RESUMO
OBJECTIVES: Cerebral stroke is a serious clinical condition in which oxidative stress, inflammation, necrosis, apoptosis, and autophagy play important roles in its pathogenesis. This study investigated the neuroprotective and healing effects of calcium dobesilate (CD) on cerebral hypoxia/reperfusion injury in rats. METHODS: Forty Wistar albino male rats, each weighing 300-350 g, were separated into the Control group (no surgery and no pharmacological agent was administered); Sham-A group (only surgery was performed); DBL-A group (surgery was performed and CD 100 mg/kg/day was administered intraperitoneally for 3 days); Sham-C group (only surgery was performed); and DBL-C group (surgery was performed and 100 mg/kg/day CD was administered intraperitoneally for 10 days). Under sedation anesthesia, the bilateral common carotid arteries of all rats except the Control group were clipped for 30 min. After 4 h, the CD was given to the relevant groups, and then, all subjects were euthanized at scheduled times. The brain of each animal was removed for histopathological (hematoxylin and eosin staining), immunohistochemical (beclin-1, anti-MHC class II and anti-CD-68 staining), and biochemical (TNF, IL-1ß, IL-6, caspase-3, GSH/GSSG, malondialdehyde, protein carbonyl, LC3II/LC3I, and beclin-1 levels) evaluations. RESULTS: It was observed that CD could reduce necrosis and mitigate polarization of microglia to the M1 phenotype, autophagy, free oxygen radicals, protein carbonylation, lipid peroxidation, IL-1ß, IL6, TNF, caspase-3, beclin-1, and LC3II/LC3I levels in acute and chronic periods of hypoxia/reperfusion injury. CONCLUSION: From these results, it was observed that CD treatment could reduce neuronal necrosis and create anti-inflammatory, anti-edema, anti-oxidant, anti-apoptotic, and anti-autophagic effects in hypoxia/reperfusion injury in rats.
Assuntos
Dobesilato de Cálcio , Hipóxia Encefálica , Traumatismo por Reperfusão , Ratos , Animais , Ratos Wistar , Caspase 3/metabolismo , Dobesilato de Cálcio/farmacologia , Dobesilato de Cálcio/uso terapêutico , Proteína Beclina-1 , Antioxidantes/uso terapêutico , Hipóxia , Necrose , Traumatismo por Reperfusão/metabolismoRESUMO
INTRODUCTION: Proinflammatory cytokines released from nerve endings and surrounding injured tissue after nerve damage can prolong the inflammation process, delay nerve healing or result in poor quality nerve healing. In this case, due to the loss of function in the muscles innervated by the damaged nerve, the patient may have neurological and functional difficulties which may reduce the patient's quality of life and create an economic burden. Although the attempts of many pharmacological agents to heal crush injury of peripheral nerves have been recorded in literature, a drug that can provide adequate recovery of the crushed nerve and can be applied in daily life has not been defined as yet. This study aimed to assess the effects of calcium dobesilate on sciatic nerve crush injury in a rat model. METHODS: A total of 26 male Wistar albino rats were separated into four groups as follows: CONTROL group (healthy subjects, n=6); SHAM group (crush injury was created, n=6); MP group (after created crush injury, methylprednisolone was administered, n=7); and CAD group (after created crush injury, calcium dobesilate was administered, n=7). A crush injury was created, then the electrophysiological findings and sciatic nerve functional index (SFI) were recorded before euthanasia. After the euthanasia of all the rats, samples of the crushed nerve and gastrocnemius muscle were evaluated histopathologically, immunohistochemically, and biochemically. RESULTS: Both pharmacological agents were histopathologically effective in axon regeneration and repair. Calcium dobesilate did not preserve total muscle mass but was seen to prevent atrophy microscopically. Immunohistochemistry and biochemistry results showed that calcium dobesilate and methylprednisolone had anti-inflammatory, anti-oxidant, anti-apoptotic, and anti-autophagic activity in the crushed sciatic nerve. Neither calcium dobesilate nor methylprednisolone improved the nerve conductance level. SFI values obtained on day 30 from the CAD group were numerically closer to the values of the healthy animals but not at a statistically significant level. CONCLUSION: The study results demonstrated that calcium dobesilate could suppress inflammatory processes and provide histopathological and functional improvements in the injured nerve in rats. Therefore, further clinical studies are recommended to investigate in detail the therapeutic effects of calcium dobesilate on peripheral nerve crush injury.
Assuntos
Dobesilato de Cálcio , Lesões por Esmagamento , Traumatismos dos Nervos Periféricos , Neuropatia Ciática , Animais , Ratos , Masculino , Dobesilato de Cálcio/farmacologia , Dobesilato de Cálcio/uso terapêutico , Axônios/patologia , Antioxidantes/farmacologia , Regeneração Nervosa/fisiologia , Qualidade de Vida , Ratos Wistar , Recuperação de Função Fisiológica , Nervo Isquiático/lesões , Lesões por Esmagamento/tratamento farmacológico , Metilprednisolona/farmacologia , Metilprednisolona/uso terapêutico , Anti-Inflamatórios/farmacologia , Citocinas , Neuropatia Ciática/tratamento farmacológico , Neuropatia Ciática/patologiaRESUMO
Calcium dobesilate (CaD) is used for the treatment of diabetic retinopathy and nephropathy. This agent exerts antioxidant effects. In the present study, we evaluated the protective effects of oral administration of CaD against hepatorenal damages in a mice model of aging induced by d-galactose (d-gal). We used 28 male albino mice, which equally and randomly were divided into four groups as follows: intact, aging (d-gal at the dose of 500 mg/kg, p.o.), aging + CaD 50 (d-gal plus CaD at the dose of 50 mg/kg), and aging + CaD 100 (d-gal plus CaD at the dose of 100 mg/kg, p.o.). All drugs were administered orally once a day for 42 days. The liver and kidney damages were evaluated by measuring mass indices, levels of serum creatinine and blood urea nitrogen, and activities of serum alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase and by histopathological evaluation. Moreover, hepatic and renal tissue oxidant/antioxidant markers (malondialdehyde, superoxide dismutase, catalase, and glutathione peroxidase) were measured. The results showed that d-gal treatment induced significant oxidative stress in the kidney and liver that was paralleled by dysfunctions and histological alterations of these organs. CaD significantly improved the liver and kidney indices, implemented functional capacity of the liver and kidney, as well as decreased oxidative stress enhancing antioxidative enzyme activities. CaD treatment also inhibited the development of histological alterations of both kidney and liver. CaD might represent a promising therapeutic agent for the attenuation of hepatorenal injuries induced by aging.
Assuntos
Dobesilato de Cálcio , Nefropatias , Animais , Antioxidantes/metabolismo , Dobesilato de Cálcio/metabolismo , Dobesilato de Cálcio/farmacologia , Galactose/toxicidade , Rim , Nefropatias/induzido quimicamente , Nefropatias/tratamento farmacológico , Nefropatias/prevenção & controle , Fígado , Masculino , Camundongos , Estresse Oxidativo , Superóxido Dismutase/metabolismoRESUMO
Acute kidney injury (AKI) is a serious complication of sepsis that increases mortality and the risk of progression to chronic kidney disease. Oxidative stress and apoptosis are reported to exert critical function in the pathogenesis of sepsis-associated AKI. Calcium dobesilate (CaD) was reported to play a protective role in renal diseases. Therefore, we explored the antioxidant effect and potential mechanism of CaD in lipopolysaccharide (LPS)-induced AKI in mice. We evaluated renal function (blood urea nitrogen (BUN) and serum creatinine (SCr)), histopathology, oxidative stress (superoxide dismutase (SOD) and malondialdehyde (MDA)), inflammation cytokines, and apoptosis in kidneys of mice. The effect of CaD on NF-κB signaling was evaluated by Western blot. Our findings showed that CaD alleviated renal dysfunction and kidney injury, and also reversed upregulated MDA concentration and reduced SOD enzyme activity in AKI mice. Moreover, LPS-induced inflammatory response was attenuated by CaD. CaD treatment also reduced the apoptosis evoked by LPS. Additionally, CaD downregulated phosphorylation of nuclear factor kappa B (NF-κB) signaling components in LPS mice. Conclusively, CaD alleviates renal dysfunction and inflammation by targeting NF-κB signaling in sepsis-associated AKI.
Assuntos
Injúria Renal Aguda/tratamento farmacológico , Dobesilato de Cálcio/farmacologia , NF-kappa B/metabolismo , Sepse/tratamento farmacológico , Transdução de Sinais/efeitos dos fármacos , Injúria Renal Aguda/etiologia , Injúria Renal Aguda/metabolismo , Animais , Inflamação/induzido quimicamente , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Lipopolissacarídeos/toxicidade , Masculino , Camundongos , Sepse/induzido quimicamente , Sepse/complicações , Sepse/metabolismoRESUMO
BACKGROUND: Serum creatinine is a widely used biomarker for evaluating renal function. Sarcosine oxidase enzymatic (SOE) analysis is currently the most widely used method for the detection of creatinine. This method was negatively interfered with by calcium dobesilate, causing pseudo-reduced results. The aim of this study was to explore a new method to alleviate the negative interference of this drug on creatinine detection. METHOD: We formulated eight drug concentrations and 12 creatinine concentrations from serum. The SOE method, the new method, and the Jaffe method were used for detection in five systems. Creatinine biases were analyzed under the conditions with or without the interference of calcium dobesilate, at consistent or inconsistent creatinine concentrations. Creatinine concentrations were also analyzed at three medical decision levels (MDLs). RESULTS: Calcium dobesilate had negative interference in creatinine SOE analysis. With the increase in calcium dobesilate concentrations, the negative bias increases. The new BG method showed an anti-negative interference effect. In the Roche system, the BG method reduced the negative bias from -71.11% to -16.7%. In the Abbott system, bias was reduced from -45.15% to -2.74%. In the Beckman system, the bias was reduced from -65.36% to -7.58%. In the Siemens system, the bias was reduced from -58.62% to -7.58%. In the Mindray system, the bias was reduced from -36.29% to -6.84%. CONCLUSION: The new method alleviated the negative interference of calcium dobesilate in creatinine SOE detection. The negative bias could be reduced from -60% or -70% to less than -20%.
Assuntos
Biomarcadores/sangue , Dobesilato de Cálcio/farmacologia , Ensaios Enzimáticos Clínicos/métodos , Creatinina/sangue , Nefropatias/diagnóstico , Sarcosina Oxidase/efeitos dos fármacos , Artefatos , Análise Química do Sangue , Hemostáticos/farmacologia , Humanos , Nefropatias/sangue , Testes de Função Renal , Sarcosina Oxidase/sangueRESUMO
BACKGROUND: Early acute kidney injury (AKI) predicts a high mortality rate in severely burned patients. However, the pathophysiology of early AKI induced by severe burn has not been well-defined. This study was designed to examine the protective effects of calcium dobesilate (CaD) against severe burn-induced early AKI in mice and explore the mechanism. METHODS: The shaved backs of mice were immersed in 100°C water for 10 s to make severe burn (40% of the total body surface area). CD-57 male mice were randomly divided into sham, burn, burn + vehicle, and burn + CaD groups. Renal function, reactive oxygen species generation, tubular necrosis, and phosphorylation of mitogen-activated protein kinase, protein kinase B (Akt), and nuclear factor (NF)-κB were measured at 24 and 48 h after the burn. Renal histology, ELISA, qRT-PCR, and Western blotting were performed on the renal tissue of mice to examine the effects and mechanisms at 24 and 48 h after the burn. RESULTS: Tubular damage, cast formation, and elevations of serum creatinine, BUN, and renal tissue kidney injury molecule 1 levels were all observed in the burned mice, and these were all alleviated in the mice with CaD treatment. In addition, the levels of oxidation-reduction potential and malondialdehyde were decreased, while the activities of the endogenous antioxidative enzymes were increased in the kidney tissues from the mice after CaD treatment. Furthermore, the activities of Akt, p38, extracellular sign-regulated kinase, Jun N-terminal kinase, and NF-κB signaling were increased in the kidney of burned mice and normalized after CaD treatment. CONCLUSION: This study has established, for the first time, the protective effect of CaD against early AKI in severely burned mice. CaD may exert its protective effect through alleviating oxidative stress, apoptosis, and inflammation, as well as modulating some signaling pathways in the kidney.
Assuntos
Injúria Renal Aguda/prevenção & controle , Queimaduras/complicações , Dobesilato de Cálcio/uso terapêutico , Injúria Renal Aguda/complicações , Injúria Renal Aguda/etiologia , Animais , Apoptose/efeitos dos fármacos , Dobesilato de Cálcio/farmacologia , Creatinina/sangue , Masculino , Camundongos , NF-kappa B/metabolismo , Oxirredução , Estresse Oxidativo/efeitos dos fármacos , Fosforilação , Índice de Gravidade de Doença , Transdução de Sinais/efeitos dos fármacosRESUMO
Cisplatin is one of the synthetic cancer medicines with nephrotoxicity being one of its major side effects. Past research shows that calcium dobesilate (CaD), as a vascular protective agent in diabetic retinopathy, has antioxidant properties. Thus, this study aims to evaluate the protective effects of CaD in cisplatin-induced nephrotoxicity in mice. A many as 28 mice, in the present experimental research, were randomly distributed into four groups, including control, cisplatin (the intraperitoneal administration of 20 mg/kg cisplatin only on the first day of the experiment), cisplatin + CaD 50 (cisplatin with the oral administration of 50 mg/kg CaD), and cisplatin + CaD 100 (cisplatin with the oral administration of 100 mg/kg CaD). The treated groups received CaD by oral gavage for 4 constitutive days. On the fifth day, the mice were sacrificed, and some biochemical (serum levels of Cr and BUN, renal tissue levels of MDA, and renal activities of SOD and GPx) and pathological parameters were evaluated. Based on the results, there was a significant decrease in the renal SOD and GPx activities; in contrast, there was a significant increase in the BUN, Cr, and renal MDA levels following administering cisplatin. However, the CaD treatment (100 mg/kg) significantly attenuated these alterations. In addition, the kidney's histological examination of kidneys confirmed the nephroprotective effects of CaD. The findings proved the protective impact of CaD on cisplatin-induced nephrotoxicity by an improvement in the oxidative stress factors.
Assuntos
Antineoplásicos/toxicidade , Dobesilato de Cálcio/uso terapêutico , Cisplatino/toxicidade , Nefropatias/tratamento farmacológico , Substâncias Protetoras/uso terapêutico , Animais , Nitrogênio da Ureia Sanguínea , Dobesilato de Cálcio/farmacologia , Creatinina/sangue , Glutationa Peroxidase/metabolismo , Rim/efeitos dos fármacos , Rim/metabolismo , Rim/patologia , Nefropatias/induzido quimicamente , Nefropatias/metabolismo , Nefropatias/patologia , Masculino , Malondialdeído/metabolismo , Camundongos , Estresse Oxidativo/efeitos dos fármacos , Substâncias Protetoras/farmacologia , Superóxido Dismutase/metabolismoRESUMO
Calcium dobesilate (Cad), a protective agent, protects against microvascular damage, and diseases such as diabetic retinopathy and diabetic nephropathy. However, these vascular protective effects have not been demonstrated in chronic kidney disease (CKD). In this study, we aimed to determine the ability of Cad to protect against renal interstitial fibrosis induced by unilateral ureteral obstruction (UUO) and identify the underlying therapeutic mechanisms of Cad during hypoxia/serum deprivation (H/SD) in human umbilical vein endothelial cells (HUVECs). A total of 36 male mice were randomly assigned into 3 groups (12 mice in each group): the Sham-operated group (Sham), the saline solution-treated UUO mice group (UUO), and the Cad administration (intragastrically) group (Cad). The mice in Cad group were administered Cad (100 mg/kg) daily by oral gavage and slaughtered on the 7th and 14th days post-surgery. Six mice from each group were sacrificed by sodium pentobarbital injection on the 7th and 14th day after surgery. Tissue hypoxia, cell apoptosis and fibrotic lesions were detected by Immunostaining and Western blot. Peritubular capillaries (PTCs) injury was measured by a novel technique of fluorescent microangiography (FMA). Endothelial cell-to-mesenchymal transition (EndMT) were identified by immunofluorescence and Western blot. HUVECs proliferation was measured via Cell Counting Kit8 assays and Edu staining. Sirt1 and its downstream gene in Cad regulation of endothelial were detected. Hematoxylin-eosin (HE), Masson-trichrome stains and Histological findings showed that Cad administration markedly reduced hypoxia and renal interstitial fibrosis at each time point in UUO. Meanwhile, Cad protect against EndMT process of PTCs by increasing CD31 expression and decreasing α-smooth muscle actin and fibronectin expression. in vitro studies showed that there was a proliferative response of the HUVECs incubated with Cad (10 µM) in H/SD. Sirt1 was suppressed after small interfering RNA (siRNA) was transfected in HUVECs. Mechanistically, Cad enhanced Sirt1 signaling, which was accompanied by increased levels of p53 acetylation (ac-p53). Meanwhile, protein expression of Bcl-2, and VE-cadherin were downregulated, Bax, and α-SMA were upregulated. In summary, the therapeutic effect of Cad in obstructive nephropathy were likely through suppressing EndMT progression and promoting anti-apoptotic effects after via activating the Sirt1/p53 signaling pathway.
Assuntos
Dobesilato de Cálcio/farmacologia , Nefropatias/prevenção & controle , Sirtuína 1/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Animais , Apoptose/efeitos dos fármacos , Capilares/efeitos dos fármacos , Capilares/patologia , Modelos Animais de Doenças , Progressão da Doença , Fibrose , Hemostáticos/farmacologia , Células Endoteliais da Veia Umbilical Humana , Humanos , Nefropatias/fisiopatologia , Túbulos Renais/irrigação sanguínea , Masculino , Camundongos , Camundongos Endogâmicos C57BLRESUMO
Inhibiting vascular endothelial growth factor (VEGF) is a therapeutic option in diabetic microangiopathy. However, VEGF is needed at physiological concentrations to maintain glomerular integrity; complete VEGF blockade has deleterious effects on glomerular structure and function. Anti-VEGF therapy in diabetes raises the challenge of reducing VEGF-induced pathology without accelerating endothelial cell injury. Heparan sulfate (HS) act as a co-receptor for VEGF. Calcium dobesilate (CaD) is a small molecule with vasoprotective properties that has been used for the treatment of diabetic microangiopathy. Preliminary evidence suggests that CaD interferes with HS binding sites of fibroblast growth factor. We therefore tested the hypotheses that (1) CaD inhibits VEGF signaling in endothelial cells, (2) that this effect is mediated via interference between CaD and HS, and (3) that CaD ameliorates diabetic nephropathy in a streptozotocin-induced diabetic mouse model by VEGF inhibition. We found that CaD significantly inhibited VEGF165-induced endothelial cell migration, proliferation, and permeability. CaD significantly inhibited VEGF165-induced phosphorylation of VEGFR-2 and suppressed the activity of VEGFR-2 mediated signaling cascades. The effects of CaD in vitro were abrogated by heparin, suggesting the involvement of heparin-like domain in the interaction with CaD. In addition, VEGF121, an isoform which does not bind to heparin, was not inhibited by CaD. Using the proximity ligation approach, we detected inhibition of interaction in situ between HS and VEGF and between VEGF and VEGFR-2. Moreover, CaD reduced VEGF signaling in mice diabetic kidneys and ameliorated diabetic nephropathy and neuropathy, suggesting CaD as a VEGF inhibitor without the negative effects of complete VEGF blockade and therefore could be useful as a strategy in treating diabetic nephropathy.
Assuntos
Dobesilato de Cálcio/farmacologia , Diabetes Mellitus Experimental/tratamento farmacológico , Nefropatias Diabéticas/tratamento farmacológico , Fator A de Crescimento do Endotélio Vascular/genética , Animais , Sítios de Ligação/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Diabetes Mellitus Experimental/genética , Diabetes Mellitus Experimental/patologia , Nefropatias Diabéticas/genética , Nefropatias Diabéticas/patologia , Células Endoteliais/efeitos dos fármacos , Heparitina Sulfato/metabolismo , Humanos , Cinética , Camundongos , Camundongos Endogâmicos NOD/genética , Camundongos Endogâmicos NOD/crescimento & desenvolvimento , Ligação Proteica/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Fator A de Crescimento do Endotélio Vascular/antagonistas & inibidores , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/genéticaRESUMO
Micro-vascular diseases and its associated complications continue to be a significant health problem worldwide. Vascular lesions from microvascular involvement lead to impaired blood flow and contribute to damage and dysfunction of one or more target organs, that is, the heart, kidneys, eyes, and nervous system. Calcium Dobesilate Drug (CAD) is an established vasoactive and angioprotective drug that has shown a unique, multitarget mode of action in several experimental studies and in different animal models of diabetic microvascular complications. CAD has been widely used as an antioxidant and a vascular protective agent. At present, the application of Calcium Dobesilate is mainly related to Micro-vascular damage-related diseases, such as diabetic retinopathy (DR) and diabetic nephropathy (DN), and it is found to significantly improve the related symptoms. Its beneficial effects make it an attractive therapeutic compound especially in the early stages of these diseases. Scholars at home and abroad have studied the effectiveness, safety, and mechanisms of the related diseases, furthermore, the subjects involved patients and animal models, they have found some new clinical effects of this medicine. This paper makes a brief summary of a research progress of clinical application about Vascular injury related diseases and other aspects.
Assuntos
Dobesilato de Cálcio/metabolismo , Dobesilato de Cálcio/farmacologia , Microvasos/efeitos dos fármacos , Animais , Antioxidantes , Nefropatias Diabéticas/tratamento farmacológico , Retinopatia Diabética/tratamento farmacológico , Humanos , Estresse Oxidativo , Doenças Vasculares/tratamento farmacológico , Doenças Vasculares/metabolismoRESUMO
Purpose/Aim of the study: Inflammation and oxidative stress are two significant factors affecting the degree of liver damage in obstructive jaundice. The aim of this study was to evaluate the effect of calcium dobesilate (CaDob), an effective antioxidant and anti-inflammatory drug, on damage to liver caused by experimental obstructive jaundice. MATERIALS AND METHODS: 30 rats in total were randomly placed into three groups, each group consisting of 10 rats. The sham group (Group 1) only received solely laparotomy. In the control group (Group 2), ligation was applied to the biliary tract and no treatment was implemented. In the CaDob group (Group 3), following ligation of the biliary tract, 100 mg/kg/day CaDob was implemented via an orogastric tube for a 10-day period. Liver tissue and blood samples were taken for histopathological and biochemical examination. RESULTS: The CaDob group had significantly lower test values for serum liver functions when compared to the control group. Statistically lower levels of tissue malondialdehyde (MDA) and fluorescent oxidation products (FOP) were detected in the CaDob group, and the CaDob group had significantly higher levels of sulfydryl (SH) than the control group. Histopathological scores in the CaDob group were found out to be statistically less than the scores the control group received (p < 0.05). CONCLUSIONS: CaDob treatment repaired the histpatological changes induced by bile duct ligation. The hepatoprotective effects of CaDob can be associated with its antioxidant properties of the drug.
Assuntos
Antioxidantes/farmacologia , Dobesilato de Cálcio/farmacologia , Icterícia Obstrutiva/tratamento farmacológico , Fígado/efeitos dos fármacos , Animais , Antioxidantes/uso terapêutico , Dobesilato de Cálcio/uso terapêutico , Modelos Animais de Doenças , Avaliação Pré-Clínica de Medicamentos , Feminino , Humanos , Icterícia Obstrutiva/etiologia , Icterícia Obstrutiva/patologia , Fígado/patologia , Fígado/fisiopatologia , Testes de Função Hepática , Estresse Oxidativo/efeitos dos fármacos , Ratos , Ratos Wistar , Resultado do TratamentoRESUMO
BACKGROUND: This study investigates the protective effect of calcium dobesilate (CaDob), an effective antioxidant and anti-inflammatory drug, on experimental liver ischemia-reperfusion injury (IRI). METHODS: Forty rats were divided into four groups. In Group 1, (sham), only hepatic pedicle was induced. In Group 2 (control), hepatic pedicle was reperfused for 90 min after being clamped for 60 min. No treatment was given in Group 1 and 2. In Group 3 (perioperative CaDob), 100 mg/kg CaDob was given 2 hours prior to the operation in which hepatic pedicle was reperfused for 90 min following a 60-min clamp. In Group 4 (preoperative CaDob), after 100 mg/kg/day CaDob was given for 10 days before the operation, hepatic pedicle was clamped for 60 min and reperfused for 90 minutes. At the end of these procedures, blood and liver tissue samples were collected for biochemical and histopathological assessment. RESULTS: Liver function tests and tissue oxidative stress parameters were significantly lower in the preoperative and perioperative treatment groups than the control group. Furthermore, it was observed that histopathological injury in the control group significantly decreased in both perioperative and preoperative treatment groups. CONCLUSION: Calcium dobesilate demonstrated a significant hepatoprotective effect in terms of its antioxidant and anti-inflammatory effects.
Assuntos
Antioxidantes/farmacologia , Dobesilato de Cálcio/farmacologia , Hepatopatias/metabolismo , Fígado/efeitos dos fármacos , Traumatismo por Reperfusão/metabolismo , Animais , Estresse Oxidativo/efeitos dos fármacos , RatosRESUMO
BACKGROUND: We reported that calcium dobesilate, a vasoprotective agent mainly used for diabetic retinopathy (DR), negatively interferes with glycated albumin (GA) assays involving enzymatic methods. METHODS: A calcium dobesilate standard was added to 3serum pools in vitro to prepare concentration-response series according to Clinical and Laboratory Standards Institute EP7-A2 guidelines. Percentage deviation between each drug concentration and the drug-free sample was calculated for 6 commercially available GA assays. The acceptable limit of deviation for GA was ±5.61%. For in vivo analyses, changes in serum concentrations of GA and calcium dobesilate were monitored in eight healthy participants before and after oral calcium dobesilate administration. RESULTS: At 16⯵g/ml calcium dobesilate, within the therapeutic range, the percentage deviations for Asahi Kasei, Maccura, Leadman, Homa, and Medicalsystem assays were -8.7% to -49.7%, -2.0% to -47.7%, and -10.1% to -35.7% for low-, medium- and high-GA level interference pools, respectively, exhibiting dose-dependent negative interference. In vivo, calcium dobesilate ingestion was associated with statistically significant, falsely decreased measurements in 5 GA assays, 2â¯h after daily 500â¯mg administration. CONCLUSIONS: Calcium dobesilate ingestion was associated with erroneously low measurements in 5 GA assays. The degree of interference varied greatly among the assays examined.
Assuntos
Artefatos , Análise Química do Sangue/métodos , Dobesilato de Cálcio/farmacologia , Retinopatia Diabética/tratamento farmacológico , Enzimas/metabolismo , Albumina Sérica/análise , Administração Oral , Adulto , Dobesilato de Cálcio/administração & dosagem , Dobesilato de Cálcio/uso terapêutico , Retinopatia Diabética/sangue , Feminino , Produtos Finais de Glicação Avançada , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem , Albumina Sérica GlicadaRESUMO
Diabetic kidney disease (DKD) is a leading cause of endstage renal disease. However, the pathogenesis of DKD remains unclear, and no effective treatments for the disease are available. Thus, there is an urgent need to elucidate the pathogenic mechanisms of DKD and to develop more effective therapies for this disease. Human umbilical vein endothelial cells (HUVECs) were cultured using different Dglucose concentrations to determine the effect of high glucose (HG) on the cells. Alternatively, HUVECs were incubated with 100 µmol/l calcium dobesilate (CaD) to detect its effects. The authors subsequently measured HUVEC proliferation via cell counting kit8 assays. In addition, HUVEC angiogenesis was investigated via migration assays and fluorescein isothiocyanate (FITC)labelled bovine serum albumin (BSA) permeability assays. The content or distribution of markers of endothelial dysfunction [vascular endothelial growth factor (VEGF), VEGF receptor (R) and endocan) or inflammation [intercellular adhesion molecule (ICAM)1, monocyte chemotactic protein (MCP)1 and pentraxinrelated protein (PTX3)] was evaluated via reverse transcriptionquantitative polymerase chain reaction and western blotting. HG treatment induced increased in VEGF, VEGFR, endocan, ICAM1, MCP1 and PTX3 mRNA and protein expression in HUVECs. HG treatment for 24 to 48 h increased cell proliferation in a timedependent manner, but the cell proliferation rate was decreased at 72 h of HG treatment. Conversely, CaD inhibited abnormal cell proliferation. HG treatment also significantly enhanced HVUEC migration compared to the control treatment. In contrast, CaD treatment partially inhibited HUVEC migration compared to HG exposure. HGtreated HUVECs exhibited increased FITCBSA permeability compared to control cells cultured in medium alone; however, CaD application prevented the HGinduced increase in FITCBSA permeability and suppressed HGinduced overexpression of endothelial markers (VEGF, VEGFR2, endocan) and inflammation markers (ICAM1, MCP1, PTX3) in HUVECs. CaD has angioprotective properties and protects endothelial cells partly by ameliorating HGinduced inflammation. The current results demonstrated the potential applicability of CaD to the treatment of diabetic nephropathy, particularly during the early stages of this disease.
Assuntos
Dobesilato de Cálcio/uso terapêutico , Diabetes Mellitus/tratamento farmacológico , Diabetes Mellitus/fisiopatologia , Endotélio Vascular/patologia , Endotélio Vascular/fisiopatologia , Inflamação/patologia , Biomarcadores/metabolismo , Proteína C-Reativa/metabolismo , Dobesilato de Cálcio/farmacologia , Permeabilidade da Membrana Celular/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Quimiocina CCL2/metabolismo , Diabetes Mellitus/patologia , Endotélio Vascular/efeitos dos fármacos , Glucose/toxicidade , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Mediadores da Inflamação/metabolismo , Molécula 1 de Adesão Intercelular/metabolismo , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/metabolismo , Proteoglicanas/genética , Proteoglicanas/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Componente Amiloide P Sérico/metabolismo , Fatores de Tempo , Regulação para Cima/efeitos dos fármacos , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismo , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/genética , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismoRESUMO
Calcium dobesilate (CaD) has been prescribed to some patients in the early stages of diabetic retinopathy to delay its progression. We previously reported that the treatment of diabetic animals (4 weeks of diabetes) with CaD, during the last 10 days of diabetes, prevents blood-retinal barrier breakdown. Here, we aimed to investigate whether later treatment of diabetic rats with CaD would reverse inflammatory processes in the retina. Diabetes was induced with streptozotocin, and 6 weeks after diabetes onset, CaD (100 mg/kg/day) was administered for 2 weeks. The treatment with CaD significantly increased glial fibrillary acidic protein (GFAP) levels in the retina of nondiabetic animals (138.6 ± 12.8% of control) and enhanced the diabetes-induced increase in GFAP levels (174.8 ± 5.6% of control). In addition, CaD prevented the increase in mRNA and protein expression of tumor necrosis factor and interleukin-1ß, as well as the formation of oxidized carbonyl residues and the increase in nitrotyrosine immunoreactivity, particularly in the ganglion cell layer of diabetic animals. We demonstrate that the treatment of diabetic animals with CaD can reverse the established proinflammatory processes in the retina. These beneficial effects appear to be attributed, at least partially, to the antioxidant properties of CaD.
Assuntos
Dobesilato de Cálcio/farmacologia , Diabetes Mellitus Experimental , Diabetes Mellitus Tipo 1/complicações , Retinopatia Diabética/prevenção & controle , Inflamação/prevenção & controle , Estresse Oxidativo/efeitos dos fármacos , Retina/patologia , Animais , Apoptose/efeitos dos fármacos , Barreira Hematorretiniana/efeitos dos fármacos , Diabetes Mellitus Tipo 1/metabolismo , Retinopatia Diabética/diagnóstico , Retinopatia Diabética/metabolismo , Hemostáticos/farmacologia , Inflamação/metabolismo , Inflamação/patologia , Masculino , Ratos , Ratos Wistar , Retina/metabolismoRESUMO
AIM: Calcium dobesilate (CaD) is beneficial in early stages of diabetic retinopathy (DR), but its mechanisms of action remains to be elucidated. The aim was to investigate the effect of CaD on proinflammatory cytokines and oxidative stress. METHODS: db/db mice were randomly assigned to daily oral treatment with CaD (200mg/kg/day) or vehicle for 15days. Biomarkers of oxidative stress (dihydroethidium, malondialdehyde), NF-κB, and proinflammatory cytokines (IL-1ß, IL-6, IL-8, TNF-α, MCP-1) were examined in the retina by immunohistochemical analysis. Cultures of human retinal endothelial cells (HRECs) were used for complementary experiments. RESULTS: CaD significantly reduced the biomarkers of oxidative stress in the retina of db/db mice. In addition, CaD prevented the increase of NF-κB, IL-6, IL-8, TNF-α and MCP-1 induced by diabetes. CaD inhibited the activation of NF-kß induced by IL-1ß by preventing IKKB-α phosphorylation in HRECs and reduced the upregulation of IL-6 and IL-18 induced by TNF-α in a dose-dependent manner. CONCLUSION: Our results suggest that antioxidant and antiinflammatory effects are crucial in accounting for the effectiveness of CaD for treating DR.
Assuntos
Anti-Inflamatórios não Esteroides/uso terapêutico , Dobesilato de Cálcio/uso terapêutico , Retinopatia Diabética/prevenção & controle , Fármacos Neuroprotetores/uso terapêutico , Estresse Oxidativo/efeitos dos fármacos , Retina/efeitos dos fármacos , Retinite/prevenção & controle , Animais , Anti-Inflamatórios não Esteroides/farmacologia , Antioxidantes/farmacologia , Antioxidantes/uso terapêutico , Biomarcadores/metabolismo , Dobesilato de Cálcio/farmacologia , Células Cultivadas , Cruzamentos Genéticos , Citocinas/antagonistas & inibidores , Citocinas/metabolismo , Diabetes Mellitus Tipo 2/complicações , Diabetes Mellitus Tipo 2/tratamento farmacológico , Retinopatia Diabética/imunologia , Retinopatia Diabética/metabolismo , Retinopatia Diabética/patologia , Humanos , Quinase I-kappa B/metabolismo , Masculino , Camundongos Mutantes , Camundongos Transgênicos , Fosforilação/efeitos dos fármacos , Processamento de Proteína Pós-Traducional/efeitos dos fármacos , Distribuição Aleatória , Retina/imunologia , Retina/metabolismo , Retina/patologia , Retinite/complicações , Retinite/imunologia , Retinite/metabolismoRESUMO
PURPOSE: The mechanisms involved in the reported beneficial effects of Calcium dobesilate monohydrate (CaD) for the treatment of diabetic retinopathy (DR) remain to be elucidated. The main aim of the present study is to examine whether CaD prevents early events in the pathogenesis of DR such as neurodegeneration and vascular leakage. In addition, putative mediators of both neurodegeneration (glutamate/GLAST, ET-1/ETB receptor) and early microvascular impairment (ET-1/ETA receptor, oxidative stress, VEGF, and the PKC-delta-p38 MAPK pathway) have been examined. METHODS: Diabetic (db/db) mice were randomly assigned to daily oral treatment with CaD (200 mg/Kg/day) (n = 12) or vehicle (n = 12) for 14 days. In addition, 12 non-diabetic (db/+) mice matched by age were used as the control group. Functional abnormalities were assessed by electroretinography. Neurodegeneration and microvascular abnormalities were evaluated by immunohistochemistry and Western blot. Glutamate was determined by HPLC. RESULTS: CaD significantly decreased glial activation and apoptosis and produced a significant improvement in the electroretinogram parameters. Mechanistically, CaD prevented the diabetes-induced up-regulation of ET-1 and its cognate receptors (ETA-R and ETB-R), which are involved in microvascular impairment and neurodegeneration, respectively. In addition, treatment with CaD downregulated GLAST, the main glutamate transporter, and accordingly prevented the increase in glutamate. Finally, CaD prevented oxidative stress, and the upregulation of VEGF and PKC delta-p38 MAPK pathway induced by diabetes, thus resulting in a significant reduction in vascular leakage. CONCLUSIONS: Our findings demonstrate for the first time that CaD exerts neuroprotection in an experimental model of DR. In addition, we provide first evidence that CaD prevents the overexpression of ET-1 and its receptors in the diabetic retina. These beneficial effects on the neurovascular unit could pave the way for clinical trials addressed to confirm the effectiveness of CaD in very early stages of DR.
Assuntos
Dobesilato de Cálcio/farmacologia , Diabetes Mellitus Experimental , Diabetes Mellitus Tipo 2/complicações , Retinopatia Diabética/prevenção & controle , Estresse Oxidativo/genética , Retina/metabolismo , Animais , Apoptose/efeitos dos fármacos , Western Blotting , Sobrevivência Celular , Células Cultivadas , Diabetes Mellitus Tipo 2/metabolismo , Retinopatia Diabética/diagnóstico , Retinopatia Diabética/etiologia , Eletrorretinografia , Ensaio de Imunoadsorção Enzimática , Regulação da Expressão Gênica , Hemostáticos/farmacologia , Humanos , Marcação In Situ das Extremidades Cortadas , Masculino , Camundongos , Microscopia de Fluorescência , Estresse Oxidativo/efeitos dos fármacos , RNA/genética , Reação em Cadeia da Polimerase em Tempo Real , Retina/efeitos dos fármacos , Retina/patologia , Epitélio Pigmentado da Retina/metabolismo , Epitélio Pigmentado da Retina/patologia , Fator A de Crescimento do Endotélio Vascular/biossíntese , Fator A de Crescimento do Endotélio Vascular/genéticaRESUMO
Apoptosis of renal proximal tubular epithelial cells (PTECs) plays a vital role in the pathogenesis and progression of diabetic kidney disease (DKD). Calcium dobesilate is a vascular protective compound used for treatment of diabetic retinopathy and chronic venous insufficiency. The aim of this study was to determine whether calcium dobesilate can protect PTECs from glucose-induced apoptosis and the potential mechanism of this effect. It is indicated that high glucose promoted abnormal apoptosis of HK2 cells, which was inhibited by treatment of calcium dobesilate, while Bim expression decreased in response to calcium dobesilate in high-glucose-treated HK2 cells. These findings confirmed the therapeutic effects of calcium dobesilate on DKD and emphasized the importance of it as a potentially crucial drug in treatment of DKD.
Assuntos
Apoptose , Proteína 11 Semelhante a Bcl-2/metabolismo , Dobesilato de Cálcio/farmacologia , Túbulos Renais Proximais/efeitos dos fármacos , Dobesilato de Cálcio/uso terapêutico , Linhagem Celular , Nefropatias Diabéticas/prevenção & controle , Células Epiteliais/efeitos dos fármacos , Glucose/farmacologia , Humanos , Túbulos Renais Proximais/citologiaRESUMO
OBJECTIVE: In this study, we investigated whether the administration of calcium dobesilate (CD) affects oxidative stress markers and histopathological outcomes in a rat model of intestinal ischemia-reperfusion (IR) injury. MATERIALS AND METHODS: This study was conducted with 30 male Wistar rats. The rats were randomly assigned to three groups as follows: a sham group (n = 10), an IR group (n = 10), and an IR + CD group (n = 10). In the sham group, superior mesenteric artery (SMA) dissection alone was performed during laparotomy. In the IR group, the procedure included SMA occlusion for 60 min, followed by reperfusion for 60 min. In the IR + CD group, CD (100 mg/kg/day) was additionally given for two days before laparotomy by intragastric lavage. In all the rats, 2 ml of blood were drawn, and an ileal segment (approximately 2 cm in size) was removed to evaluate oxidative stress markers. The ileal segment removed was divided into two pieces, and one piece was reserved for histopathological evaluation. RESULTS: Compared to the other groups, both serum and tissue oxidative stress indices were lower in the IR + CD group. The decrease was due to CD increasing the total antioxidant capacity. Moreover, the histological analysis showed that CD reduced tissue injury. CONCLUSIONS: CD may exert a protective effect against intestinal IR injury by increasing antioxidant capacity.