Eucalyptol regulates Nrf2 and NF-kB signaling and alleviates gentamicin-induced kidney injury in rats by downregulating oxidative stress, oxidative DNA damage, inflammation, and apoptosis.

Gentamicin, an aminoglycoside antibiotic, is nowadays widely used in the treatment of gram-negative microorganisms. The antimicrobial, anti-inflammatory, and antioxidant activities of eucalyptol, a type of saturated monoterpene, have been reported in many studies. The aim of this study was to examine the possible effects of eucalyptol on gentamicin-induced renal toxicity. A total of 32 rats were divided into 4 groups; Control (C), Eucalyptol (EUC), Gentamicin (GEN), and Gentamicin + Eucalyptol (GEN + EUC). In order to induce renal toxicity, 100 mg/kg gentamicin was administered intraperitoneally (i.p.) for 10 consecutive days in the GEN and GEN + EUC groups. EUC and GEN + EUC groups were given 100 mg/kg orally of eucalyptol for 10 consecutive days. Afterwards, rats were euthanized and samples were taken and subjected to histopathological, biochemical, immunohistochemical, and real-time PCR examinations. The blood urea nitrogen (BUN) and creatinine (CRE) levels were significantly decreased in the GEN + EUC group (0.76 and 0.69-fold, respectively) compared to the GEN group. The glutathione peroxidase (GPx) and catalase (CAT) activities were significantly increased in the GEN + EUC group (1.35 and 2.67-fold, respectively) compared to the GEN group. In GEN group, Nuclear factor kappa B (NF-kB), Interleukin 1-beta (IL-1ß), Inducible nitric oxide synthase (iNOS), Tumor necrosis factor-α (TNF-α), Caspase-3, 8-Hydroxy-2'-deoxyguanosine (8-OHdG) and Nuclear factor erythroid 2-related factor (Nrf2) expression levels were found to be quite irregular. GEN + EUC group decreased the expressions of NF-kB, IL-1ß, iNOS, TNF-α, Caspase-3, and 8-OHdG (0.55, 0.67, 0.54, 0.54, 0.63 and 0.67-fold, respectively), while it caused increased expression of Nrf2 (3.1 fold). In addition, eucalyptol treatment ameliorated the histopathological changes that occurred with gentamicin. The results of our study show that eucalyptol has anti-inflammatory, antioxidative, antiapoptotic, nephroprotective, and curative effects on gentamicin-induced nephrotoxicity.

Eucalyptol alleviates cisplatin-induced kidney damage in rats.

This study was aimed to explore the therapeutic effect of eucalyptol on cisplatin induced kidney damage in Wistar albino rats. The animals were divided into four groups: sham (S), eucalyptol (E), cisplatin (C), and cisplatin + eucalyptol (CE) randomly, six animals in each group. Groups C and CE were received cisplatin (12 mg/kg, a single dose, intraperitoneally (i.p.)). Groups E and CE were treated with eucalyptol (100 mg/kg, for seven days, orally). The blood samples and kidney tissues were collected following sacrification and analyzed histopathologically and biochemically. Histopathological results revealed tubular degeneration and necrosis, inflammatory cell infiltration, tubular lumen dilatation, enlargement of bowman's space and hyaline cast were significantly irregular in the group C than group S. However, eucalyptol treatment (CE) modulated the alterations in the group C. Serum levels of blood urea nitrogen (BUN) and creatinine (CRE) were considerably higher in the group C compared to the other groups. There was no significant difference among the other groups statistically (except group C) in terms of BUN and CRE values. Eucalyptol treatment (at 100 mg/kg, for seven days) decreased the cisplatin induced increase in serum BUN and CRE levels and restored the reduced Vit C level and CAT activity of kidneys caused by cisplatin. Thus, eucalyptol's antioxidative, nephroprotective, and curative effects indicated the potential for future drug development.

Apigenin alleviates neuroinflammation in a mouse model of Parkinson's disease.

PURPOSE OF THE STUDY: The aim of this study is to evaluate the effect of apigenin on inflammatory response in brain tissue in Parkinson's mouse model. MATERIALS AND METHODS: Parkinson's disease model was induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Sixty 8-10-weeks-old male C57BL/6 mice were randomly divided into four groups control, Parkinson, prophylaxis, and treatment. Control (0.9% NaCl 0.5 ml, 10 days, i.p.), Parkinson (25 mg/kg MPTP, 5 days, i.p.), prophylaxis (50 mg/kg apigenin, 5 days + 25 mg/kg MPTP, 5 days, i.p.), and treatment (25 mg/kg MPTP, 5 days + 50 mg/kg apigenin, 5 days). The expressions and protein levels of tumor necrosis factor-alpha (TNF-α), interleukin-1-beta (IL-1ß), IL-6, IL-10, and transforming growth factor-beta (TGF-ß) were determined using immunohistochemistry and enzyme-linked immunosorbent analysis. RESULTS: Apigenin administration attenuated MPTP-induced histopathological changes in brain tissue. Furthermore, apigenin reversed the changes in expressions and concentrations of TNF-α, IL-1ß, IL-6, IL-10, and TGF-ß. CONCLUSION: This study suggests that apigenin could be used as a neuroprotective option to attenuate neuroinflammation in Parkinson's disease.

Protective effects of nobiletin on cisplatin induced neurotoxicity in rats.

OBJECTIVES: This study was designed to investigate the possible antioxidant, antiapoptotic and neuroprotective effects of nobiletin on cisplatin-induced neurotoxicity rat model by evaluating neurotrophins, antioxidants and histopathology. METHODS: Forty male Wistar Albino rats were divided into four groups: control, cisplatin (CIS), cisplatin + nobiletin (CIS + NOB) and nobiletin + cisplatin (NOB + CIS). CIS + NOB was applied nobiletin (10 mg/kg, i.p.) during the last four days whereas NOB + CIS was applied nobiletin during the first four days of the study. Cisplatin (4 mg/kg, i.p. twice a day) was administered to the experimental groups on the 5th day of the study. All rats were sacrificed on the 10th day of the study. BDNF, NGF, G6PD, GPx, tGSH and MDA levels were determined in brain. In addition, routin histolopathological analysis and caspase-3 immunoreactivity assay were conducted. RESULTS: BDNF concentrations increased in nobiletin-administered groups, compared to Control and CIS and that the increase was statistically significant in NOB + CIS (p < 0.05). It was also found that G6PD activity increased (p < 0.05) in the nobiletin-administered groups, compared to control and CIS. Histopathologically, neuronal degeneration, oedema and gliosis increased in CIS compared to Control, and nobiletin administration decreased neuronal degeneration and oedema compared to CIS (p < 0.05). Cisplatin increased (p < 0.05) caspase-3 immunoreactivity in cerebrovascular endothelium and neurons compared to Control, while nobiletin administration decreased caspase-3 immunoreactivity in cerebrovascular endothelium. Caspase-3 immunoreactivity in neurons decreased only in NOB + CIS (p < 0.05). CONCLUSION: Nobiletin increased BDNF concentration and G6PD activity in brain and when evaluated together with histopathological and immunohistochemical findings, it may have antioxidant, antiapoptotic and neuroprotective effects against cisplatin.

Nobiletin alleviates methotrexate-induced hepatorenal toxicity in rats.

We investigated the possible ameliorative effects of nobiletin (NBL) against methotrexate (MTX)-induced hepatorenal toxicity in rats. Twenty-eight Wistar albino rats were randomly divided into four groups, namely: Control; MTX (administered 20 mg/kg MTX); MTX+NBL (administered 20 mg/kg MTX and 10 mg/kg NBL per day); and NBL (administered 10 mg/kg/day NBL). Histopathological, immunohistochemical and biochemical analyses were performed on the kidney and liver tissues of rats at the end of the study. MTX caused renal toxicity, as indicated by increases in malondialdehyde (MDA) and caspase-3, as well as decreases in reduced glutathione (GSH), glucose-6-phosphate dehydrogenase (G6PD), glutathione peroxidase (GPx), catalase (CAT) and B-cell lymphoma-2 (Bcl-2). MTX also caused hepatotoxicity, as indicated by increases in 8-hydroxy-2'-deoxyguanosine (8-OHdG), tumor necrosis factor alpha (TNF-α), MDA and caspase-3 and decrease in interleukin 10 (IL-10), GSH, total antioxidant capacity, GPx, G6PD, CAT and Bcl-2. MTX caused histopathological changes in kidney and liver tissues indicating tissue and cellular damage. Administration of NBL concurrently with methotrexate reduced oxidative stress, inflammatory and apoptotic signs, and prevented kidney and liver damage caused by methotrexate. We consider NBL has attenuating and ameliorating effects on methotrexate-induced hepatorenal toxicity.

Protective and therapeutic effects of nobiletin against cisplatin-induced nephrotoxicity in rats.

Possible protective and therapeutic effects of nobiletin on kidney in a cisplatin-induced nephrotoxicity rat model were investigated. Forty male albino rats were divided into four groups: control, cisplatin (CIS), cisplatin+nobiletin (CIS+NOB), and nobiletin+cisplatin (NOB+CIS). At the end of the study, the rats were subjected to biochemical, histological and immunohistochemical analyzes. Compared to the control group, tGSH (p < 0.05) levels, and G6PD (p < 0.05) and GPx (p < 0.001) activities, were increased in the CIS group; while significant (p < 0.05) decreases occurred in the MDA and TOC levels. Histopathologically, the kidneys of the groups administered nobiletin (CIS+NOB, NOB+CIS) were significantly different from the CIS group, being closer to control group in terms of degeneration and hyaline cylinder formation in the tubules (p < 0.05). While dilatation in the tubules, protein-rich fluid and hyaline cylinder formation in the lumen were most common in the CIS group, a significant decrease (p < 0.05) of these parameters was seen in the nobiletin groups (CIS+NOB, NOB+CIS). This study suggests that nobiletin can be effective in preventing and ameliorating toxic effects of cisplatin on the kidney.

Proanthocyanidin alleviates testicular torsion/detorsion-induced ischemia/reperfusion injury in rats.

Testicular torsion is an urological emergency and can lead to ischemia damage and testicular loss if not diagnosed in time. Proanthocyanidin is reported to have anti-inflammatory and antioxidant properties. The current study aimed to examine the possible effects of proanthocyanidin (P) on the testis in torsion/detorsion (T/D)-induced testicular ischemia/reperfusion (I/R) injury in rats. Forty rats were divided into four groups (n=10 for each): sham-operated (sham), I/R, I/R + P100 (100 mg/kg, 30 min before torsion), and I/R + P200 (200 mg/kg, 30 min before torsion). Testicular T/D was performed on the left testicle by 3 hours of torsion at 720° clockwise, followed by 3 hours of detorsion. In the I/R group, an increase in malondialdehyde (MDA) levels and a decrease in glutathione (GSH), vitamin C (Vit C), glutathione peroxidase (GPx), glucose-6-phosphate dehydrogenase (G6PD) values were determined compared to the sham group (p<0.001). Moreover, an increase in the expression of cleaved caspase-3 and Bcl2-associated X protein (Bax), a decrease in the expression of B-cell lymphoma 2 (Bcl-2) and proliferating cell nuclear antigen (PCNA) were detected in the I/R group (p<0.001). Histopathologically, it was determined that the Johnsen and Cosentino scores of the testicles were irregular in the I/R group (p<0.001). Proanthocyanidin treatment caused a decrease in MDA, cleaved caspase-3 and Bax levels and an increase in GSH, Vit C, GPx, G6PD, Bcl-2 and PCNA values. Additionally, Johnsen and Cosentino rearranged the scores. The present findings revealed the protective and curative effects of proanthocyanidin in organ damage due to testicular torsion/detorsion-induced ischemia/reperfusion with their antioxidative and antiapoptotic properties.

Effects of black cumin (Nigella sativa L.) seed on growth performance, blood parameters, liver oxidant/anti-oxidant levels and fatty liver syndrome in quails.

This research aimed to evaluate the effect of different doses of black cumin (Nigella sativa L.) seed (BCS) on growth performance, blood parameters, liver oxidant/anti-oxidant levels and fatty liver syndrome in quails. Four hundred and thirty-two unsexed (male and female) three-day-old Japanese quail (Coturnix coturnix japonica) chicks were divided into four treatment groups (108 chicks per group) with six replicates (18 chicks per replicate). Control and experimental groups were fed for 35 days with basal quail feed including 0.00, 0.50, 1.00 and 2.00% BCS supplement, respectively. At the end, a total of 96 quails, 24 from each group (12 females and 12 males) were slaughtered. The BCS-addition did not affect the growth performance in the experimental groups compared to the control group. Addition of BCS to the diet significantly decreased serum aspartate aminotransferase, lactate dehydrogenase and urea amounts compared to the controls. Whereas, cholesterol decreased significantly with the addition of only 1.00% and low-density lipoprotein with the addition of 0.50 and 1.00% BCS compared to the controls. Liver glutathione levels significantly elevated in 0.50 and 1.00% BCS fed groups; while, glutathione peroxidase levels significantly decreased in 1.00 and 2.00% BCS fed groups. Adding 1.00 and 2.00% BCS to the feed reduced fatty liver incidence in male quails. It is concluded that adding 0.50 and 1.00% BCS positively affects the blood and liver parameters; therefore, BCS may be suggested as an anti-oxidant source to help protect hepatocytes against tissue damage as it has a significant effect on maintaining oxidant and anti-oxidant balance.

Epidermal growth factor concentration in milk of healthy water buffaloes (Bubalus bubalis).

Epidermal growth factor (EGF) has biological roles, including embryonic organ development, breast morphogenesis, breast cell proliferation, and mammary development. This study aimed to measure EGF concentration and evaluate its relationship with somatic cell count (SCC) in healthy water buffaloes (Bubalus bubalis) milk. The study material was constituted of 120 milk samples obtained from 30 healthy water buffaloes between the ages of 3 - 6 years, negative for California mastitis test and SCC less than 3.00 × 105 cells mL-1 milk. In milk serum samples, the EGF concentration was measured using a bovine-specific enzyme-linked immunosorbent assay kit. Epidermal growth factor concentration in the buffalo milk was ranged from 4.30 to 9.80 ng mL-1, with a mean of 8.30 ± 1.50 ng mL-1. Positive correlation between milk SCC values and EGF concentrations was recorded in water buffaloes. Further research is required to evaluate the content of milk EGF in different species of animals because of the EGF effective role in mammary gland and intestinal mucosa.

Determination of Serum Oxidative Stress, Antioxidant Capacity and Protein Profiles in Dogs Naturally Infected with Ehrlichia canis.

PURPOSE: Canine ehrlichiosis is an important tick-borne disease of dogs worldwide. In the present study, we aimed to determine the serum total antioxidant capacity (TAC), malondialdehyde (MDA), nitric oxide (NO), superoxide dismutase, (SOD), glutathione peroxidase (GSH-Px), adenosine deaminase (ADA) activity and serum protein profiles in dogs affected with naturally acquired ehrlichiosis. METHODS: The animal materials had been consisted of ten dogs naturally infected with Ehrlichia canis, and ten controls negative for Ehrlichia canis. TAC, MDA, NO, SOD, GSH-Px, ADA activity and TP, ALB, GLOB levels were measured in sera of the animals. The serum protein concentrations were measured by autoanalyzer. The electrophoretic profiles of serum total protein were determined by native polyacrylamide gel electrophoresis (Native-PAGE). RESULTS: In dogs with ehrlichiosis, decreased TAC (P < 0.05) and GSH-Px (P > 0.05) levels were determined. However, NO (P > 0.05), SOD (P < 0.05), ADA (P > 0.05), MDA (P > 0.05), TP (P < 0.05) and GLO (P < 0.05) levels were found as increased in the Ehrlichia positive dogs. ALB levels were decreased without a statistical significance (P > 0.05). ALB, α1 and ß2 globulin strip densities were found as decreased in native-PAGE, while ß1 and γ globulin strip densities were significantly increased in the E. canis positive group when compared to the control. CONCLUSION: It was determined that the oxidative stress decreased high antioxidant activity in dogs naturally infected with E. canis, and consequently, pro-oxidant and antioxidant defense and serum protein profiles were affected. It was thought that antioxidant supplementation could be beneficial to the treatment of the disease.

Determination of Quality Criteria that Allow Differentiation Between Honey Adulterated with Sugar and Pure Honey.

This study used various parameters of honey to develop a potentially more robust approach to the detection of adulterated honey. For this purpose, 25 multifloral, natural honey samples and 20 samples of adulterated honey produced by bees that had been fed supplementary sucrose syrup were analysed. The mean total phenolic content of the natural honeys was considerably higher than in the adulterated honeys at 157 ± 13 and 35.2 ± 7.3 mg GAE/100 g, respectively. Similarly, considerable variation was determined between natural and adulterated honeys in terms of total flavonoids (3.3 ± 0.3 and 2.1 ± 0.4 mg QE/100 g, respectively), antiradical activity (87.9 ± 12 and 163 ± 11 mg/mL, respectively) and proline content (202 ± 26 and 71.1 ± 21.6 mg/kg, respectively.) The potassium, phosphorus, calcium and magnesium contents of natural honeys were also higher than in adulterated honeys (P < 0.01). In conclusion, the determination of the proline level, phenolic content, antioxidant activity and mineral profile may collectively provide a more holistic method approach to the differentiation of natural and adulterated honey, and also for comparing their food values.

Erythrocyte and spermatozoa glucose-6-phosphate dehydrogenase activity in merino rams: An experimental study.

BACKGROUND: Glucose-6-phosphate dehydrogenase (G6PD) is the first enzyme of the pentose phosphate metabolic pathway that supplies reducing agents by maintaining the level of reduced nicotinamide adenine dinucleotide phosphate. OBJECTIVE: It was aimed to determine the activity of erythrocyte and spermatozoa G6PD in the breeding and non-breeding seasons in Merino rams. And also, to find out the relation of these parameters with sperm quality parameters for better understanding the role of this enzyme in male fertility. MATERIALS AND METHODS: 1.5-2 yr-old healthy, 14 Merino rams were involved. Ejaculate samples were collected using an artificial vagina, in October (the breeding season) and April (the non-breeding season). Blood samples were collected prior to sperm collection. Sperm volume (ml), motility (%), mass activity (1-5), concentration (×106), viability (%), abnormal acrosome morphology (%) and abnormal sperm morphology (%) was evaluated. The activities of spermatozoa and erythrocyte G6PD were determined and the relation of sperm parameters with G6PD activity was evaluated. RESULTS: Erythrocyte G6PD activity was higher (p≤0.001), whereas spermatozoa G6PD activity was lower (p≤0.001) in the breeding season (1.928±0.231 U/g hemoglobin, 129.65±28.41 U/g protein, respectively) from that in the non-breeding (0.530±0.066 U/g hemoglobin, 562.36±94.92 U/g protein, respectively). There were also significant differences among sperm quality parameters within the seasons. Positive correlation was determined between spermatozoa G6PD activity (r=0.053, p=0.03 and sperm concentration in the breeding season. CONCLUSION: Higher spermatozoa G6PD activity in October, where the level of polyunsaturated fatty acids is suggested to be increased, may reflect the increased need of nicotinamide adenine dinucleotide phosphate and thus higher G6PD activity for the oxidative balance.

Neuroprotective effects of acetyl-l-carnitine on lipopolysaccharide-induced neuroinflammation in mice: Involvement of brain-derived neurotrophic factor.

Neuroinflammation is the inflammation of nervous tissue that can lead to neurodegeneration. Brain-derived neurotrophic factor (BDNF) is a neurotrophin which affects growth, function and survival of neurons, enhances the stabilization of synapses, regulates synaptic function and branching of dendrites and axons. Brain-derived neurotrophic factor is believed to be involved in the pathophysiology of central nervous system (CNS) diseases associated with neuroinflamation. The aim of this study was to investigate new protective and therapeutic effect of acetyl-l-carnitine (ALCAR) in neuroinflammation. Acetyl-l-carnitine was administered into Swiss Albino mice as 100mg/kg/day and 300mg/kg/day for 5days. Neuroinflammation was induced by lipopolysaccharide (LPS). Histopathological findings associated with ALCAR administration on neuroinflammation in the brain were determined. Moreover, the effects of ALCAR on BDNF concentration in the brain tissue was evaluated. The LPS administration showed higher microglial activation in the brain of LPS, 100A+LPS and 300A+LPS groups compared to that in the control (p<0.05). In the 100A+LPS group, microglial activation was lower and BDNF concentration was higher than in the 300A+LPS group (p>0.05). The findings suggest that the dose of ALCAR at 100mg/kg/day i.p. may have a beneficial effect on LPS-induced neuroinflammation in mice. As a conclusion, ALCAR may be used as an optional neuroprotective and therapeutic agent to attenuate inflammatory damage in the CNS regarding BDNF, in a dose dependent manner.