Zinc citrate-coated whey protein nanoparticles alleviate kidney damage and the disturbances in inflammatory gene expression in rats.

This work was conducted to synthesize whey protein nanoparticles (WPNPs) for the coating of zinc citrate (Zn CITR) at three levels and to study their protective role against CCl4 -induced kidney damage and inflammatory gene expression disorder in rats. Seventy male Sprague-Dawley rats were divided into seven groups and treated orally for 4 weeks as follows; the control group, the group treated twice a week with CCl4 (5 mL/kg b.w), the groups received CCl4 plus WPNPs (300 mg/kg b.w); the group received 50 mg/kg b.w of Zn CITR or the three formulas of Zn CITR-WPNPs at low, medium and high doses (LD, MD, and HD). Blood and kidney samples were collected for different assays and histological analyses. The fabricated particles were semispherical, with an average size of 160 ± 2.7, 180 ± 3.1, and 200 ± 2.6 nm and ζ potential of -126, -93, and -84 mV for ZN CITR-WPNPs (LD), Zn CITR-WPNPs (MD), and ZN CITR-WPNPs (HD), respectively. CCl4 significantly increased (p ≤ 0.05) kidney function indices, oxidative stress markers, messenger RNA expression of transforming growth factor-ß1, interleukin (IL)-1ß, IL-10, IL-6, inducible nitric oxide synthase, and tumor necrosis factor-α and significantly decreased (p ≤ 0.05) renal superoxide dismutase, catalase, and glutathione peroxidase along with the histological changes in the kidney tissues. WPNPs, Zn CITR, and Zn CITR loaded WPNPS showed a protective effect against these complications and Zn CITR-WPNPs (LD) was more effective. WPNPs can be used effectively for coating Zn CITR at a level of 7 mg/g WPNPs to be used as a supplement for the protection of the kidney against different toxicants to enhance immunity and avoid harm of excess Zn.

The suppressive role of nanoencapsulated chia oil against DMBA-induced breast cancer through oxidative stress repression and tumor genes expression modulation in rats.

BACKGROUND: Chia oil is high in omega-3 fatty acids, which have been linked to a lower risk of many diseases, including cancer. Oil encapsulation is a method that holds promise for maintaining oil content while enhancing solubility and stability. The purpose of this study is to prepare nanoencapsulated Chia oil and investigate its suppressive effects on rat chemically induced breast cancer. METHODS: The oil was extracted from commercial Chia seeds and their fatty acids were analyzed using Gas Chromatography-mass spectrometry (GC/MS). Sodium alginate was used as a loading agent to create the Chia oil nanocapsules. The DPPH assay was used to assess the oil nanocapsules' capacity to scavenge free radicals. Breast cancer induction was done by single dose subcutaneously administration of 80 mg/kg dimethylbenz (a) anthracene (DMBA). Models of breast cancer were given Chia oil nanocapsules orally for one month at doses of 100 and 200 mg/kg. Through measuring intracellular reactive oxygen species (ROS) and protein carbonyl, assessing the gene expression of tumor suppressor genes (BRCA 1 & 2, TP53), and conducting histopathological analysis, the suppressive effect of Chia oil nanocapsules was examined. RESULTS: The increase in ROS and PC levels brought on by DMBA was significantly decreased by the administration of Chia oil nanocapsules. In tumor tissue from rats given Chia oil nanocapsules, the mRNA expression levels of BRCA1, BRCA2, and TP53 were controlled Histopathological analysis clarified that the tissue architecture of breast tumors was improved by nanocapsules management. CONCLUSIONS: These findings demonstrate the ability of Chia oil nanocapsules to inhibit cancer cells in the rat breast.

Costus speciosus extract protects against the oxidative damage of zearalenone via modulation of inflammatory cytokines, Nrf2 and iNOS gene expression in rats.

Zearalenone (ZEN) is a non-steroidal estrogenic mycotoxin that induces severe health disturbances in humans and animals. This study aimed to determine the bioactive compounds in Costus speciosus extract (CSE) using GC-MS and evaluate its protective capability against ZEN-induced oxidative damage, genotoxicity, and cytotoxicity in rats. Six groups of male Sprague Dawley rats were treated orally for 15 days including the control group, CSE-treated groups at low (200 mg/kg b. w) or high (400 mg/kg b. w) dose, ZEN-treated group (40 µg/kg b. w), and the groups treated with ZEN plus the low or the high dose of CSE. Blood and tissue samples were collected for different assays and pathological analyses. The results of GC-MS indicated the identification of 6 compounds and Azulene was the major. Animals that received ZEN showed severe disturbances in serum biochemical, cytokines, oxidative stress indicators, mRNA expression of iNOS, Nrf2, and inflammatory-related genes. ZEN also increased micronucleated polychromatic erythrocytes (MNPCEs) and comet tail formation in bone marrow cells along with the disturbances in the histological architecture of the liver and kidney. Co-administration of CSE plus ZEN could normalize the majority of the tested parameters and the histological picture at a dose as low as 200 mg/kg b. w. Therefore, CSE protects against ZEN toxicity via its antioxidant activity, modulation of iNOS, inflammatory-related genes, and the Nrf2 pathway and it could be used in the endemic regions.

Nanoencapsulation of basil essential oil alleviates the oxidative stress, genotoxicity and DNA damage in rats exposed to biosynthesized iron nanoparticles.

The application of essential oils in food and pharmaceutical sectors face several challenges due to their sensitivity to oxidation process. Additionally, the biosynthesis of nanometals is growing rapidly; however, the toxicity of these particles against living organisms did not well explore yet. This study aimed to determine the bioactive compounds in basil essential oil (BEO) using GC-MS, to encapsulate and characterize BEO and to evaluate its protective role against the oxidative stress and genotoxicity of biosynthesized iron nanoparticles (Fe-NPs) in rats. Six groups of male Sprague-Dawley rats were treated orally for 4 weeks included the control group, Fe-NPs-treated group (100 mg/kg b.w.); EBEO-treated groups at low (100 mg/kg b.w.) or high (200 mg/kg b.w.) dose and the groups treated with Fe-NPs plus the low or the high dose of EBEO. The GC-MS analysis revealed the identification of 48 compounds and linalool was the major compound. The average sizes and zeta potential of the synthesized Fe-NPs and EBEO were 60 ± 4.76 and 120 ± 3.2 nm and 42.42 mV and -6.4 mV, respectively. Animals treated with Fe-NPs showed significant increase in serum biochemical analysis, oxidative stress markers, cytokines, lipid profile, DNA fragmentation and antioxidant enzymes and their gene expression and severe changes in the histology of liver and kidney tissues. Administration of Fe-NPs plus EBEO alleviated these disturbances and the high dose could normalize most of the tested parameters and improved the histology of liver and kidney. It could be concluded that caution should be taken in using the biosynthesized metal nanoparticles in different application. EBEO is a potent candidate to protect against the hazards of metal nanoparticles and can be applied in food and medical applications.

Bioactive phytochemicals from Salvia officinalis attenuate cadmium-induced oxidative damage and genotoxicity in rats.

This study was conducted to identify the bioactive phytochemicals in Salvia officinalis essential oil, to determine the polyphenols in the aqueous extract (SOE), and to evaluate their protective role against cadmium (Cd)-induced oxidative damage and genotoxicity in rats. Six groups of female rats were treated orally for 2 weeks including the control group, CdCl2-treated group, SOE-treated groups at low or high dose (100 and 200 mg/kg b.w), and CdCl2 plus SOE-treated groups at the two doses. The GC-MS analysis identified 39 compounds; the main compounds were 9-octadecenamide, eucalyptol, palmitic acid, and oleic acid. However, the HPLC analysis showed 12 polyphenolic compounds and the majority were coumaric acid, chlorogenic acid, coffeic acid, catechin, vanillin, gallic acid, ellagic acid, and rutin. In the biological study, rats received CdCl2 displayed severe disturbances in liver and kidney indices alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin (Alb), total protein (TP), total bilirubin (T. Bil), direct bilirubin (D. Bil), creatinine, uric acid, and urea, lipid profile, tumor necrosis factor-alpha (TNF-α), alpha-fetoprotein (AFP) and CEA), glutathione (GSH), glutathione peroxidase (GPx), superoxide dismutase (SOD) and catalase (CAT), malondialdehyde (MDA), nitric oxide (NO), gene expressions, DNA fragmentation, and histological alterations in the liver and kidney tissue. SOE showed a potent antioxidant and mitigated these alterations in serum and tissue. Moreover, the high dose succeeded to normalize most of the tested parameters and histological features. It could be concluded that S. officinalis is a promising source for bioactive compounds with therapeutic benefits against environmental toxicants.

Biosynthesis of gold nanoparticles for the treatment of osteoarthritis alone or in combination with Diacerein® in a rat model.

Gold (Au) compounds were used as an effective therapeutic agent for various inflammatory diseases; however, the use of Au compounds becomes limited because of its association with several side effects. Hence, gold nanoparticles (AuNPs) were developed as a new option for the medical proposes. However, the safety evaluation of gold nanoparticles (AuNPs) in osteoarthritis (OA) treatment remains vague. This study aimed to biosynthesize, characterize and evaluate the therapeutic effects of biosynthesized AuNPs and/or Diacerein® (DIA) in experimental OA. OA was induced by a single injection of monosodium iodoacetate (3 mg/joint) in the intra-articular knee of female rats. Normal rats (N-rats) and OA-rats were treated orally for 5 weeks as follow: untreated N-rats; untreated OA-rats; N-rats received DIA (50 mg/kg b.w); N-rats received AuNPs (30 µg/kg b.w.); N-rats received AuNPs plus DIA; OA-rats received DIA; OA-rats received AuNPs, and OA-rats received AuNPs plus DIA. Blood, knee cartilage, liver and kidney samples were collected for biochemical and histological analysis. The synthesized AuNPs were nearly spherical with average size of 20 nm and zeta potential of 33 mV. AuNPs and DIA induced a significant improvement in serum inflammatory cytokines, biochemical parameters, estrogen level, hepatic and renal oxidative markers, hepatic DNA fragmentation, genomic template stability and cartilage joint histology of OA-rats. AuNPs were more effective than DIA and the combined treatment was more effective than the single treatment. It could be concluded that AuNPs are promising for the treatment of OA alone or in combination with DIA.

Elimination of oxidative stress and genotoxicity of biosynthesized titanium dioxide nanoparticles in rats via supplementation with whey protein-coated thyme essential oil.

The green synthesis of metal nanoparticles is growing dramatically; however, the toxicity of these biosynthesized particles against living organisms is not fully explored. Therefore, this study was designed to synthesize and characterize TiO2-NPs, encapsulation and characterization thyme essential oil (ETEO), and determination of the bioactive constituents of ETEO using GC-MS and evaluate their protective role against TiO2-NPs-induced oxidative damage and genotoxicity in rats. Six groups of rats were treated orally for 30 days including the control group, TiO2-NPs (300 mg/kg b.w)-treated group, ETEO at low (50 mg/kg b.w) or high dose (100 mg/kg b.w)-treated groups, and TiO2-NPs plus ETEO at the two doses-treated groups. Blood and tissues were collected for different assays. The GC-MS results indicated the presence of 21 compounds belonging to phenols, terpene derivatives, and heterocyclic compounds. The synthesized TiO2-NPs were 45 nm tetragonal particles with a zeta potential of -27.34 mV; however, ETEO were 119 nm round particles with a zeta potential of -28.33 mV. TiO2-NPs administration disturbs the liver and kidney markers, lipid profile, cytokines, oxidative stress parameters, the apoptotic and antioxidant hepatic mRNA expression, and induced histological alterations in the liver and kidney tissues. ETEO could improve all these parameters in a dose-dependent manner. It could be concluded that ETEO is a promising candidate for the protection against TiO2-NPs and can be applied safely in food applications.

Fabrication, characterization and biological evaluation of silymarin nanoparticles against carbon tetrachloride-induced oxidative stress and genotoxicity in rats.

This study aimed to synthesize silymarin nanoparticles (SILNPs) using chitosan nanoparticles as a delivery system and to evaluate their protective effects against CCl4 in rats. Eight groups of male Sprague-Dawley rats were treated for three weeks included the control group, CCl4-treated group (100 mg/kg b.w twice a week); SIL-treated group (50 mg/lg b.w); the groups treated daily with low dose (LD) or high dose (HD) of SILNPs (25, 50 mg/kg b.w) and the groups treated with CCl4 plus SIL, SILNPs (LD) or SILNPs (HD). Blood and tissue samples were collected for different assays. The synthesized SILNPs showed a smooth rounded shape with average particle size of 100 ± 2.8 nm. SILNPs contain the same compounds found in raw SIL and the in vitro release of SILNPs continues till 24 h. The in vivo study revealed that SIL and SILNPs at the low or high dose induced a significant improvement in the hematological parameters, liver and kidney function, lipid profile, serum cytokines, gene expression DNA fragmentation and histology of liver and kidney tissue resulted from CCl4. It could be concluded that SILNPs can be applied in oral delivery formulations with a potential application value for liver disease therapy.

Beneficial effects of antioxidants in improving health conditions of sheep infected with foot-and-mouth disease.

The effect of multinutrient antioxidant treatment on sheep naturally infected with FMD virus was investigated in terms of general health conditions, serum proteins profile, and antioxidant/oxidant parameters. Twenty diseased sheep were divided into 4 equal groups (n = 5) and underwent certain therapeutic protocols for 8 weeks as follows: GI, infected not treated group; GII, infected and treated with the ideal and usual line of treatment against FMD virus infection; GIII, infected animals supplemented orally zinc methionine at a dose of 5 g/head/day and vitamin E with selenium-enriched yeast at the same dose level; GIV, infected animals received both the ideal treatment and antioxidants. The animals under experiment were clinically evaluated. Blood samples were obtained for the comet assay and biochemical examination at zero time and at the 8th week after treatment. Results revealed that DNA damage reduced in both GIII and GIV groups which received antioxidants. In the GI group, the activity of SOD and GPx and the level of total antioxidant capacity (TAC) markedly decreased. However, in both GIII and GIV groups treated with multinutrient antioxidants, GPx and TAC values significantly increased after treatment in comparison with the values of the same groups before treatment. After treatment with multinutrient antioxidants, α1-, ß-, and γ-globulins levels markedly increased in GII and GIII groups while α2-globulin level decreased. The improvement in healing of clinical signs and general health conditions was clear in the GIV group. Finally, FMD infection in sheep was found to be associated with oxidative stress. The use of antioxidants as therapeutic approaches recovers and improves general health conditions and performance of affected animals.

Zinc citrate incorporation with whey protein nanoparticles alleviate the oxidative stress complication and modulate gene expression in the liver of rats.

This study aimed to evaluate the hepatoprotective effect of whey protein nanoparticles (WP-NPs) coated Zinc citrate (Zn) against oxidative stress complications and disturbances in gene expression in rats treated with CCl4. WP-NPs were used to coat Zn at three levels and amino acids content was determined in WP-NPs and the fabrications. Seven groups of male albino rats included the control group, CCl4-treated group (0.5 ml/100 g b.w) and the groups treated with CCl4 plus WP-NPs, Zn and the three Zn-WP-NPs fabrications. Blood and liver samples were collected for different analysis. Particles sizes were 95, 142, 196 and 228 nm and zeta potential values were -95, -114, -85 and -79 for WP-NPs and the three Zn-WP-NPs fabrications, respectively. Twelve amino acids were found in WP-NPs and this number was decreased by increasing Zn content. WP-NPs, Zn and the Zn coated WP-NPs counteracted the disturbances in biochemical, parameters, gene expression and histological changes in CCl4-treated rats and Zn-WP-NPs was more effective at the low dose. It could be concluded that WP-NPs enhance the effect of Zn and can be used for coating Zn in the preparation of Zn supplementation to enhance its effect and counteract the side effect of excess Zn.

Modulation of DNA damage and alteration of gene expression during aflatoxicosis via dietary supplementation of Spirulina (Arthrospira) and Whey protein concentrate.

Spirulina (SPN) and Whey protein (WPC) are being touted as functional foods with a number of health benefits. SPN is blue green algae while WPC is a protein complex derived from milk and both have strong antioxidant activity and provoke a free radical scavenging enzyme system. The aim of the present study was to evaluate the antioxidant potentials of SPN and WPC to regulate the alteration of genes' expression and counteract oxidative stress in rats during aflatoxecosis. Eighty male Sprague-Dawley rats were divided into eight groups, which included the control group, the group fed with aflatoxins (AFs)-contaminated diet (2.5 mg/kg diet) for 30 day, the group treated orally with WPC (300 mg/kg b.w.), the group treated orally with SPN (50 mg/kg b.w), the group treated orally with WPC plus SPN and the groups fed with AFs-contaminated diet and treated orally with WPC, SPN and/or WPC. Oxidative stress markers and gene expression were assayed in liver and testis and the damage of DNA was evaluated by DNA fragmentation and micronucleus tests. The results demonstrated that supplementation of SPN and/or WPC reduced the oxidative stress induced by AFs as indicated by decreased lipid peroxidation level, increased glutathione content and up-regulated PHGPx gene expression. Both agents succeed to inhibit DNA damage as indicated by the down-regulation of Fas gene expression, and decreased the percentage of DNA fragmentation and micronucleated erythrocytes. Moreover, WPC was found to be effective than SPN and the combined treatment was more effective than the single treatment. It could be concluded that both SPN and WPC induced a protective action and regulated the alteration of genes expression induced by AFs; however, the combined treatment may be useful than the single treatment.

Dietary supplementation with whey protein and ginseng extract counteracts oxidative stress and DNA damage in rats fed an aflatoxin-contaminated diet.

Aflatoxins (AF) are among the most potent naturally occurring carcinogens and aflatoxin-B1 (AFB(1)) is classified as a group-1 carcinogen. Since the ingestion of aflatoxins-contaminated food is associated with several liver diseases, the aim of the present study was to evaluate whether AF-induced damage in rats can be counteracted by feeding with whey-protein concentrates (WPC) and Korean ginseng extract (KGE). Eighty male Sprague-Dawley rats were divided into eight equal groups and treated daily for 30 days as follows: a control group (fed an AF-free diet), a group fed ad libitum an AF-contaminated diet (2.5mg/kg diet), a group treated orally with WPC (0.5ml/rat/day), a group treated orally with KGE (20mg/kg body weight), a group treated orally with WPC+KGE, and three groups that were fed the AF-contaminated diet and were treated orally with WPC, KGE or WPC+KGE, respectively. Throughout the experimental period, animals received WPC or KGE during the consumption of their respective diet. Bone-marrow micronucleus formation, DNA fragmentation, fatty-acid synthesis (FAS) and phospholipid-hydroperoxide-glutathione-peroxidase (PHGPx) mRNA expression, and oxidative stress were assayed in liver and testis. The results indicated that ingestion of aflatoxin resulted in a significant increase in micronucleated normochromatic erythrocytes (Mn-NCE) in bone marrow, DNA fragmentation, FAS mRNA expression and lipid peroxidation in both organs, and a significant decrease in micronucleated polychromatic erythrocytes/micronucleated normochromatic erythrocytes (PCE/NCE) ratio in bone marrow, PHGPx gene expression and GSH in liver and testis. Treatments with WPC and/or KGE had a significant effect on Mn-NCE or the PCE/NCE ratio in bone marrow. However, KGE or KGE+WPC increased PHGPx gene expression and GSH in testis accompanied with a significant decrease in lipid peroxidation in liver and testis and FAS-mRNA expression in liver. WPC, KGE or WPC+KGE treatments combined with exposure to an AF-contaminated diet restored all the test parameters towards control values, although they did not fully reverse the effects of the aflatoxins. It is suggested that the genotoxicity of aflatoxins can be in part prevented by dietary supplementation with WPC, KGE or their combination.

Aquilegia vulgaris extract protects against the oxidative stress and the mutagenic effects of cadmium in Balb/c mice.

Cadmium (Cd) is a non-essential element and is a widespread environmental pollutant. Exposure to cadmium can result in cytotoxic, carcinogenic and mutagenic effects. The aim of the current work was to evaluate the protective effect of Aquilegia vulgaris extract against the oxidative stress and the genotoxicity induced by Cd using the chromosomal aberrations in somatic and germ cells assay and random amplified polymorphism DNA (RAPD-PCR) analysis. Forty male Balb/c mice were divided into four groups including the control group, Cd-treated group and the groups treated with the extract alone or plus Cd. The results indicated that Cd increased serum ALT, AST, urea, LDH, CK, lipid peroxidation in liver tissue accompanied with a significant decrease in GPX and SOD. Cd also increased the number of chromosomal aberrations in bone marrow and spermatocytes including structural and numerical aberrations. Animals treated with the extract alone were comparable to the control regarding all the tested parameters. The extract succeeded in preventing or diminishing the oxidative stress and the clastogenic effects of Cd. It could be concluded that Aquilegia vulgaris extract is a promising protective agent against oxidative stress and genotoxicity during the exposure to Cd.