein. To induce mammary neoplasia (adenocarcinoma), rats were given 7,12-dimethyl-1,2-benz[a]anthracene (DMBA). The content of selected eicosanoids was determined by liquid chromatography with UV detection. An immunoenzymatic method was used to determine the content of cytokines and MMP-9. The expression of the IL-6, IL-1beta, and MMP-9 genes was determined with quantitative real-time PCR (qRT-PCR) using TaqMan probes. Based on the study, it was shown that supplementation of animals with genistein in macro, micro, and nano forms increased the intensity of the tumor process in rats. It was shown that the content of 12-HEPE, HODE, and 12-HETE in the serum of genistein-supplemented rats was statistically significantly lower with respect to the content of the aforementioned markers in the serum of rats receiving only a standard diet, devoid of supplementation. It was found that animals supplemented with nano-, micro-, and macrogenistein had higher levels of metalloproteinase-9, MMP-9, compared to animals without supplementation. There was a significant increase in MMP-9 gene expression in the blood of macrogenistein-supplemented animals, relative to the other groups of rats. On the basis of the study, it was shown that supplementation of animals with nano-, micro-, and macrogenistein had an effect on the development of the tumor process. Dietary supplementation with genistein significantly decreased the level of selected eicosanoids, which may have significant impacts on cancer development and progression.

The Influence of Supplementation with Zinc in Micro and Nano Forms on the Metabolism of Fatty Acids in Livers of Rats with Breast Cancer.

The aim of this study was to investigate the effect of zinc supplementation (in the form of nano or microparticles) on the profile and metabolism of fatty acids in the liver microsomes of rats with induced breast cancer. The activity of desaturases (Δ5, Δ6, Δ9) and the level of cholesterol and its oxidized derivatives were measured. The aim of this study was also to determine the effect of various forms of zinc supplements on rats that were on 5-, 12- and 15-hydroxyeicosatetraenoic (5-, 12- and 15-HETE) and hydroxyoctadecadienoic (HODE) acids, and the level of prostaglandin E2 (PGE2). Female Spraque-Dawley rats (n = 24) were divided into 2 groups that were supplemented with zinc in the micro form (342 nm) or nano form (99 nm) particles, respectively, and a group with a standard diet (control group). All animals received 7,12-dimethylbenz[a]anthracene twice for the induction of breast cancer. Dietary nano-Zn supplementation increased vaccenic acid content (p = 0.032) and decreased Δ6-desaturase activity (p = 0.006), whereas micro-Zn increased cholesterol (p = 0.006), ∑COPs (total cholesterol-oxidation products) (p = 0.019) and PGE2 (p = 0.028) content. Dietary enrichment with Zn microparticles resulted in lower concentrations of the metabolites 15-, 12- and 5-HETE and HODE. Our study indicates that the effect of zinc supplementation on the metabolism of fatty acids in the liver microsomes under neoplastic conditions depends on the form in which it is administered.

Oils' Impact on Comprehensive Fatty Acid Analysis and Their Metabolites in Rats.

Fatty acids, especially polyunsaturated, and their metabolites (eicosanoids) play many pivotal roles in human body, influencing various physiological and pathological processes. The aim of the study was to evaluate the effect of supplementation with edible oils diverse in terms of fatty acid composition on fatty acid contents, activities of converting their enzymes, and on lipoxygenase metabolites of arachidonic and linoleic acids (eicosanoids) in rat serum. Female Sprague-Dawley rats divided into seven groups were used in the study. Animals from six groups were fed one of oils daily (carotino oil, made up by combining of red palm oil and canola oil, linseed oil, olive oil, rice oil, sesame oil, or sunflower oil). One group received a standard diet only. Fatty acids were determined using gas chromatography with flame ionization detection. Eicosanoids-hydroxyeicosatetraenoic (HETE) and hydroxyoctadecadienoic acids (HODE) were extracted using a solid-phase extraction method and analyzed with HPLC. Vegetable oils given daily to rats caused significant changes in serum fatty acid profile and eicosanoid concentrations. Significant differences were also found in desaturases' activity, with the linseed and olive oil supplemented groups characterized by the highest D6D and D5D activity. These findings may play a significant role in various pathological states.

The Effect of Diet Supplementation with Pomegranate and Bitter Melon on Lipidomic Profile of Serum and Cancerous Tissues of Rats with Mammary Tumours.

The aim of this study was to present overall lipid profile of organisms with ongoing neoplastic process and applied diet supplementation with pomegranate seed oil (PSO) and bitter melon extract (BME). The following were quantified in serum and cancerous tissues of rats suffering from mammary tumours: fatty acids, conjugated fatty acids and sterols, their oxidised metabolites (malondialdehyde and oxysterols) and lipoxygenase (LOX) metabolites of polyunsaturated fatty acids. The obtained results indicate that abnormalities in lipid metabolism accompany neoplastic process. These differences concern all classes of lipids and most pathways of their transformation, with the special emphasis on lipid peroxidation and LOX-mediated metabolism. Cancer process appears to be so detrimental that it may conceal positive influence of dietary modifications. The lack of anticarcinogenic properties of PSO and BME in this model may be due to their antioxidant properties or elevated levels of conjugated linoleic acids (CLA), which change CLA isomer activity from anti- to pro-tumorigenic. As CLA are the product of conjugated linolenic acids (CLnA) endogenous metabolism, high CLA levels may be explained by applied diet enrichment.

Inflammation factors and element supplementation in cancer.

The aim of the study was to evaluate the effect of dietary supplementation with chosen minerals (Zn, Se, Fe) on expression of selected cytokines (IL-1, IL-6, TNFα) in spleen of rats and on their concentrations in rat serum under inflammatory and pathological conditions obtained by implantation of prostate cancer cells (LnCaP). Serum levels of metabolites of arachidonic, eicosapentaenoic and linoleic acids (hydroxyeicosatetraenoic, hydroxyeicosapentaenoic and hydroxyoctadecadienoic acids, respectively), as compounds involved in inflammation and cancer development, were also investigated. Male rats were randomised into dietary groups supplemented with Zn, Se or Fe. Prostate cancer cells were implanted to some rats in each group. The study demonstrated that minerals supplemented with the diet may exert various effects on an organism. Selenium, zinc and iron influence pro-inflammatory cytokine expression, what leads to stimulation of inflammation. They also affect synthesis of arachidonic and linoleic acid metabolites that exert pro-inflammatory action and enable cancer development and metastasis.

Impact of conjugated linoleic acid administered to rats prior and after carcinogenic agent on arachidonic and linoleic acid metabolites in serum and tumors.

The objective of the study was to assess the influence of conjugated linoleic acid (CLA) daily supplementation prior and after carcinogenic agent on the concentrations of eicosanoids - metabolites of arachidonic acid (15-, 12- or 5-hydroxyeicosatetraenoic acids (15-, 12-, 5-HETE), prostaglandin E2 (PGE2)) and linoleic acid (13- or 9-hydroxyoctadecadienoic acids (13-, 9-HODE)) in rat serum and 7,12-dimethylbenz[a]anthracene (DMBA)-induced tumors. Female rats were randomised into six groups, receiving 1% or 2% Bio-C.L.A or plant oil since the 37th day of life throughout the whole experiment. Some rats (50-day-old) were administered DMBA to induce tumors. Eicosanoids were analyzed with LC-MS/MS. The study indicated that CLA supplemented daily to rats prior and after carcinogen administration affected concentrations of arachidonic and linoleic acid metabolites in rat serum and induced tumors. However, ratios of eicosanoids exerting opposite activities (e.g. 12-HETE/15-HETE) appear to act as more precise factors reflecting pathological changes in an organism than individual compounds.

Influence of pomegranate seed oil and bitter melon aqueous extract on polyunsaturated fatty acids and their lipoxygenase metabolites concentration in serum of rats.

Competition with polyunsaturated fatty acids (PUFA) and an impact on eicosanoid biosynthesis may be one of mechanisms of conjugated linolenic acids (CLnA) action. The aim of this study was to investigate the influence of diet supplementation with pomegranate seed oil, containing punicic acid (PA)-one of CLnA isomers, and an aqueous extract of dried bitter melon fruits, administered separately or together, on PUFA and their lipoxygenase metabolites' concentration in serum of rats. Percentage share of fatty acids was diversified in relation to applied supplementation. PA was only detected in serum of pomegranate seed oil supplemented group, where it was about 1%. Cis-9, trans-11 conjugated linoleic acid (rumenic acid, RA) level tended to increase in group supplemented simultaneously with both dietary supplements whereas its highest share in total fatty acids pool was detected in group receiving solely bitter melon dried fruits aqueous extract. This indicates that consumption of bitter melon tea significantly increased RA content in fatty acids pool in serum. However, pomegranate seed oil elevated procarcinogenic 12-hydroxyeicosatetraenoic acid concentration. Taking into account that pomegranate seed oil and bitter melon dried fruits are dietary supplements accessible worldwide and willingly consumed, the biological significance of this phenomenon should be further investigated. We presume, that there may be a need for some precautions concerning the simultaneous use of these products.

Influence of maternal diet enrichment with conjugated linoleic acids on lipoxygenase metabolites of polyunsaturated fatty acids in serum of their offspring with 7,12-dimethylbenz[a]anthracene induced mammary tumors.

Conjugated linoleic acids (CLA), which are a group of naturally occurring in food isomers of linoleic acid, seem to be active in each step of cancer development. There are many possible mechanisms of this action, and interactions with polyunsaturated fatty acids (PUFA) in lipoxygenase (LOX) and cyclooxygenase (COX) pathways are among the most likely ones. The aim of this study was to assess the influence of diet supplementation with CLA of pregnant and breastfeeding Sprague-Dawley female rats on selected polyunsaturated fatty acids and their LOX metabolites concentrations in serum of the progeny with chemically induced mammary tumors. We confirmed that higher supply of CLA in the diet of female rats corresponded with the lower susceptibility to chemically induced mammary tumors in their female offspring. It also influenced the polyunsaturated n-3 and n-6 fatty acid concentrations in serum, as well as the concentrations of their LOX metabolites. The significant negative correlation between the concentrations of two CLA isomers in serum and linoleic acid (p=0.0144, p=0.0098), eicosapentaenoic acid (p=0.0158, p=0.0124), and 5-HEPE (p=0.0014, p=0.01690) and between cis-9, trans-11 CLA and 15-HEPE was detected, whereas arachidonic acid concentration positively correlated with CLA concentration in serum (p=0.0150, p=0.0231). Our results indicate that CLA can compete with PUFA and influence serum concentration of PUFA and their LOX metabolites, which could partly explain the anticancerogenic action of CLA.

Effect of conjugated linoleic acid mixture supplemented daily after carcinogen application on linoleic and arachidonic acid metabolites in rat serum and induced tumours.

Conjugated linoleic acid (CLA) is thought to exert anticarcinogenic, antiatherogenic, anti-inflammatory and weight loss effects. The impact on eicosanoid biosynthesis may be one of the mechanisms of its action. The aim of this study was to establish whether CLA mixture supplemented daily after administration of carcinogen (7, 12-dimethylbenz[a]anthracene, DMBA) influenced the concentration of linoleic and arachidonic acid metabolites: 13- or 9-hydroxyoctadecadienoic acids (13-, 9-HODE) and 15-, 12- or 5-hydroxyeicosatetraenoic acids (15-, 12- or 5-HETE) and prostaglandin E2 (PGE2) in rat serum and DMBA-induced tumours. The correlations between polyunsaturated fatty acids (PUFA) and HETE and HODE contents in serum were also investigated. Female Sprague-Dawley rats divided into three groups according to the diet (1% Bio-C.L.A., 2% Bio-C.L.A. and plant oil in the control group) were used in the study. On the 50th day of life some of the animals in every dietary group were administered DMBA to induce tumours. Since that day, the rats were fed one of the above-mentioned diets. After 15weeks the animals were sacrificed and blood and tumours were collected. HETE and HODE were extracted using a solid-phase extraction (SPE) method on C18 columns and analysed with LC-MS/MS. The results of our study showed that CLA daily supplementation after carcinogen administration influence LA and AA metabolite levels in serum and tumours. However, the ratios of eicosanoids having opposite effects (e.g. 12-HETE/15-HETE), not concentrations of particular compounds, appear to be better indicators of pathological processes.

Effects of dietary linseed, evening primrose or fish oils on fatty acid and prostaglandin E2 contents in the rat livers and 7,12-dimethylbenz[a]anthracene-induced tumours.

We examined the influence of diets supplemented with fish and vegetable oils on fatty acid and prostaglandin E2 (PGE2) contents in livers of non-7,12-dimethylbenz[a]anthracene (DMBA)- and DMBA-treated rats, and in DMBA-induced tumours. Decreased concentrations of saturated fatty acids and increased unsaturated fatty acid levels were observed in liver phospholipids of rats fed these oils. There was a marked difference in the concentrations of fatty acids found in the tumours and those present in liver lipids. Oleic acid was the main unsaturated fatty acid found in the tumour tissue. Both liver and tumour PGE2 contents were clearly correlated to the diet. The PGE2 concentrations were decreased in livers and tumours of rats fed fish (FO) and linseed oils (LO).