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For economic purposes, cooking oil is repeatedly heated in food preparation, which imposes serious health threats. This study investigated the detrimental effects of multiple-heated cooking oil (MHO) on hepatic and renal tissues with particular focusing on cellular senescence (CS), and the potential regenerative capacity of oleuropein (OLE). Adult male rats were fed MHO-enriched diet for 8 weeks and OLE (50 mg/kg, PO) was administered daily for the last four weeks. Liver and kidney functions and oxidative stress markers were measured. Cell cycle markers p53, p21, cyclin D, and proliferating cell nuclear antigen (PCNA) were evaluated in hepatic and renal tissues. Tumor necrosis factor-α (TNF-α) and Bax were assessed by immunohistochemistry. General histology and collagen deposition were also examined. MHO disturbed hepatic and renal structures and functions. MHO-fed rats showed increased oxidative stress, TNF-α, Bax, and fibrosis in liver and kidney tissues. MHO also enhanced the renal and hepatic expression of p53, p21, cyclin D and PCNA. On the contrary, OLE mitigated MHO-induced oxidative stress, inflammatory burden, apoptotic and fibrotic changes. OLE also suppressed CS and preserved kidney and liver functions. Collectively, OLE displays marked regenerative capacity against MHO-induced hepatic and renal CS, via its potent antioxidant and anti-inflammatory effects.
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High fat diet (HFD) consumption can cause dysregulation of glucose and lipid metabolism, coupled with increased ectopic lipid deposition in renal tissue leading to steatosis and dysfunction. Sitagliptin is a dipeptidyl peptidase-4 (DPP-4) inhibitor clinically used for type II diabetes therapy; however its effect on renal steatosis in obese state is still uncertain. Herein, obesity was induced by feeding male Wistar rats HFD for 18 weeks, thereafter received either drug vehicle, or sitagliptin (10 mg/kg, PO) along with HFD for further 6 weeks and compared with age-matched rats receiving normal chow diet (NCD). After 24 weeks, serum and kidneys were collected for histological and biochemical assessments. Compared to NCD-fed group, HFD-fed rats displayed marked weight gain, increased fat mass, insulin resistance, dyslipidemia, impaired kidney functions and renal histological alterations. Sitagliptin effectively ameliorated obesity and related metabolic perturbations and improved kidney architecture and function. There were increased levels of triglycerides and cluster of differentiation 36 (CD36) in kidneys of obese rats, that were lowered by sitagliptin therapy. Sitagliptin significantly repressed the expression of lipogenesis genes, while up-regulated genes involved in mitochondrial biogenesis and fatty acid oxidation in kidneys of HFD-fed rats. Sitagliptin was found to induce down-regulation of endoplasmic reticulum (ER) stress and apoptotic markers in kidneys of obese rats. These findings together may emphasize a novel concept that sitagliptin can be an effective therapeutic approach for halting obesity-related renal steatosis and CKD.
Assuntos
Antígenos CD36 , Dieta Hiperlipídica , Estresse do Retículo Endoplasmático , Rim , Obesidade , Transdução de Sinais , Fosfato de Sitagliptina , Animais , Masculino , Ratos , Antígenos CD36/metabolismo , Antígenos CD36/genética , Dieta Hiperlipídica/efeitos adversos , Inibidores da Dipeptidil Peptidase IV/farmacologia , Inibidores da Dipeptidil Peptidase IV/uso terapêutico , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Rim/efeitos dos fármacos , Rim/patologia , Rim/metabolismo , Nefropatias/tratamento farmacológico , Nefropatias/metabolismo , Nefropatias/etiologia , Nefropatias/patologia , Nefropatias/prevenção & controle , Obesidade/tratamento farmacológico , Obesidade/metabolismo , Obesidade/complicações , Ratos Wistar , Transdução de Sinais/efeitos dos fármacos , Fosfato de Sitagliptina/farmacologia , Fosfato de Sitagliptina/uso terapêuticoRESUMO
Sunitinib (SUN) is a chemotherapeutic agent clinically approved for treatment of metastatic renal carcinoma. Despite its remarkable benefits, various renal toxicities have been reported that limit its clinical uses. Oleuropein (OLE) is the main polyphenolic constituent of olive tree and mediates the majority of its valuable pharmacological activities. The current study examined the probable renoprotective effects of OLE against SUN-induced nephrotoxicity. Adult male albino rats were co-treated by SUN (25 mg/kg, 3 times/week, PO) with either a drug vehicle or OLE (60 mg/kg/day, daily, PO) for four weeks. A control group comprising of age-matched rats was used. Four weeks later, blood specimens were collected to assess kidney functions. Kidneys were harvested for biochemical and histopathological analyses. Administration of SUN induced kidney dysfunction, along with marked rises in endothelin-1 (ET-1) and monocyte chemotactic protein-1 (MCP-1) levels in renal tissues. Histological abnormalities were also detected in kidneys of SUN-treated rats including glomerular and tubular interstitial congestion along with interstitial fibrosis. On molecular levels, there was a decline in renal SIRT6 expression along with significant up-regulation of Notch-1, NLRP-3, interleukin -1ß (IL-1ß) and cleaved caspsase-3. All these changes were almost alleviated by OLE co-treatment. These findings suggest the implication of SIRT6/Notch-1/NLRP3/IL-1ß axis in the pathogenesis of SUN-induced nephrotoxicity and highlight OLE as a prospective renoprotective agent during SUN chemotherapy to halt its renal toxicity likely through promotion of SIRT6 and suppression of Notch-1/NLRP3/IL-1ß signaling pathway.
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Metabolic syndrome (MetS) is largely coupled with chronic kidney disease (CKD). Glycogen synthase kinase-3ß (GSK-3ß) pathway drives tubular injury in animal models of acute kidney injury; but its contribution in CKD is still elusive. This study investigated the effect empagliflozin and/or pirfenidone against MetS-induced kidney dysfunction, and to clarify additional underpinning mechanisms particularly the GSK-3ß signaling pathway. Adult male rats received 10%w/v fructose in drinking water for 20 weeks to develop MetS, then treated with either drug vehicle, empagliflozin (30 mg/kg/day) and/or pirfenidone (100 mg/kg/day) via oral gavage for subsequent 4 weeks, concurrently with the high dietary fructose. Age-matched rats receiving normal drinking water were used as controls. After 24 weeks, blood and kidneys were harvested for subsequent analyses. Rats with MetS showed signs of kidney dysfunction, structural changes and interstitial fibrosis. Activation of GSK-3ß, decreased cyclinD1 expression and enhanced apoptotic signaling were found in kidneys of MetS rats. There was abundant alpha-smooth muscle actin (α-SMA) expression along with up-regulation of TGF-ß1/Smad3 in kidneys of MetS rats. These derangements were almost alleviated by empagliflozin or pirfenidone, with evidence that the combined therapy was more effective than either individual drug. This study emphasizes a novel mechanism underpinning the beneficial effects of empagliflozin and pirfenidone on kidney dysfunction associated with MetS through targeting GSK-3ß signaling which can mediate the regenerative capacity, anti-apoptotic effects and anti-fibrotic properties of such drugs. These findings recommend the possibility of using empagliflozin and pirfenidone as promising therapies for management of CKD in patients with MetS.
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Compostos Benzidrílicos , Glucosídeos , Glicogênio Sintase Quinase 3 beta , Túbulos Renais , Síndrome Metabólica , Piridonas , Animais , Piridonas/farmacologia , Masculino , Glucosídeos/farmacologia , Glucosídeos/uso terapêutico , Compostos Benzidrílicos/farmacologia , Glicogênio Sintase Quinase 3 beta/metabolismo , Ratos , Síndrome Metabólica/tratamento farmacológico , Síndrome Metabólica/complicações , Túbulos Renais/efeitos dos fármacos , Túbulos Renais/patologia , Túbulos Renais/metabolismo , Regeneração/efeitos dos fármacos , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacosRESUMO
Objectives: Distant liver injury is a complication of renal ischemia-reperfusion (I/R) injury, which imposes mortality and economic burden. This study aimed to elucidate the cross-talk of endoplasmic reticulum (ER) stress and mitochondrial perturbations in renal I/R-induced liver injury, and the potential hepatoprotective effect of azilsartan (AZL).Methods: Male albino Wister rats were pre-treated with AZL (3 mg/kg/day, PO) for 7 days then a bilateral renal I/R or sham procedure was performed. Activities of liver enzymes were assessed in plasma. The structure and ultra-structure of hepatocytes were assessed by light and electron microscopy. Markers of ER stress, mitochondrial biogenesis and apoptosis were analyzed in livers of rats.Results: Renal ischemic rats showed higher plasma levels of liver enzymes than sham-operated rats, coupled with histological and ultra-structural alterations in hepatocytes. Mechanistically, there was up-regulation of ER stress markers and suppression of mitochondrial biogenesis-related proteins and enhanced apoptosis in livers of renal ischemic rats. These abnormalities were almost abrogated by AZL pretreatment.Discussion: Our findings uncovered the involvement of mitochondrial perturbations, ER stress and apoptosis in liver injury following renal I/R, and suggested AZL as a preconditioning strategy to ameliorate remote liver injury in patients susceptible to renal I/R after adequate clinical testing.
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Benzimidazóis , Nefropatias , Oxidiazóis , Traumatismo por Reperfusão , Humanos , Ratos , Masculino , Animais , Isquemia , Traumatismo por Reperfusão/tratamento farmacológico , Traumatismo por Reperfusão/patologia , Fígado/metabolismo , Reperfusão , Apoptose , Estresse do Retículo EndoplasmáticoRESUMO
This study investigated the therapeutic effect of linagliptin and/or vitamin D3 on testicular steroidogenesis and spermatogenesis in cisplatin-exposed rats including their impact on endoplasmic reticulum (ER) stress and NF-κB/iNOS crosstalk. Cisplatin (7 mg/kg, IP) was injected into adult male albino rats which then were orally treated with drug vehicle, linagliptin (3 mg/kg/day), vitamin D3 (10 µg/kg/day) or both drugs for four weeks. Age-matched rats were used as the control group. Serum samples and testes were collected for further analyses. Cisplatin induced testicular weight loss, deteriorated testicular architecture, loss of germ cells and declined serum and intra-testicular testosterone levels, compared to the control group. There was down-regulation of steroidogenic markers including StAR, CYP11A1, HSD3b and HSD17b in cisplatin-exposed rats, compared with controls. Cisplatin-exposed rats showed up-regulation of ER stress markers in testicular tissue along with increased expression of NF-κB and iNOS in spermatogenic and Leydig cells. These perturbations were almost reversed by vitamin D3 or linagliptin. The combined therapy exerted a more remarkable effect on testicular dysfunction than either monotherapy. These findings suggest a novel therapeutic application for linagliptin combined with vitamin D3 to restore testicular architecture, aberrant steroidogenesis and spermatogenesis after cisplatin exposure. These effects may be attributed to suppression of ER stress and NF-kB/iNOS.
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Cisplatino , Testículo , Animais , Masculino , Colecalciferol/farmacologia , Cisplatino/farmacologia , Estresse do Retículo Endoplasmático , Linagliptina , NF-kappa B/metabolismo , Estresse Oxidativo , Espermatogênese , RatosRESUMO
AIMS: High dietary fructose consumption has been linked to the development of renal fibrosis. Dulaglutide is a long acting glucagon like peptide-1 (GLP-1) analog, showing some renoprotective properties; however its action on renal fibrosis remains uncertain. We investigated the effect of dulaglutide on fructose-induced renal fibrosis in comparison to pirfenidone, as well-established anti-fibrotic drug, and the contribution of epithelial-mesenchymal transition (EMT) process and its upstream signaling. MAIN METHODS: Six week-old male Wistar albino rats received 10%w/v fructose solution in drinking water for 24 weeks and co-treated with either pirfenidone (100 mg/kg/day, orally) or dulaglutide (0.2 mg/kg/week, s.c) for the last four weeks. Lipid profile, glucose homeostasis, kidney functions were assessed. Kidneys were harvested for biochemical and histological analyses. KEY FINDINGS: High dietary fructose consumption for 24 weeks induced insulin resistance, dyslipidemia and renal dysfunction that were ameliorated by dulaglutide and pirfenidone to lesser extent. Histological examination revealed histological lesions and interstitial fibrosis in renal sections of high fructose-fed rats, which were reversed by dulaglutide or pirfenidone treatment. Both drugs modulated the EMT-related proteins by increasing the epithelial marker, E-cadherin, while suppressing the mesenchymal markers, vimentin and alpha-smooth muscle actin (α-SMA) in renal tissue. Moreover, both drugs attenuated fructose-induced upregulation of GSK-3ß/TGF-ß1/Smad3 signaling. SIGNIFICANCE: These findings suggest that dulaglutide can emerge as a promising therapeutic agent for fructose-induced renal fibrosis. These results add mechanistic insights into the anti-fibrotic action of dulaglutide through suppressing EMT and the upstream GSK-3ß/TGF-ß1/Smad3 signaling.
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Água Potável , Nefropatias , Animais , Ratos , Masculino , Fator de Crescimento Transformador beta1/metabolismo , Transição Epitelial-Mesenquimal , Glicogênio Sintase Quinase 3 beta/metabolismo , Vimentina/metabolismo , Actinas/metabolismo , Frutose/farmacologia , Ratos Wistar , Fibrose , Nefropatias/tratamento farmacológico , Transdução de Sinais , Caderinas/metabolismo , Peptídeo 1 Semelhante ao Glucagon/farmacologia , Glucose/farmacologia , Lipídeos/farmacologiaRESUMO
Aging and major chronic diseases are risk factors for lower urinary tract symptoms (LUTS). On the other hand, oxidative stress (OS) is one of the fundamental mechanisms of aging and the development of chronic diseases. Therefore, OS might be a candidate mechanism linking these two clinical entities. This article aims to summarize the studies on the prevalence of LUTS, the role of OS in aging and chronic diseases, and the potential mechanisms supporting the putative link. A comprehensive literature search was performed to identify recent reports investigating LUTS and OS in major chronic diseases. In addition, studies on the impact of OS on the lower urinary tract, including bladder, urethra, and prostate, were collected and summarized. Many studies showed LUTS are prevalent in aging and major chronic diseases, including obesity, metabolic syndrome, diabetes, cardiovascular disease, hypertension, obstructive sleep apnea, autoimmune diseases, Alzheimer's disease, and Parkinson's disease. At the same time, OS is a key component in the pathogenesis of those chronic diseases and conditions. Recent studies also provided evidence that exacerbated OS can cause functional and/or structural changes in the bladder, urethra, and prostate, leading to LUTS. The reviewed data support the concept that OS is involved in multiple risk factors-associated LUTS, although further studies are needed to confirm the causative relationship. The specific ROS/RNS and corresponding reactions/pathways involved in chronic diseases and associated LUTS should be identified in the future and could serve as therapeutic targets.
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Decreased activity of AMP-activated protein kinase (AMPK) is implicated in the pathogenesis of diabetic cardiomyopathy (DCM). Recent evidence suggests a crosstalk between cinacalcet and AMPK activation. This study investigated the effects of cinacalcet on cardiac remodeling and dysfunction in type 2 diabetic rats (T2DM). High fat diet for 4 weeks combined with single intraperitoneal injection of streptozotocin (30 mg/kg) was used to induce type 2 diabetes in rats. Diabetic rats were either orally treated with vehicle, 5 or 10 mg/kg cinacalcet for 4 weeks. Control rats were fed standard chow diet and intraperitoneally injected with citrate buffer. T2DM rats showed lower body weight (BW), hyperglycemia and dyslipidemia, along with increased heart weight (HW) and HW/BW ratio. Masson's trichrome stained cardiac sections revealed massive fibrosis in T2DM rats. There were increased TGF-ß1 and hydroxyproline levels, coupled with up-regulation of atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) in hearts of T2DM rats. These alterations were associated with redox imbalance and impaired cardiac functions. Decreased phosphorylation of AMPK at threonine172 residue was found in T2DM hearts. Cinacalcet for 4 weeks significantly activated AMPK and alleviated cardiac remodeling and dysfunction in a dose-dependent manner, without affecting blood glucose, serum calcium and phosphorus levels. Cinacalcet increased the mitochondrial DNA content, and expressions of PGC-1α, UCP-3, beclin-1 and LC3-II/LC3-I ratio. Cinacalcet decreased the pro-apoptotic Bax, while increased the anti-apoptotic Bcl-2 in cardiac tissue of T2DM rats. These findings might highlight cinacalcet as an alternative therapy to combat the development and progression of DCM.
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Proteínas Quinases Ativadas por AMP/metabolismo , Autofagia/efeitos dos fármacos , Cinacalcete/farmacologia , Diabetes Mellitus Experimental/tratamento farmacológico , Diabetes Mellitus Tipo 2/tratamento farmacológico , Cardiomiopatias Diabéticas/prevenção & controle , Mitocôndrias Cardíacas/efeitos dos fármacos , Miócitos Cardíacos/efeitos dos fármacos , Remodelação Ventricular/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Diabetes Mellitus Experimental/induzido quimicamente , Diabetes Mellitus Experimental/enzimologia , Diabetes Mellitus Experimental/fisiopatologia , Diabetes Mellitus Tipo 2/induzido quimicamente , Diabetes Mellitus Tipo 2/enzimologia , Diabetes Mellitus Tipo 2/fisiopatologia , Cardiomiopatias Diabéticas/enzimologia , Cardiomiopatias Diabéticas/etiologia , Cardiomiopatias Diabéticas/fisiopatologia , Fibrose , Hemodinâmica/efeitos dos fármacos , Masculino , Mitocôndrias Cardíacas/enzimologia , Mitocôndrias Cardíacas/patologia , Miócitos Cardíacos/enzimologia , Miócitos Cardíacos/patologia , Estresse Oxidativo/efeitos dos fármacos , Ratos Wistar , Transdução de Sinais , EstreptozocinaRESUMO
AIMS: Hemorrhagic cystitis (HC) is a major urotoxic complication of cyclophosphamide (CPA) therapy. This study investigated the uroprotective effect of montelukast on CPA-induced HC, compared to the efficacy of 2-mercaptoethane sulfonate sodium (MESNA). MAIN METHODS: Male albino rats were pretreated with MESNA (40 mg/kg/day, IP) or montelukast (10 mg/kg/day, orally) for three days then received a single dose of CPA (200 mg/kg, IP), 1 h after the last dose, and compared to CPA-treated rats receiving drug vehicle. Age-matched rats were used as controls. Bladders of rats were assessed biochemically, macroscopically and microscopically by light and electron microscope 24 h later. KEY FINDINGS: CPA injection contributed to increased bladder weight, urothelial ulceration, vascular congestion, hemorrhage, increased collagen deposition and mast cell infiltration, compared to control rats. Montelukast preconditioning suppressed mast cell infiltration and inflammatory mediators to greater extent than MESNA. Also, montelukast enhanced autophagosomes formation in detrusor myocytes and up-regulated the autophagy-related proteins (beclin-1 & LC3-II), likely through inhibition of phosphatidylinositol-3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway. Montelukast preconditioning offset the up-regulation of transient receptor potential vanilloid 4 (TRPV4) in urothelial tissue of CPA-treated rats, to greater extent than MESNA. SIGNIFICANCE: These results demonstrate the uroprotective effect of montelukast on CPA-induced HC, which appears to be more superior to MESNA. These findings suggest that montelukast can emerge as a novel strategy to protect against CPA-induced urotoxicity.
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Acetatos/farmacologia , Autofagia , Ciclofosfamida/efeitos adversos , Ciclopropanos/farmacologia , Cistite/tratamento farmacológico , Hemorragia/tratamento farmacológico , Quinolinas/farmacologia , Sulfetos/farmacologia , Canais de Cátion TRPV/metabolismo , Animais , Apoptose , Cistite/induzido quimicamente , Hemorragia/induzido quimicamente , Inflamação , Masculino , Mastócitos/citologia , Mesna/farmacologia , Estresse Oxidativo , Fagocitose , Fagossomos/metabolismo , Ratos , Transdução de Sinais , Bexiga Urinária/metabolismo , Urotélio/metabolismoRESUMO
AIMS: Cyclophosphamide (CYP) is a potent anticancer agent with well-known cardiotoxicity that limits its clinical applications. Cilostazol is a vosodilating drug, showing a cardioprotective effect in some cardiac disorders; however its effect in CYP-induced cardiotoxicity is still uncertain. We investigated the effect of cilostazol against CYP-induced cardiotoxicity and the contribution of SIRT1 signaling. MATERIALS AND METHODS: 7 week-old male Wistar albino rats were treated with cilostazol (30 mg/kg/day, orally) in the absence or presence of SIRT1 inhibitor, EX-527 (5 mg/kg/day, IP) for 10 days and injected with CYP (200 mg/kg, IP) on the 7th day of the study. Age-matched rats were used as control group. On the 11th day, hearts were harvested for biochemical, immunoblotting and histological analyses. Markers of cardiac injury were assessed in plasma samples. KEY FINDINGS: CYP injection contributed to cardiac injury manifested as significant increases in plasma activities of heart enzymes and cardiac troponin I levels. Cilostazol attenuated cardiac injury and minimized the histological lesions in hearts of CYP-treated rats. Cilostazol induced 3 fold up-regulation of SIRT1 and promoted the antioxidant defense response through FoxO1-related mechanism in hearts of CYP-treated rats. Cilostazol suppressed the CYP-induced up-regulation of PARP1 and p53, and blocked the NF-kB p65-mediated inflammatory response in hearts of CYP-treated rats. All the beneficial effects of cilostazol were almost abolished by EX-527. SIGNIFICANCE: These data provided insights into the mechanism underlying the cardioprotective effect of cilostazol in CYP-treated rats through upregulation of SIRT1 signaling, suggesting that cilostazol might be a candidate modality for CYP-induced cardiotoxicity.
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Broncodilatadores/farmacologia , Cardiotoxicidade/prevenção & controle , Cilostazol/farmacologia , Ciclofosfamida/toxicidade , Inflamação/prevenção & controle , Mutagênicos/toxicidade , Sirtuína 1/metabolismo , Animais , Antioxidantes , Apoptose , Cardiotoxicidade/etiologia , Cardiotoxicidade/metabolismo , Cardiotoxicidade/patologia , Inflamação/etiologia , Inflamação/metabolismo , Inflamação/patologia , Masculino , Ratos , Ratos Wistar , Transdução de Sinais , Sirtuína 1/genéticaRESUMO
Stromal cell-derived factor-1α (SDF-1α) plays a key role in trafficking of stem cells and regeneration of injured tissue through interaction with its receptor, CXCR4. This study investigated the probable therapeutic effect of linagliptin (LG) against cisplatin (CP)-induced testicular injury and the underlying mechanisms. 12 week old male Sprague-Dawley rats were randomly assigned into 6 groups (n = 10 each) as follow: (i) Control, (ii) LG-treated control, (iii) CP-exposed rats, (iv) CP-exposed rats received LG, (v) CP-exposed rats received AMD3100, as CXCR4 antagonist, and (vi) CP-exposed rats received AMD3100 prior to LG. After 15 days, blood, testes and epididymides were collected for analyses. There were significant increases in both circulatory and testicular levels of SDF-1α in LG-treated rats. Conversely, higher levels of incretin hormones were found in serum but not in testicular tissue of rats, following LG therapy. CP injection significantly reduced body, testicular and epididymal weights of rats, and were restored by LG therapy. Treatment of CP-exposed rats with LG improved the deteriorated testicular architecture, reconstructed spermatogenesis, increased sperm count and quality, and normalized testosterone levels. LG therapy increased gene expression of Lin28a and Mvh, but did not alter the expressions of somatic-related genes. Additionally, LG therapy promoted germ cells survival and proliferation likely via activation of extracellular signal-regulated kinase1/2 (ERK1/2) signaling. These positive effects of LG therapy were almost blunted by administration of AMD3100. These results provided mechanistic insights into the ameliorative effect of LG on CP-induced testicular injury, through activation of SDF-1α/CXCR4 signaling pathway. Our findings suggest that LG can be a promising therapeutic candidate for CP-induced testicular injury.
Assuntos
Quimiocina CXCL12/imunologia , Cisplatino/efeitos adversos , Linagliptina/farmacologia , Doenças Testiculares , Testículo , Animais , Cisplatino/farmacologia , Masculino , Ratos , Ratos Sprague-Dawley , Doenças Testiculares/induzido quimicamente , Doenças Testiculares/imunologia , Doenças Testiculares/prevenção & controle , Testículo/imunologia , Testículo/lesõesRESUMO
High fat diet (HFD)-induced obesity adversely affects cardiac outcomes; however the effect of HFD consumption on myocardial remodeling and the underlying mechanisms are still elusive. This study aimed to examine the histological and molecular changes in cardiac tissue of HFD-fed rats. Eight-week old male Wistar rats were fed either HFD or normal chow diet for 16 weeks and then assessed for changes in metabolic and cardiac homeostasis (n = 10 each group). 16 weeks on HFD resulted in obesity, dyslipidemia and altered glucose tolerance but no hypertension. Histological examination of Masson's trichrome stained-cardiac sections revealed massive fibrotic changes, while immunoblotting analysis showed higher expressions of collagens I and III, and fibronectin in cardiac tissue of HFD-fed rats. The expressions of transforming growth factor beta1 and the phosphorylation of its downstream target, Smad3, were significantly increased in cardiac tissue of HFD-fed rats. Activation of endothelial-mesenchymal transition was promoted in hearts of HFD-fed rats, as evidenced by down-regulation of platelet endothelial cell adhesion molecule-1, while upregulation of α-smooth muscle actin and vimentin. Consumption of HFD induced dysregulation of AMP-activated protein kinase/glycogen synthase kinase-3 beta signaling in cardiac tissue of rats. This was coupled with down-regulation of nuclear factor erythroid-2-related factor 2 (Nrf2) and its downstream targets in cardiac tissue of HFD-fed rats, as well as enhanced the oxidative stress and inflammatory burden. These results demonstrate that moderate-term consumption of HFD can enhance oxidative stress, induce inflammation, and activate the fibrogenic pathways in cardiac tissue of rats which stimulate fibrotic remodeling. Our findings may implicate the dysregulation of Nrf2 signaling as a putative mechanism for this effect.
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Miocárdio/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Remodelação Ventricular/fisiologia , Proteínas Quinases Ativadas por AMP/metabolismo , Animais , Dieta Hiperlipídica/efeitos adversos , Dislipidemias/metabolismo , Fibrose , Glucose/metabolismo , Coração/fisiologia , Inflamação/metabolismo , Masculino , Fator 2 Relacionado a NF-E2/genética , Fator 2 Relacionado a NF-E2/fisiologia , Obesidade/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Fosforilação , Ratos , Ratos Wistar , Transdução de Sinais , Proteína Smad3/metabolismo , Fator de Crescimento Transformador beta1/metabolismoRESUMO
Diabetes induces time-dependent alterations in urinary bladders. Long-term diabetes causes an underactive bladder. However, the fundamental mechanisms are still elusive. This study aimed to examine the histological changes and the potential molecular pathways affected by long-term diabetes in the rat bladder. Diabetes was induced in 8-week-old male Lewis rats by streptozotocin, while age-matched control rats received citrate buffer only. Forty-four weeks after diabetes induction, bladders were harvested for histological and molecular analyses. The expressions of proteins related to fibrosis, apoptosis and oxidative stress as well as the cellular signaling pathway in the bladder were examined by immunoblotting. Histological examinations illustrated diabetes caused detrusor hypertrophy and fibrotic changes in the bladder. Immunoblotting analysis demonstrated higher collagen I but lower elastin expression in the bladder in diabetic rats. These were accompanied by an increase in the expression of transforming growth factor-beta1, along with the downregulation of matrix metalloptoteinase-1, and upregulation of tissue inhibitor of metalloproteinase-1. Diabetic rats showed an increase in nitrotyrosine, but decrease in nuclear factor erythroid-related factor 2 (Nrf2) levels in the bladder. Enhanced apoptotic signaling was observed, characterized by increased expression of Bcl-2-associated X protein (Bax), decreased expression of Bcl-2, in the diabetic bladder. The nerve growth factor level was decreased in the diabetic bladder. A significant suppression in the protein expressions of phosphorylated extracellular signal-regulated kinases 1/2 was found in diabetic bladders. This study demonstrated that long-term diabetes caused molecular changes that could promote fibrosis and apoptosis in the bladder. Oxidative stress may be involved in this context.
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Apoptose , Diabetes Mellitus Experimental/complicações , Fibrose/patologia , Estresse Oxidativo , Doenças da Bexiga Urinária/patologia , Bexiga Urinária/patologia , Animais , Biomarcadores/metabolismo , Fibrose/etiologia , Fibrose/metabolismo , Masculino , Ratos , Ratos Endogâmicos Lew , Transdução de Sinais , Bexiga Urinária/metabolismo , Doenças da Bexiga Urinária/etiologia , Doenças da Bexiga Urinária/metabolismoRESUMO
Bladder dysfunction in diabetes progresses gradually over time. However, the mechanisms of the development are not clear. We tested the hypothesis that oxidative stress plays a key role in the development of diabetic bladder dysfunction using an inducible smooth muscle (SM)-specific superoxide dismutase 2 (Sod2) gene knockout (SM-Sod2 KO) mouse model. Eight-week-old male Sod2lox/lox, SM-CreERT2(ki)Cre/+ mice and wild-type mice were assigned to diabetic or control groups. 4-Hydroxytamoxifen was injected into Sod2lox/lox, SM-CreERT2(ki)Cre/+ mice to activate CreERT2-mediated deletion of Sod2. Diabetes was induced by injection of streptozotocin, whereas control mice were injected with vehicle. Nine weeks later, bladder function was evaluated, and bladders were harvested for immunoblot analysis. Wild-type diabetic mice presented compensated bladder function along with increased nitrotyrosine and MnSOD in detrusor muscle. Induction of diabetes in SM-Sod2 KO mice caused deteriorated bladder function and even greater increases in nitrotyrosine compared with wild-type diabetic mice. Expression levels of apoptosis regulator Bax and cleaved caspase-3 were increased, but apoptosis regulator Bcl-2 expression was decreased in detrusor muscle of both diabetic groups, with more pronounced effects in SM-Sod2 KO diabetic mice. Our findings demonstrate that exaggerated oxidative stress can accelerate the development of bladder dysfunction in diabetic mice and the enhanced activation of apoptotic pathways in the bladder may be involved in the process.
Assuntos
Diabetes Mellitus Experimental/complicações , Diabetes Mellitus Experimental/patologia , Músculo Liso/metabolismo , Superóxido Dismutase/genética , Doenças da Bexiga Urinária/etiologia , Doenças da Bexiga Urinária/patologia , Animais , Peso Corporal , Caspase 3/metabolismo , Diabetes Mellitus Experimental/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Tamanho do Órgão , Estresse Oxidativo , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Superóxido Dismutase/metabolismo , Tirosina/análogos & derivados , Tirosina/metabolismo , Doenças da Bexiga Urinária/genética , Proteína X Associada a bcl-2/metabolismoRESUMO
Western diet (WD), rich in saturated fat and sugars, has become a risk factor for obesity and metabolic syndrome, however, its effect on endothelial function and vascular remodeling is not fully elucidated. Recent evidence suggests cross-talk between Rho kinase (ROCK) pathway and cardiovascular system. We aimed to investigate the effect of WD on aortic remodeling and the contribution of ROCK signaling. Eight week old male Sprague-Dawley rats were fed either standard chow diet (SD) or high fructose/ high-fat diet, typically as in WD. After 42 weeks, WD-fed rats showed hyperglycemia, dyslipidemia, and hypertension without marked weight gain, compared to SD-fed rats. Significant up-regulation of ROCK-1 and -2, along with a decline in eNOS expression were found in the aortic tissue of WD-fed rats. Additionally, WD-fed rats displayed oxidative stress and fibrosis in their aortic tissues versus controls. Our findings suggest that long-term feeding of WD contributes to endothelial dysfunction and aortic remodeling in adult male rats. ROCK activation seems to be involved in WD-related vascular disorders and may represent a promising therapeutic target.
Assuntos
Aorta/fisiopatologia , Dieta Ocidental/efeitos adversos , Endotélio/fisiopatologia , Remodelação Vascular , Quinases Associadas a rho/metabolismo , Animais , Dieta Hiperlipídica/efeitos adversos , Dislipidemias/etiologia , Hiperglicemia/etiologia , Hipertensão/etiologia , Masculino , Óxido Nítrico Sintase Tipo III/metabolismo , Estresse Oxidativo , Ratos , Ratos Sprague-Dawley , Transdução de SinaisRESUMO
The present study aimed to investigate the inhibitory effects of 10-dehydrogingerdione (10-DHGD) and pentoxifylline (PTX) either individually or in combined form on calcium deposition in high cholesterol diet (HCD)-fed rabbits as compared to atorvastatin (ATOR), and to clarify the underlying mechanisms. Three-months-old male New Zealand white rabbits received either normal chow or HCD for 12 weeks. The latter group was subdivided into five groups and concurrently treated either with vehicle (dyslipidemic control), ATOR, 10-DHGD, PTX or combined 10-DHGD and PTX. Blood samples and aortic tissue were collected for biochemical and histological analyses. HCD-fed rabbits displayed dyslipidemia, inflammation, atherosclerotic lesions, and calcium deposition in aortas as compared to normal group. This was associated with up-regulation of bone morphogenetic protein-2 (BMP-2), wingless-type MMTV integration site family 3A (Wnt3a) mRNA levels and osteopontin expression in their aortic tissue, along with higher serum alkaline phosphatase and osteocalcin levels. Furthermore, a marked decrease in osteoprotegerin, along with a significant increase in receptor activator of NF-κB(RANK) levels, was found in aortic tissue of dyslipidemic rabbits. 10-DHGD and PTX monotherapy significantly modulated the afore-mentioned calcification markers and attenuated aortic calcification to greater extent than ATOR. Combination of 10-DHGD and PTX exerted more anti-calcifying effect than either individual drug. Our findings suggested therapeutic roles of 10-DHGD and PTX against aortic calcium deposition in dyslipidemic rabbits, likely mediated by HDL-raising effect and attenuation of associated inflammation. Combination of 10-DHGD and PTX may represent a promising therapeutic strategy for aortic calcification associated with atherosclerosis.
Assuntos
Aorta , Cálcio/metabolismo , Colesterol/efeitos adversos , Gorduras na Dieta/efeitos adversos , Guaiacol/análogos & derivados , Pentoxifilina/farmacologia , Calcificação Vascular , Animais , Aorta/metabolismo , Aorta/patologia , Colesterol/farmacologia , Gorduras na Dieta/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Guaiacol/farmacologia , Masculino , Coelhos , Calcificação Vascular/induzido quimicamente , Calcificação Vascular/tratamento farmacológico , Calcificação Vascular/metabolismo , Calcificação Vascular/patologiaRESUMO
Policosanol (POL) is a hypocholesterolemic drug of natural origin and has been shown to reduce circulating levels of proprotein convertase subtilisin/kexin type 9 (PCSK9) in healthy participants. Recently, we have reported that POL can attenuate aortic calcification in diabetic dyslipidemic rats; however, the underlying mechanism is not fully elucidated. We aimed to investigate the effect of POL on aortic calcification and whether PCSK9 has a contributory role and also to examine whether the combination of POL with pentoxifylline (PTX) as anti-tumor necrosis factor α would offer additional benefits. Thirty adult male New Zealand rabbits weighing 1.5 to 2 kg were randomly assigned to 5 groups. One group received standard chow diet and served as normal control group (NC). The other 4 groups received 0.5% wt/wt cholesterol-rich diet for 12 weeks and concurrently treated with placebo, POL, PTX, or a combination of POL and PTX. Sera samples and aortic tissue were collected for biochemical measurements and histological assessment. Rabbits fed a cholesterol-rich diet demonstrated dyslipidemia, increased inflammatory state, and elevated serum levels of PCSK9, compared to the NC group. Aortic calcification was evident in dyslipidemic rabbits, represented by increased calcium deposition and osteopontin expression in aortic tissue, along with elevated serum levels of alkaline phosphatase and osteocalcin. Dyslipidemic rabbits showed a significant upregulation of wingless-type MMTV integration site family 3A and bone morphogenetic protein 2 genes in their aortic tissue. Policosanol significantly reduced circulating PCSK9 levels, suppressed calcification markers, and attenuated aortic calcification. Combination of POL with PTX alleviated aortic calcification to a greater extent than either monotherapy, which may be attributed to further suppression of PCSK9 and calcification markers. These findings suggested that POL exerted anticalcifying effect partly via inhibition of PCSK9. Combination of POL and PTX offered additional benefits and might represent a promising therapeutic option for aortic calcification.
Assuntos
Anticolesterolemiantes/farmacologia , Aorta/efeitos dos fármacos , Doenças da Aorta/prevenção & controle , Dislipidemias/tratamento farmacológico , Álcoois Graxos/farmacologia , Lipídeos/sangue , Inibidores de PCSK9 , Pentoxifilina/farmacologia , Inibidores de Serina Proteinase/farmacologia , Calcificação Vascular/prevenção & controle , Fosfatase Alcalina/sangue , Animais , Aorta/enzimologia , Aorta/patologia , Doenças da Aorta/sangue , Doenças da Aorta/enzimologia , Doenças da Aorta/patologia , Biomarcadores/sangue , Proteína Morfogenética Óssea 2/metabolismo , Modelos Animais de Doenças , Quimioterapia Combinada , Dislipidemias/sangue , Dislipidemias/enzimologia , Masculino , Osteocalcina/sangue , Osteopontina/metabolismo , Pró-Proteína Convertase 9/metabolismo , Coelhos , Transdução de Sinais/efeitos dos fármacos , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Fator de Necrose Tumoral alfa/metabolismo , Calcificação Vascular/sangue , Calcificação Vascular/enzimologia , Calcificação Vascular/patologia , Proteína Wnt3A/metabolismoRESUMO
AIMS: Diabetic bladder dysfunction (DBD) has been extensively studied in animal models of type 1 diabetes. We aimed to examine the functional and morphological alterations of the urinary bladder in a type 2 diabetes model, FVB(db/db) mice. METHODS: FVB(db/db) mice and age-matched FVB/NJ control mice were tested at either 12, 24 or 52weeks of age. Body weight, blood glucose and glycated hemoglobin (HbA1c) levels were measured. Bladder function was assessed by measurement of 24-h urination behavior and conscious cystometry. Bladder was harvested for Masson's Trichrome staining and morphometric analysis. RESULTS: The body weights of FVB(db/db) mice were twice as those of FVB/NJ control mice. The blood glucose and HbA1c levels were higher in FVB(db/db) mice at 12 and 24weeks, but not at 52weeks. A significant increase in the mean volume per void, but decrease in the voiding frequency, in FVB(db/db) mice was observed. Cystometry evaluation showed increased bladder capacity, voided volume, and peak micturition pressure in FVB(db/db) mice compared with FVB/NJ mice. Morphometric analysis revealed a significant increase in the areas of detrusor muscle and urothelium in FVB(db/db) mice. In addition, some FVB(db/db) mice, especially males at 12 and 24weeks, showed small-volume voiding during 24-h urination behavior measurement, and detrusor overactivity in the cystometry measurement. CONCLUSIONS: The FVB(db/db) mouse, displaying DBD characterized by not only increased bladder capacity, void volume, and micturition pressure, but also bladder overactivity, is a useful model to further investigate the mechanisms of type 2 diabetes-related bladder dysfunction.
Assuntos
Envelhecimento , Complicações do Diabetes/fisiopatologia , Diabetes Mellitus Tipo 2/complicações , Obesidade/complicações , Bexiga Urinária Hiperativa/complicações , Bexiga Urinária/fisiopatologia , Urotélio/fisiopatologia , Animais , Comportamento Animal , Glicemia , Complicações do Diabetes/sangue , Complicações do Diabetes/patologia , Ingestão de Líquidos , Feminino , Hemoglobinas Glicadas/análise , Masculino , Camundongos Endogâmicos , Camundongos Mutantes , Tamanho do Órgão , Caracteres Sexuais , Bexiga Urinária/patologia , Bexiga Urinária Hiperativa/sangue , Bexiga Urinária Hiperativa/patologia , Bexiga Urinária Hiperativa/fisiopatologia , Micção , Urotélio/patologia , Aumento de PesoRESUMO
Manganese superoxide dismutase (MnSOD) is considered a critical component of the antioxidant systems that protect against oxidative damage. We are interested in the role of oxidative stress in bladder detrusor smooth muscle (SM) in different disease states. In this study, we generated an inducible, SM-specific Sod2(-/-) mouse model to investigate the effects of MnSOD depletion on the function of the bladder. We crossbred floxed Sod2 (Sod2(lox/lox)) mice with mice containing heterozygous knock-in of a gene encoding a tamoxifen-activated Cre recombinase in the SM22α promoter locus [SM-CreER(T2)(ki)(Cre/+)]. We obtained Sod2(lox/lox),SM-CreER(T2)(ki)(Cre/+) mice and injected 8-wk-old males with 4-hydroxytamoxifen to induce Cre-mediated excision of the floxed Sod2 allele. Twelve weeks later, SM-specific deletion of Sod2 and depletion of MnSOD were confirmed by polymerase chain reaction, immunoblotting, and immunohistochemistry. SM-specific Sod2(-/-) mice exhibited normal growth with no gross abnormalities. A significant increase in nitrotyrosine concentration was found in bladder SM tissue of SM-specific Sod2(-/-) mice compared with both wild-type mice and Sod2(+/+), SM-CreER(T2)(ki)(Cre/+) mice treated with 4-hydroxytamoxifen. Assessment of 24-h micturition in SM-specific Sod2(-/-) mice revealed significantly higher voiding frequency compared with both wild-type and SM-specific Cre controls. Conscious cystometry revealed significantly shorter intercontraction intervals and lower functional bladder capacity in SM-specific Sod2(-/-) mice compared with wild-type mice. This novel model can be used for exploring the mechanistic role of oxidative stress in organs rich in SM in different pathological conditions.