RESUMEN
In diabetes, increased oxidative stress and impaired trace element metabolism play an important role in the pathogenesis of diabetic nephropathy. The objective of this research was to examine the outcomes of blocking the renin-angiotensin system, using either the angiotensin-converting enzyme inhibitor (ACEI), perindopril, or the angiotensin II type 1 (AT1) receptor blocker, irbesartan, on oxidative stress and trace element levels such as Zn, Mg, Cu, and Fe in the kidneys of diabetic rats that had been induced with streptozotocin. Thirty-two Wistar albino male rats were equally divided into four groups. The first group was used as a control. The second group of rats developed diabetes after receiving a single intraperitoneal dose of STZ. The third and fourth groups of rats had STZ-induced diabetes and received daily dosages of irbesartan (15 mg/kg b.w/day) and perindopril (6 mg/kg b.w/day) treatment, respectively. Biochemical analysis of the kidneys showed a distinct increase in oxidative stress, indicated by heightened levels of malondialdehyde (MDA) and decreased superoxide dismutase (SOD) activities, as well as reduced glutathione (GSH) levels in the kidneys of diabetic rats. In the kidneys of diabetic rats, the mean levels of Fe and Cu were found to be significantly higher than those of the control group. Additionally, the mean levels of Zn and Mg were significantly lower in the diabetic rats compared to the control rats. Both perindopril and irbesartan decreased significantly MDA content and increased SOD activities and GSH levels in the kidneys of rats with diabetes. The Zn and Mg concentrations in the kidneys of diabetic rats treated with perindopril and irbesartan were markedly higher than in untreated STZ-diabetic rats, while the Cu and Fe concentrations were significantly lower. The urinary excretion of rats treated with perindopril and irbesartan showed a pronounced increase in Cu levels, along with a significant reduction in Zn and Mg levels. Although diabetic rats demonstrated degenerative morphological alterations in their kidneys, both therapies also improved diabetes-induced histopathological modifications in the kidneys. Finally, the present results suggest that manipulating the levels of Zn, Mg, Cu, and Fe - either through ACE inhibition or by blocking AT1 receptors - could be advantageous in reducing lipid peroxidation and increasing antioxidant concentration in the kidneys of diabetic rats.
Asunto(s)
Diabetes Mellitus Experimental , Nefropatías Diabéticas , Oligoelementos , Ratas , Animales , Inhibidores de la Enzima Convertidora de Angiotensina/farmacología , Inhibidores de la Enzima Convertidora de Angiotensina/uso terapéutico , Inhibidores de la Enzima Convertidora de Angiotensina/metabolismo , Irbesartán/metabolismo , Irbesartán/farmacología , Irbesartán/uso terapéutico , Antagonistas de Receptores de Angiotensina/metabolismo , Antagonistas de Receptores de Angiotensina/farmacología , Antagonistas de Receptores de Angiotensina/uso terapéutico , Perindopril/metabolismo , Perindopril/farmacología , Perindopril/uso terapéutico , Estreptozocina/metabolismo , Estreptozocina/farmacología , Estreptozocina/uso terapéutico , Ratas Wistar , Diabetes Mellitus Experimental/metabolismo , Oligoelementos/metabolismo , Oligoelementos/farmacología , Oligoelementos/uso terapéutico , Riñón/patología , Nefropatías Diabéticas/metabolismo , Bloqueadores del Receptor Tipo 1 de Angiotensina II/farmacología , Bloqueadores del Receptor Tipo 1 de Angiotensina II/uso terapéutico , Bloqueadores del Receptor Tipo 1 de Angiotensina II/metabolismo , Estrés Oxidativo , Superóxido Dismutasa/metabolismoRESUMEN
Normal tissue reactions are therapy limiting factor for the effectiveness of the radiotherapy in cancer patients. DNA repair and apoptosis are estimated to be critical players of adverse effects in response to radiotherapy. Our aim was to define the association of DNA repair (ERCC1 and XPC) and apoptotic (BCL2, CASP3 and NFKB1) gene expression, DNA damage levels, apoptosis changes and DNA repair gene variations with the risk of acute side effects in breast cancer patients. The study included 100 women with newly diagnosed breast cancer; an experimental case group (n=50) with acute side effects and the control group (n=50) without side effects. Gene expression was analyzed by reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR). Micronucleus (MN) and 8-hydroxy-2'-deoxyguanosine (8-OHdG) assays were performed to compare the DNA damage levels. Apoptosis was examined by TDT-mediated dUTP-biotin nick end-labeling (TUNEL) staining. ERCC1 rs3212986 and XPC rs3731055 polymorphisms were genotyped by real-time PCR technique. No significantly correlation of DNA repair and apoptosis gene expression and DNA damage levels with acute side effects in response to radiotherapy. Also, there was no association between apoptosis levels and acute effects. ERCC1 rs3212986 CC genotype showed a protective effect against radiotherapy-induced acute reactions (p<0.001; OR: 0.21; 95% CI= 0.08-0.52). Our results suggest that apoptosis and DNA damage levels are not associated with acute radiosensitivity. DNA repair may affect the risk of acute reactions. Further studies are needed to validate the current findings.
Asunto(s)
Neoplasias de la Mama/radioterapia , Carcinoma Ductal/radioterapia , Carcinoma Lobular/radioterapia , Proteínas de Unión al ADN/genética , Endonucleasas/genética , Rayos gamma/efectos adversos , Regulación Neoplásica de la Expresión Génica , Piel/efectos de la radiación , 8-Hidroxi-2'-Desoxicoguanosina , Adulto , Apoptosis/efectos de la radiación , Neoplasias de la Mama/genética , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Carcinoma Ductal/genética , Carcinoma Ductal/metabolismo , Carcinoma Ductal/patología , Carcinoma Lobular/genética , Carcinoma Lobular/metabolismo , Carcinoma Lobular/patología , Caspasa 3/genética , Caspasa 3/metabolismo , Fragmentación del ADN/efectos de la radiación , Reparación del ADN/efectos de la radiación , Proteínas de Unión al ADN/metabolismo , Desoxiguanosina/análogos & derivados , Desoxiguanosina/metabolismo , Endonucleasas/metabolismo , Femenino , Humanos , Etiquetado Corte-Fin in Situ , Pruebas de Micronúcleos , Persona de Mediana Edad , Subunidad p50 de NF-kappa B/genética , Subunidad p50 de NF-kappa B/metabolismo , Polimorfismo de Nucleótido Simple , Proteínas Proto-Oncogénicas c-bcl-2/genética , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Tolerancia a Radiación , Transducción de Señal , Piel/metabolismo , Piel/patologíaRESUMEN
The harmful use of alcohol is a worldwide problem involving all ages. This study aims to investigate chronic alcohol exposure related hepatotoxicity on the rat liver and possible hepatoprotective effects of boric acid. Rats were separated into 4 different groups: control, ethanol, ethanol+boric acid, and boric acid. We measured (i) malondialdehyde (MDA), total sialic acid (TSA), superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) levels, which are known to be the markers of alcohol damage; and also (ii) caspase-3, tumor necrosis factor-alpha (TNF-α), and the terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) as the markers of apoptosis. In the ethanol group, MDA, TSA, and TNF-α levels increased whereas SOD and CAT levels decreased compared with the control group. Ethanol+boric acid group MDA, TSA, caspase-3, and TNF-α levels decreased whereas SOD and CAT levels increased compared with the ethanol group. Using histopathological evaluation of light microscope images, immunohistochemical caspase-3 and TNF-α activity in the ethanol+boric acid group were shown to be decreased compared with that in the ethanol group. Our results revealed that ethanol is capable of triggering oxidative stress and apoptosis in the rat liver. We propose that boric acid is an effective compound in protecting the rat liver against ethanol.
Asunto(s)
Antioxidantes/uso terapéutico , Apoptosis , Ácidos Bóricos/uso terapéutico , Etanol/efectos adversos , Hepatopatías/tratamiento farmacológico , Animales , Antioxidantes/farmacología , Apoptosis/efectos de los fármacos , Ácidos Bóricos/farmacología , Caspasa 3/metabolismo , Etanol/sangre , Conducta Alimentaria , Etiquetado Corte-Fin in Situ , Hígado/efectos de los fármacos , Hígado/metabolismo , Hígado/patología , Hepatopatías/sangre , Hepatopatías/patología , Masculino , Ratas Sprague-Dawley , Coloración y Etiquetado , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
BACKGROUND: The aim of this experimental study was to investigate the effectiveness of intramuscular pentoxifylline in the prevention of postoperative fibrosis. MATERIAL/METHODS: We divided 16 adult Wistar albino rats into 2 equal groups: treatment and control. Both groups underwent L1 vertebral total laminectomy to expose the dura. The intramuscular treatment group received pentoxifylline. Four weeks later, epidural fibrosis was studied in both groups using electron microscopy, light microscopy, histology, biochemistry, and macroscopy. RESULTS: The evaluation of epidural fibrosis in the 2 groups according to macroscopic (p<0.01) assessment and light microscopy revealed that epidural scar tissue formation was lower in the treatment group compared to the control group (p<0.001) and the number of fibroblasts was also decreased significantly in the pentoxifylline-treated group (p<0.05). More immature fibers were demonstrated in the treatment group by electron microscopy in comparison with the control group. In biochemical analysis, a statistically significant decrease was detected in hydroxyproline, which indicates fibrosis and myeloperoxidase activity, and shows an inflammatory response (P<0.001). CONCLUSIONS: Systemic pentoxifylline application prevents postoperative epidural fibrosis and adhesions with various mechanisms. Our study is the first to present evidence of experimental epidural fibrosis prevention with pentoxifylline.
Asunto(s)
Espacio Epidural/patología , Laminectomía/efectos adversos , Pentoxifilina/farmacología , Animales , Espacio Epidural/efectos de los fármacos , Fibroblastos/efectos de los fármacos , Fibroblastos/patología , Fibroblastos/ultraestructura , Fibrosis , Hidroxiprolina/metabolismo , Masculino , Peroxidasa/metabolismo , Ratas WistarRESUMEN
AIM: The aim of this study was to evaluate the histopathological and biochemical impact and effectiveness of two hemostatic agents, Ankaferd blood stopper (ABS) and Microporous Polysaccharide Hemospheres (MPH), on epidural fibrosis in an experimental rat laminectomy model. MATERIAL AND METHODS: Twenty adult Wistar albino rats were divided into MPH-treated (n=6), ABS-treated (n=6) and control (n=8) groups. Laminectomy of the lumbar spine was performed in all animals and treatment groups were exposed to MPH and ABS while closure was applied in control group as per usual. Epidural fibrosis was evaluated in all groups macroscopically, histopathologically, biochemically and with electron microscopy four weeks later. RESULTS: Statistically, it was found that MPH-treated group had significantly less epidural fibrosis compared to ABS-treated and control groups. CONCLUSION: We compared two hemostatic agents for their propensity to cause adhesions in the present study. Our results show that MPH significantly reduces epidural scar formation and dural adhesion in a rat model of laminectomy while ABS increases postoperative fibrosis.
Asunto(s)
Espacio Epidural/patología , Técnicas Hemostáticas , Laminectomía/métodos , Extractos Vegetales/uso terapéutico , Animales , Cicatriz/metabolismo , Cicatriz/patología , Espacio Epidural/metabolismo , Fibrosis , Hidroxiprolina/metabolismo , Microesferas , Peroxidasa/metabolismo , Polisacáridos , Ratas , Ratas Wistar , Adherencias Tisulares/metabolismo , Adherencias Tisulares/patologíaRESUMEN
BACKGROUND: Oxytocin (OXY) is a well-known nonapeptide that functions in reproduction. It is also known as an antioxidant in several organs. However, little is about its role in the protection of tissue against ischemia/reperfusion injury in skeletal muscle. The aim of this study was to evaluate the protective and therapeutic antioxidant effect of oxytocin in skeletal muscle during ischemia/reperfusion (I/R) injury. METHODS: Rats were divided into 4 groups. Hindlimb ischemia was achieved by clamping the common femoral artery in 3 of the groups, but not a control group. OXY was injected before ischemia in the preoperative (preop) I/R + OXY group and after the onset of ischemia in the postoperative (postop) I/R + OXY group. Saline solution was injected in the I/R group. Limbs were rendered ischemic for 90 min. At the end of 90-min reperfusion period, skeletal muscle tissue samples were taken from the ischemic muscle for evaluation at light and transmission electron microscopic levels. Biochemical analysis was done for malonedialdehyde and glutathione levels. Caspase immunohistochemistry was applied for apoptosis. RESULTS: The light- and electron-microscopic scores of the OXY-treated groups were significantly lower than in the I/R group. The degree of tissue damage was ameliorated in the OXY-treated groups. The number of apoptotic cells was decreased in the OXY-treated groups compared with the I/R group. In OXY-treated groups, the malonedialdehyde level was lower than in the I/R group. Glutathione levels were found to be increased in the OXY-treated groups compared with the I/R group. CONCLUSIONS: Oxytocin has a protective effect against I/R injury in skeletal muscle and may reduce the incidence of compartment syndrome.
Asunto(s)
Antioxidantes/uso terapéutico , Caspasa 3/metabolismo , Malondialdehído/metabolismo , Músculo Esquelético/irrigación sanguínea , Oxitocina/uso terapéutico , Daño por Reperfusión/prevención & control , Animales , Glutatión/metabolismo , Miembro Posterior , Peroxidación de Lípido , Microscopía Electrónica de Transmisión , Músculo Esquelético/metabolismo , Músculo Esquelético/patología , Ratas , Ratas WistarRESUMEN
Ischemia-reperfusion (I/R) injury induces the generation of reactive oxygen species (ROS) which affect many organs. This study was designed to investigate the roles of melatonin and 1,25-dihydroxyvitamin D3 (VD3) on renal I/R injury. Thirty male Wistar albino rats were divided into five groups: group 1, control; group 2, right nephrectomy (RN) + I/R in the contralateral kidney; group 3, melatonin + RN + I/R; group 4, VD3 + RN + I/R; and group 5, melatonin + VD3 + RN + I/R. Melatonin (10 mg/kg), VD3 (0.5 µg/kg), and melatonin plus VD3 were injected intraperitoneally for 7 days before renal I/R. After 7 days, right nephrectomy was initially performed and left renal artery was clamped for 45 min. After 45-min reperfusion, the serum and kidney tissue samples were obtained for assays. Melatonin and VD3 had an ameliorative effect on biochemical parameters such as serum creatinine (SCr) and blood urea nitrogen (BUN). Renal tissue malondialdehyde (MDA), glutathione (GSH), nitric oxide (NO) levels, and superoxide dismutase (SOD) activity were determined. Renal I/R decreased the kidney tissue GSH levels and SOD activity and increased the NO levels as compared with control group. However, melatonin and VD3 and melatonin plus VD3 treatment significantly increased the tissue GSH levels and SOD activity and decreased the NO levels compared with those of I/R group. Meanwhile, MDA levels were not different between the control and I/R groups. But, MDA levels decreased in all treated groups compared to I/R and control groups. These data support that melatonin and VD3 have beneficial effects on renal injury.
Asunto(s)
Lesión Renal Aguda/prevención & control , Antioxidantes/uso terapéutico , Calcitriol/uso terapéutico , Melatonina/uso terapéutico , Daño por Reperfusión/prevención & control , Lesión Renal Aguda/metabolismo , Lesión Renal Aguda/patología , Animales , Antioxidantes/metabolismo , Evaluación Preclínica de Medicamentos , Riñón/metabolismo , Riñón/patología , Masculino , Ratas , Ratas Wistar , Daño por Reperfusión/metabolismo , Daño por Reperfusión/patologíaRESUMEN
OBJECTIVES: We aimed to investigate the effects of AR-A014418, a strong inhibitor specific to GSK-3beta, on neuronal apoptosis and neuroprotection in the traumatic SCI model. MATERIALS AND METHODS: In this study, three groups were generated from 36 Wistar rats; (1) control, (2) spinal cord trauma group created by clip compression technique after laminectomy, and (3) AR-A014418 (4mg/kg, i.p., DMSO) treatment group after laminectomy and spinal cord trauma. The TUNEL assay for apoptosis detection, immunohistochemical staining for bax and TGF-beta were applied in spinal cord tissues. For light microscopic examination, necrotic, and apoptotic cells were counted, and PMNL counting was applied to detect inflammation. Functional recovery was tested by field locomotor test in the 3rd and 7th days following surgery. RESULTS: In the trauma group, diffuse hemorrhage, cavitation, necrosis and edematous regions, degeneration in motor neurons and leukocyte infiltration were observed in gray matter. In the AR-A014418-treated groups, healthy cells were observed in more places compared to the trauma groups, however, cavitation, hemorrhagic, and edematous areas were seen in gray matter. In the AR-A014418-treatment groups, the number of apoptotic cells in the 3rd and 7th days (respectively; p<0.05, p<0.01), were significantly decreased compared to the trauma groups, as were the levels of bax (p<0.01) and TGF-beta 1 immunoreactivity. Results of the locomotor test were significantly increased in the treatment group (p<0.001) as compared to the trauma group. CONCLUSIONS: In this experimental spinal cord trauma model study neural apoptosis was significantly triggered in secondary damage developed after trauma, however, neurological healing was expedited by preventing mitochondrial apoptosis and reducing the inflammation by the potent inhibitor AR-A014418, which is GSK-3beta selective.
Asunto(s)
Antiinflamatorios/uso terapéutico , Apoptosis/efectos de los fármacos , Glucógeno Sintasa Quinasa 3/antagonistas & inhibidores , Paraplejía/prevención & control , Traumatismos de la Médula Espinal/tratamiento farmacológico , Tiazoles/uso terapéutico , Urea/análogos & derivados , Animales , Antiinflamatorios/farmacología , Evaluación Preclínica de Medicamentos , Cojera Animal/etiología , Cojera Animal/prevención & control , Laminectomía , Masculino , Mitocondrias/efectos de los fármacos , Mitocondrias/patología , Neuronas Motoras/efectos de los fármacos , Neuronas Motoras/patología , Necrosis , Degeneración Nerviosa/etiología , Degeneración Nerviosa/prevención & control , Proteínas del Tejido Nervioso/análisis , Paraplejía/etiología , Distribución Aleatoria , Ratas , Ratas Wistar , Médula Espinal/química , Médula Espinal/patología , Compresión de la Médula Espinal/tratamiento farmacológico , Compresión de la Médula Espinal/enzimología , Compresión de la Médula Espinal/patología , Traumatismos de la Médula Espinal/enzimología , Traumatismos de la Médula Espinal/patología , Tiazoles/farmacología , Factor de Crecimiento Transformador beta1/análisis , Urea/farmacología , Urea/uso terapéutico , Proteína X Asociada a bcl-2/análisisRESUMEN
This study was designed to evaluate the preventive role of melatonin (Mel) and 1,25-dihydroxyvitamin D3 (VD3) in biochemical and apoptotic events leading to tissue injury and renal dysfunction after ischemia-reperfusion (I/R). Thirty male Wistar rats were divided into five groups: sham-operated, I/R, Mel + I/R, VD3 + I/R, and Mel + VD3 + I/R. The rats were intraperitoneally administered with Mel (10 mg/kg), VD3 (0.5 µg/kg), or Mel (10 mg/kg) plus VD3 (0.5 µg/kg) each day at 1 week prior to ischemia. Right nephrectomy was initially performed and left renal I/R injury was induced by 45 min of bilateral renal ischemia followed by 45 min of reperfusion. After reperfusion, kidneys and blood were obtained for histopathologic and biochemical evaluation. Mel and VD3 had an ameliorative effect on biochemical parameters such as serum creatinine, blood urea nitrogen, alanine aminotransferase, aspartate aminotransferase, and apoptosis (caspase-3 and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling staining) in the kidneys against renal I/R injury in rats. Additionally, VD3 combined with Mel significantly reduced apoptotic and histological alterations when compared with Mel or VD3 alone. This preventive effect on renal tubular apoptosis was remarkable when Mel was combined with VD3.
Asunto(s)
Antioxidantes/uso terapéutico , Apoptosis/efectos de los fármacos , Calcitriol/uso terapéutico , Enfermedades Renales/prevención & control , Riñón/irrigación sanguínea , Melatonina/uso terapéutico , Daño por Reperfusión/tratamiento farmacológico , Animales , Antioxidantes/farmacología , Calcitriol/farmacología , Caspasa 3/análisis , Riñón/efectos de los fármacos , Riñón/patología , Túbulos Renales/efectos de los fármacos , Túbulos Renales/patología , Masculino , Melatonina/farmacología , Ratas , Ratas Wistar , Daño por Reperfusión/patologíaRESUMEN
In this study, we aimed to investigate the relationship among trace elements (Cu, Fe, Zn and Mg) on oxidative and anti-oxidative substances in liver and kidneys tissues in streptozotocin (STZ) diabetic rat model. The mean levels of Fe and Cu were found significantly higher in the liver and kidneys of the diabetic rats, in comparison to the control rats. On the other hand, the mean levels of Zn and Mg in the liver and kidneys of the diabetic rats were significantly lower than in the control rats. The liver and kidneys malonaldehyde (MDA) levels of the experimental group were found to be higher than in the control group (p < 0.001; p < 0.01, respectively) after 4 weeks of the experimental period. Superoxide dismutase (SOD) activities and glutathione (GSH) levels in the liver tissue of STZ-induced diabetic rats were found to be lower in the experimental group than in the control group (p < 0.01). SOD activity and GSH concentration in kidneys of the diabetic rats were significantly diminished with respect to the control group (p < 0.01). In conclusion, the present results indicate that the increase of Fe and Cu together with decreas of Zn and Mg concentration in liver and kidney of STZ-induced diabetic rats may be involved in disturbances of oxidative balance in both the tissues. Therefore, these findings may contribute to explain the role of impaired ion metabolism of some elements in the progression of diabetic oxidative complications.
Asunto(s)
Diabetes Mellitus Experimental/fisiopatología , Riñón/efectos de los fármacos , Hígado/efectos de los fármacos , Estrés Oxidativo , Estreptozocina/farmacología , Oligoelementos/análisis , Animales , Glucemia/metabolismo , Cobre/análisis , Hierro/análisis , Magnesio/análisis , Masculino , Metales/análisis , Ratas , Superóxido Dismutasa/metabolismo , Zinc/análisisRESUMEN
Current evidence strongly suggests that aberrant activation of the nuclear factor kappa B (NF-kB) signaling cascade is connected to carcinogenesis. The matrix metalloproteinases (MMP) which are also the key agents for tumor metastasis may be potent candidates for tumor diagnosis in clinics. In this in vitro study, we hypothesized that metformin with an effective dose can inhibit tumor cell proliferation and metastasis by modulating the expressions of MMP-2 and -9 and interfering with NF-kB signaling in primary breast cancer cells (PBCCs). 300 000 cells per ml were obtained from biopsies of breast tumors from five human donors. The cell viability and proliferation were tested. Immunocytochemistry was performed for MMP-2, MMP-9, and NF-kB, and enzyme-linked immunosorbent assay for NF-kB activity, quantitative real-time PCR for RELA/p65, IkBα, MMP-2, and MMP-9. Three different doses of metformin (5, 10, and 25â¯mM) (Met) reduced the viability and proliferation of PBCCs in a dose-dependent manner, maximum inhibition was observed at 25â¯mM Met. The expression of RELA/p65 was not affected by 25â¯mM Met. Nuclear immunoreactivity and activity of NF-kB reduced while cytoplasmic NF-kB (p65) elevated by 25â¯mM Met compared to non-treatment (Pâ¯<⯠0.05). The expression and immunoreactivity of MMP-9 but not MMP-2 were decreased by 25 mM Met treatment, compared with the non-treatment (Pâ¯<⯠0.05). Metformin may have an essential antitumor role in the invasion and metastasis pathways of PBCCs by downregulating the MMP-9 expression blocking both the activity and nuclear translocation of NF-kB.
Asunto(s)
Antineoplásicos/farmacología , Neoplasias de la Mama , Metformina/farmacología , FN-kappa B/metabolismo , Proteínas de Neoplasias/metabolismo , Transducción de Señal , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Femenino , Humanos , Persona de Mediana Edad , Invasividad Neoplásica , Metástasis de la Neoplasia , Células Tumorales CultivadasRESUMEN
In this study, we investigated the protective effect of losartan as an AT1 receptor antagonist by evaluating the expression of apoptosis-regulatory genes that contribute to the progressive damage in the renal tubules of hyperoxaluric rats. Rats were divided into 4 groups of 10 each; control (C), ethylene glycol (EG), ethylene glycol + losartan (EG + L) and Losartan (L). For 4 weeks 0.8% EG, as a precursor for oxalate, was administered to EG and EG + L and losartan (300 mg/l) was administered to groups EG + L and L. Urine and blood samples were collected for biochemical determination. Bcl-2, bax, caspase-3 and TGF-beta 1 antibodies were used for immunohistochemistry. Apoptosis was determined by TUNEL method. A marked increase in urinary oxalate levels of the rats in EG and EG + L groups was found. In the EG group a diffuse amount of oxalate crystals into the tubular lumina and interstitium in the cortex was observed. In the EG group GBM thickening, interstitial fibrosis and tubular atrophy with infiltration of mononuclear cell findings reduced in the EG + L group were presented as well. In the EG group, immunoreactivity of TGF-beta 1 was increased in glomeruli and tubuli. In the EG + L group, immunoreactivity of TGF-beta 1 was decreased compared to the EG group. Bax expression increased in the renal tubules of EG group and reduced in the EG + L group comparing to the control. In the EG + L group, the immunoreactivity of bcl-2 was increased in glomeruli. In EG + L treated group, number of caspase-3 immunopositive cells were decreased compared to all groups (P < 0.01). Apoptotic cells were increased in the EG-treated group compared to the other groups. Decreased apoptotic cell number was observed in the EG + L compared to the EG group (P < 0.01). Our findings suggest that losartan may provide a beneficial effect against tubulointerstitial damage and decrease renal tubular apoptosis caused by hyperoxaluria.
Asunto(s)
Bloqueadores del Receptor Tipo 1 de Angiotensina II/farmacología , Apoptosis/efectos de los fármacos , Apoptosis/genética , Hiperoxaluria/genética , Túbulos Renales/citología , Túbulos Renales/efectos de los fármacos , Losartán/farmacología , Animales , Masculino , Ratas , Ratas WistarRESUMEN
Several epidemiological studies have shown that exposure to electromagnetic radiation (EMR) can be harmful to human health. The purpose of this study was to examine oxidative parameters and apoptosis induced by EMR in human kidney embryonic cells (HEK293) and to investigate whether zinc (Zn) has protective effect on EMR-induced apoptosis in HEK293 cells. For our experiment, HEK293 cells were divided into four main groups, control, EMR, 50 µM Zn + EMR, and 100 µM Zn + EMR. HEK293 cells of EMR groups were exposed to 2.45 GHz EMR for 1 h. In Zn groups, HEK293 cells were incubated with different concentrations of Zn for 48 h before EMR exposure. Oxidative stress parameters were determined by spectrophotometric method; bcl-2 and caspase-3 were assessed immunohistochemically and TUNEL method was performed for apoptotic activity. EMR group had higher malondialdehyde (MDA) level and lower superoxide dismutase (SOD) activity compared with control group. In Zn-applied groups, MDA was decreased and SOD activity was increased compared with EMR group. The number of the apoptotic cells and caspase-3 immunopositive cells at EMR group was increased significantly compared with the control group, whereas bcl-2 was decreased. Besides, Zn-treated groups showed a significant reduction in the number of apoptotic cells and caspase-3 from that of EMR group, whereas there was an increase in bcl-2 immunopositivity. Our findings show that EMR caused oxidative stress and apoptotic activation in HEK293 cells. Zn seems to have protective effects on the EMR by increasing SOD activity and bcl-2 immunopositivity, decreasing lipid peroxidation and caspas-3 immunopositivity.
Asunto(s)
Teléfono Celular , Zinc , Antioxidantes , Apoptosis , Radiación Electromagnética , Células HEK293 , Humanos , Malondialdehído , Estrés Oxidativo , Zinc/farmacologíaRESUMEN
Bisphenol A (BPA) is a key endocrine-disrupting chemical (EDC) in the manufacturing industry. It is found in the structure of compounds such as polycarbonate and epoxy in combination with other chemicals. Our objective was to investigate the effect of BPA on rat ovaries. A total of 32 female rats were divided into four equal groups: In group 1 (control), vehicle was administered; in group 2, BPA 50 µg/day was administered intraperitoneally; in group 3, BPA 100 mg/kg/day was administered intraperitoneally; and in group 4, BPA 100 mg/kg/day and vitamin C (50 mg/kg) were administered intraperitoneally, while vitamin E (50 mg/kg) was administered intramuscularly. Thirty days after the treatment, the effects of BPA on the ovaries were evaluated by terminal deoxynucleotidyltransferase [TdT]-mediated dUTP-biotin nick end labeling (TUNEL) assay. There was no difference in the number of apoptotic cells between group 2 and group 4. In addition, there was no significant difference between control group and group 2, 4. However, the number of apoptotic cells per unit area was significantly increased in group 3 compared with all groups (p < 0.01, p < 0.05). In conclusion, this study showed that high doses of BPA (100 mg/kg/day) have a toxic effect on the ovaries. The fact that the number of apoptotic cells in the group administered with high dose of BPA + 50 mg/kg/day vitamin C + 50 mg/kg/day vitamin E was lower than that of the high-dose BPA-administered group shows that these vitamins may have a protective effect.
Asunto(s)
Compuestos de Bencidrilo/toxicidad , Disruptores Endocrinos/toxicidad , Ovario/efectos de los fármacos , Fenoles/toxicidad , Animales , Apoptosis , Ácido Ascórbico/farmacología , Femenino , Etiquetado Corte-Fin in Situ , Masculino , Ratas , Pruebas de Toxicidad , Vitamina E/farmacologíaRESUMEN
BACKGROUND: Apoptosis plays a major role in fatty liver disease. High-fat diets are related to the onset of fatty liver disease and hepatic oxidant-antioxidant imbalance. Curcumin and capsaicin are somewhat beneficial in reducing hepatic triglycerides; this is most likely because they are known to downregulate reactive oxygen species (ROS) and apoptosis. OBJECTIVES: The aim of this study was to investigate the effects of curcumin and capsaicin on apoptosis through the oxidative effect in an animal model of fatty liver disease. MATERIAL AND METHODS: Male Sprague Dawley rats were fed a normal control diet, a high-fat diet (HFD; 60% of total calories from fat), a HFD+curcumin (1.5 g curcumin/kg HFD), a HFD+capsaicin (0.15 g capsaicin/kg HFD), or a HFD+curcumin+capsaicin (1.5 g curcumin and 0.15 g capsaicin/kg HFD). Liver lysate levels of BAX, Bcl-2 and caspase-3 were determined via immunoblotting. Caspase-3 activity was measured with a colorimetric caspase-3 measurement kit. Total antioxidant status (TAS) and total oxidant status (TOS) were assayed using commercial kits. The generation of hepatic ROS was measured with fluorimetry. Fragmentation of DNA was detected using the TUNEL method. RESULTS: High-fat diet caused increased expression of BAX and caspase-3, as well as increased TOS and caspase-3 activity, but decreased expression of Bcl-2. HFD+curcumin+capsaicin caused decreased BAX, caspase-3, TOS, and ROS levels as compared to HFD, but increased TAS and Bcl-2. A HFD +curcumin+capsaicin also decreased the number of TUNEL-positive cells. CONCLUSIONS: These results suggest that supplementation with curcumin and capsaicin balances the hepatic oxidant-antioxidant status and may have a protective role in the apoptotic process in an HFD-induced fatty liver model.
Asunto(s)
Antiinflamatorios no Esteroideos , Capsaicina , Curcumina , Dieta Alta en Grasa , Estrés Oxidativo , Animales , Antiinflamatorios no Esteroideos/farmacología , Apoptosis , Capsaicina/farmacología , Curcumina/farmacología , Hígado , Masculino , Ratas , Ratas Sprague-DawleyRESUMEN
Quercetin (QR) is part of a subclass of flavonoids called flavonols. We aimed to investigate the effect of QR on malondialdehyde (MDA), advanced oxidation protein products (AOPPs), and glutathione peroxidase (GSH-Px) activity in the liver of diabetic rats. We compared four groups of male adult Wistar albino rats: a control group, an untreated diabetic group, diabetic groups treated with QR, and QR group. Diabetes was induced by a single injection of streptozotocin (STZ) (50 mg/kg). Animals were kept in standard condition. On the 31st day of the study, the liver tissue was removed for biochemical parameters and histopathological evaluations. In an untreated diabetic group, liver MDA and AOPP levels were significantly higher than all groups. QR treatment significantly decreased the increased MDA, AOPP levels, and increased the decreased GSH-Px enzyme activity in liver tissues. The QR-treated rats in the diabetic group showed an improved histological appearance. Lesser vesicular vacuolization and fibrotic areas were observed in the QR-treated diabetic group than in the diabetic group. The STZ-induced liver injury is associated with oxidative stress, and coadministration of QR may reduce this damage effectively in a rat model. Our results are also supported by morphological improvement in liver tissue. Therefore, we think QR may be effective in treating hyperglycemia and oxidative damage in diabetes.
Asunto(s)
Antioxidantes/farmacología , Diabetes Mellitus Experimental/metabolismo , Peroxidación de Lípido/efectos de los fármacos , Hígado/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Carbonilación Proteica/efectos de los fármacos , Quercetina/farmacología , Animales , Antioxidantes/metabolismo , Antioxidantes/uso terapéutico , Diabetes Mellitus Experimental/tratamiento farmacológico , Diabetes Mellitus Experimental/patología , Hígado/metabolismo , Masculino , Malondialdehído , Extractos Vegetales/farmacología , Extractos Vegetales/uso terapéutico , Quercetina/uso terapéutico , Ratas WistarRESUMEN
AIMS: Lucilia sericata larvae have been successfully used on healing of wounds in the diabetics. However, the involvement of the extraction/secretion (ES) products of larvae in the treatment of diabetic wounds is still unknown. Activator protein-1 (AP-1) transcription, composed of c-jun and c-Fos proteins, has been shown to be the principal regulator of multiple MMP transcriptions under a variety of conditions, also in diabetic wounds. Specifically, MMP-2 and MMP-9's transcriptions are known to be modulated by AP-1. c-jun has been demonstrated to be a repressor of p53 in immortalized fibroblasts. The aim of the present study is to investigate the effects of L. sericata ES on the expression of AP-1 (c-jun), p53, MMP-2, and MMP-9 in wound biopsies dissected from streptozotocin induced diabetic rats. METHODS: The expression levels of MMP-2, MMP-9, c-jun and p53 in dermal tissues were determined at days 0, 3, 7 and 14 after wounding, using immunohistochemical analysis and quantitative real-time PCR. RESULTS: The treatment with ES significantly decreased through inflammation-based induction of MMP-2 and MMP-9 expression levels in the wounds of diabetic groups, compared to control groups at the third day of wound healing. At the 14th day, there were dramatic decreases in expression of c-jun, MMP-9, and p53 in ES-treated groups, compared to the diabetic group (P < 0.001, P < 0.05 and P < 0.01, respectively). CONCLUSION: ES products of L. sericata may enhance the process of wound healing in phases of inflammation, proliferation, and re-epithelization, essentially via regulating c-jun expression and modulating MMP-2 and MMP-9 expressions.
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Diabetes Mellitus Experimental , Pie Diabético , Dípteros , Proteínas de Insectos/farmacología , Metaloproteinasa 2 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Factor de Transcripción AP-1/metabolismo , Animales , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Experimental/terapia , Pie Diabético/metabolismo , Pie Diabético/terapia , Fibroblastos/metabolismo , Fibroblastos/patología , Larva , Masculino , Ratas , Transducción de Señal/fisiología , Resultado del Tratamiento , Cicatrización de Heridas/efectos de los fármacos , Cicatrización de Heridas/fisiologíaRESUMEN
Our first aim was to determine the effects of secreted clusterin (sCLU) and nuclear clusterin (nCLU) in diabetic nephropathy. We also aimed to investigate the post-effects of angiotensin II blockage treatment on clusterin expression and to compare these with apoptosis. Five groups of Wistar albino rats were used: First group consisted of healthy controls; the second group included the untreated STZ-diabetics; 30 days of irbesartan or perindopril treated STZ-diabetics formed the third and the fourth groups, respectively; while the subjects receiving a combined treatment with irbesartan and perindopril for 30 days consisted the fifth group. TUNEL method for apoptosis and immunohistochemical staining for TGF-beta1, alpha-SMA, clusterin-beta and clusterin-alpha/beta antibodies were performed. Apoptotic cells especially increased in the kidney tubuli of untreated diabetic group and on the contrary, a significant decrease was observed in the group that received a combined drug treatment. While sCLU was increased in the glomeruli and tubuli of the untreated diabetic group, it was decreased in all the treated groups. An increase in the nCLU immunoreactivity was observed in the podocytes, mesangial cells, and the injured tubule cells of the untreated diabetic group. nCLU immunopositive cells were decreased in all treated diabetic groups. In addition to this, the distribution of nCLU was similar to the distribution of apoptotic cells in the diabetic groups. Our results indicate that sCLU expression in diabetic nephropathy was induced due to renal tissue damage, and the nCLU expression increase in renal tubuli was related to apoptosis. Although irbesartan and perindopril prevented further renal injury in diabetes, a combined application of low-dose ACEI and AT1R blockers revealed more efficient measures, by means of renal damage prevention.
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Bloqueadores del Receptor Tipo 1 de Angiotensina II/farmacología , Inhibidores de la Enzima Convertidora de Angiotensina/farmacología , Apoptosis/fisiología , Clusterina/metabolismo , Riñón/efectos de los fármacos , Isoformas de Proteínas/metabolismo , Actinas/metabolismo , Animales , Glucemia/metabolismo , Peso Corporal , Diabetes Mellitus Experimental , Etiquetado Corte-Fin in Situ , Riñón/citología , Riñón/fisiología , Masculino , Ratas , Ratas Wistar , Factor de Crecimiento Transformador beta/metabolismoRESUMEN
There is a widespread use of 2.4â¯GHz electromagnetic radiation emitting devices especially in communication and education. Recent studies show the adverse effects of electromagnetic fields (EMF) such as oxidative stress, cellular damage and apoptosis on tissues. Selenium (Se) has an antioxidant properties by inhibiting oxidative damage being within the structure of antioxidant enzymes like glutathione peroxidase (GSH-Px) and it has also regulatory function for cell cycle and apoptosis. The aim of this study was to investigate the effect of Se on 2.4â¯GHz frequency EMF exposed human embryonic kidney cells (HEK293) by means of alterations in apoptotic and oxidative stress parameters. Our study was planned as control, EMF, 100â¯nM Seâ¯+â¯EMF, 200â¯nM Seâ¯+â¯EMF groups. EMF groups were exposed to 2.4â¯GHz EMF for 1â¯h, element groups were incubated with two different doses of Se added cell culture medium for 48â¯h before EMF exposure. MDA levels were significantly higher whereas SOD and GSH-Px activities were significantly lower in EMF compared to control. 100 and 200â¯nM Seâ¯+â¯EMF application decreased MDA levels, increased SOD and GSH-Px activities than EMF. Apoptosis and caspase-3 were statistically significantly higher but bcl-2 was lower in EMF than control. Apoptosis and caspase-3 were lower in 100 and 200â¯nM Seâ¯+â¯EMF, although bcl-2 were higher than EMF. In conclusion, Se has protective effects against 2.4â¯GHz EMF-induced oxidative stress by reducing lipid peroxidation, regulating SOD and GSH-Px activity. Also, Se has inhibitory effect on 2.4â¯GHz EMF induced apoptosis by increasing the expression of anti-apoptotic protein bcl-2 and suppressing apoptosis regulatory protein caspase-3.
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Campos Electromagnéticos/efectos adversos , Estrés Oxidativo/efectos de los fármacos , Estrés Oxidativo/efectos de la radiación , Selenio/farmacología , Antioxidantes/metabolismo , Apoptosis/efectos de los fármacos , Apoptosis/efectos de la radiación , Glutatión Peroxidasa/metabolismo , Células HEK293 , Humanos , Peroxidación de Lípido/efectos de los fármacos , Peroxidación de Lípido/efectos de la radiación , Malondialdehído/metabolismo , Oxidación-Reducción/efectos de los fármacos , Oxidación-Reducción/efectos de la radiaciónRESUMEN
Metformin is the most widely used anti-diabetic drug in the world. It reduces advanced glycation end product (AGEs)-induced ROS generation in high glucose condition. Protein glycation contributes to skin aging as it deteriorates the existing collagen by crosslinking. The progressive increase of AGE during aging not only causes oxidative damage to cellular macromolecules but also modulates the activation of transcription factors nuclear factor kappa-B(NF-kB). However, it is still unclear whether metformin can change collagen production and NF-kB activity induced by high glucose conditions in 3T3 fibroblast. The effects of metformin on proliferation, apoptosis, and collagen levels and NF-kB activity of in vitro cell aging model of 3T3 fibroblast cells in high glucose conditions. At first, we investigated the effects of 50 mM high glucose concentration, with or without metformin, on 3T3 fibroblast proliferation, by BrdU immunostaining for cell proliferation. Apoptotic levels were analyzed by flow cytometric assay. NF-kB(p65) activity was measured by transcription factor assay kit and collagen I and III levels by Collagen Estimation Assay through ELISA. We observed that metformin exposure leads to decreased apoptosis levels and increased proliferation of 3T3 fibroblast in high glucose media. We also determined that metformin exposure leads to increased production of collagen I-III and decreased activation of NF-kB(p65) activity. The data are consistent with the observation that metformin has a protective effect in this in vitro model of aging 3T3 fibroblasts under high glucose conditions inducing cell proliferation, collagen I and III production, protection from apoptosis, and reducing NF-kB(p65) activity.