Efeito da curcumina e carotenoides em iogurte sobre biomarcadores fisio-metabólicos e de estresse oxidativo em modelos de diabetes mellitus tipo 1 e de obesidade/resistência insulínica
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Data
2016-07-26
Autores
Assis, Renata Pires de [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
As mudanças no estilo de vida e na estrutura dietética da população atual levam a uma ingestão calórica excessiva e ao comportamento sedentário, que em conjunto pode explicar a epidemia de obesidade e diabetes mellitus (DM). O diabetes mellitus é definido como um grupo de doenças metabólicas caracterizado por hiperglicemia, resultante da deficiência na produção e secreção de insulina pelas células beta pancreáticas e/ou resistência à ação da insulina em tecidos-alvo. A manutenção da hiperglicemia por longos períodos é um dos principais fatores envolvidos no estabelecimento do estresse oxidativo, condição esta que participa no desenvolvimento de vários prejuízos do DM, em especial as complicações micro e macrovasculares. Diversos trabalhos demonstram os efeitos de produtos naturais na melhoria da hiperglicemia, estresse oxidativo, status inflamatório e nas complicações do DM. Também tem sido explorado o uso de matrizes alimentares como veículos para ativos naturais, como por exemplo, o iogurte. Neste contexto, utilizamos dois modelos experimentais para avaliar os efeitos dos tratamentos com curcumina, bixina ou licopeno incorporados ao iogurte, isolados ou em associação: i) DM tipo 1 induzido com estreptozotocina (STZ); ii) obesidade/resistência insulínica induzida com dieta high-fat (HF). Experimento I: No modelo de DM induzido com STZ, foram utilizados ratos Wistar machos, normais (N) e diabéticos (D, 40 mg/kg STZ, i.v.), tratados durante 50 dias, distribuídos nos seguintes grupos: NIOG e DIOG, tratados com iogurte; DINS, tratados com insulina (4 U/dia); DC, tratados com 90 mg/kg de curcumina; DB, tratados com 5,5 mg/kg de bixina; DCB, tratados com 90 mg/kg de curcumina e 5,5 mg/kg de bixina; DL, tratados com 45 mg/kg de licopeno; DCL, tratados com 90 mg/kg de curcumina e 45 mg/kg de licopeno. Todos os ativos naturais foram administrados em iogurte. Foram analisados os seguintes parâmetros: i) fisiológicos: peso corporal, ingestão alimentar e hídrica, volume urinário, massas dos tecidos adiposos epididimal e retroperitoneal e dos músculos esqueléticos soleus e extensor digitorum longus (EDL); ii) biomarcadores bioquímicos plasmáticos, urinários e teciduais: glicemia, colesterol total, colesterol-HDL, triacilgliceróis (TAG), alanina aminotransferase (ALT), aspartato aminotransferase (AST), fosfatase alcalina (ALP), glicosúria, proteinúria e ureia urinária, conteúdo de glicogênio hepático; iii) biomarcadores plasmáticos e teciduais relacionados ao estresse oxidativo: substâncias reativas ao ácido tiobarbitúrico (malondialdeído, MDA), níveis de LDL-oxidada (LDL-ox), níveis de proteínas carboniladas (PCO), níveis de glutationa reduzida (GSH), atividades da superóxido dismutase (SOD), catalase (CAT) e glutationa peroxidase (GSH-Px). Como esperado, os animais DIOG apresentaram piora em todos os biomarcadores analisados, durante todo o período experimental, enquanto que o tratamento com insulina (DINS) foi eficaz na melhoria de todos os biomarcadores alterados no diabetes. Os tratamentos de ratos diabéticos com iogurte enriquecido com curcumina, licopeno ou bixina, isoladamente, melhoraram vários parâmetros alterados: fisiológicos (peso corporal e pesos de músculos esqueléticos e tecidos adiposos); bioquímicos (níveis plasmáticos de glicose, TAG e colesterol total foram diminuídos); as atividades de enzimas antioxidantes (SOD e CAT) foram aumentadas; biomarcadores de estresse oxidativo (MDA, PCO, LDL-ox) foram reduzidos, quando comparados com o grupo DIOG. Os tratamentos com os carotenoides isolados (DL e DB) aumentaram os níveis de colesterol-HDL. Os tratamentos com as misturas de curcumina com os carotenoides (DCB ou DCL) preservaram todos os efeitos benéficos dos tratamentos isolados. Além disso, a co-administração curcumina e carotenoides promoveu um efeito combinado na redução dos níveis de LDL-ox, bem como preservou os efeitos dos carotenoides no aumento dos níveis de HDL. Experimento II: No modelo de obesidade/resistência insulínica induzida com HF, foram utilizados camundongos machos (C57B1/6J), divididos aleatoriamente em 6 grupos, tratados durante 8 semanas: L, dieta low-fat e tratados com água; H, dieta high-fat e tratados com água; HIOG, dieta high-fat e tratados com iogurte; HC, dieta high-fat e tratados com 90 mg/kg de curcumina; HB, dieta high-fat e tratados com 5,5 mg/kg de bixina; HL, dieta high-fat e tratados com 45 mg/kg de licopeno. Todos os ativos naturais foram administrados em iogurte e em tratamentos isolados. Foram monitorados os parâmetros: i) fisiológicos: peso corporal, composição corporal via RMN (tecidos adiposos, massa magra e fluidos), massa de tecidos adiposos e musculares, ingestão alimentar, conteúdo energético das fezes; ii) biomarcadores plasmáticos: glicose [monitoramento da homeostase da glicose: teste de tolerância à insulina (ITT) e teste de tolerância a glicose oral (TTGO); glicemia de jejum (6 e 12 horas)], colesterol total, triacilglicerol (TAG), insulina (insulinemia de jejum e durante TTGO), níveis de MDA; iii) parâmetros bioquímicos teciduais: TAG, MDA, CAT, SOD, GSH-Px, e iv) biomarcadores inflamatórios teciduais: IL-6 e TNF-α hepáticos. Os animais do grupo H apresentaram piora nos parâmetros analisados, quando comparados aos animais dos grupos L, bem como com os demais grupos HIOG, HC, HB e HL, evidenciando assim a instalação do modelo de obesidade e resistência insulínica, acompanhado de estresse oxidativo e quadro inflamatório. Os camundongos tratados com iogurte enriquecido com curcumina (HC) ou licopeno (HL) apresentaram menor ganho de peso corporal e menor deposição de tecidos adiposos em relação ao grupo H, mostrando um importante papel destes ativos naturais na redução da adiposidade. Os animais dos grupos HIOG, HC, HL e HB também apresentaram menor acúmulo de TAG hepático. Os tratamentos com curcumina (HC), licopeno (HL) ou bixina (HB) também promoveram aumento nas atividades das enzimas antioxidantes (SOD, CAT e GSH-Px), e redução do biomarcador relacionado à lipoperoxidação (MDA). Os animais tratados com licopeno (HL) ou bixina (HB) apresentaram redução do biomarcador pró-inflamatório (TNF-α), quando comparados aos grupos HIOG e H. Além disso, o tratamento com licopeno (HL) também foi capaz de promover redução na resistência insulínica. A integração destes resultados nos permite concluir que os benefícios isolados ou combinados dos tratamentos com curcumina e carotenoides incorporados em iogurte substanciam as investigações e estimulam a necessidade em avançar nos estudos da utilização de misturas de ativos naturais, em especial aqueles que apresentam potencial antiaterogênico, anti-inflamatório e antioxidante, mitigando o risco cardiovascular.
The changes in lifestyle and in dietary structure of the current population lead to an excessive caloric intake and sedentary lifestyle, which can explain the epidemic proportions of obesity and diabetes mellitus (DM). DM is defined as a group of metabolic diseases characterized by hyperglycemia resulting from deficiency in the production and secretion of insulin by pancreatic beta cells and/or resistance to the insulin action in target tissues. The maintenance of hyperglycemia for long periods is a major factor in establishing the oxidative stress, which participates in the development of several impairments of diabetes, especially the microvascular and macrovascular complications. Many studies are demonstrating the effects of natural products on improving the hyperglycemia, oxidative stress, inflammatory status and the DM complications. Food matrices have been also explored as vehicle for natural actives, such as the yoghurt. In this context, we used two experimental models for assessing the effects of the treatments with curcumin, bixin or lycopene incorporated in yoghurt, isolated or in mixtures: i) type 1 DM induced by streptozotocin (STZ); ii) obesity/insulin resistance induced by high-fat (HF) diet. Experiment I: In DM model induced by STZ, male Wistar rats, normal (N) and diabetic (D, 40 mg/kg STZ, i.v.) animals, they were treated for 50 days, according to the following groups: NYOG and DYOG, treated with yoghurt; DINS, treated with insulin (4 U/day); DC, treated with 90 mg/kg curcumin; DB, treated with 5.5 mg/kg bixin; DCB, treated with 90 mg/kg curcumin and 5.5 mg/kg bixin; DL, treated with 45 mg/kg lycopene; DCL, treated with 90 mg/kg curcumin and 45 mg/kg lycopene. All natural compounds were administered with yoghurt. The following parameters were analyzed: i) physiological parameters: body weight, food and water intake, urinary volume, mass of epididymal and retroperitoneal white adipose tissues and soleus and extensor digitorum longus (EDL) skeletal muscles; ii) biochemical biomarkers in plasma, urine and tissues: glycemia, total cholesterol, HDL-cholesterol, triglycerides (TG), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), glycosuria, proteinuria and urinary urea, hepatic glycogen content; iii) plasma and tissue biomarkers of oxidative stress: thiobarbituric acid reactive substances (malondialdehyde, MDA), oxidized-LDL (ox-LDL), protein carbonyls (PCO), reduced glutathione (GSH) levels, and activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px). As expected, DYOG animals showed a worsening profile in all parameters studied during all the experimental period, while the treatment with insulin (DINS) was effective in improving all biomarkers changed by DM. The treatments of diabetic rats with curcumin, lycopene or bixin incorporated into yoghurt, alone, improved various parameters: physiological parameters (body weight and mass of adipose tissues and skeletal muscles), biochemical parameters (plasma levels of glucose, TG and total cholesterol were reduced); the activities of antioxidant enzymes (SOD and CAT) were increased; and oxidative stress biomarkers (MDA, PCO and oxLDL) were reduced, when compared with DYOG group. The treatments with isolated carotenoids (DL and DB) increased the HDL-cholesterol levels. The treatments with mixtures of curcumin and carotenoids (DCB or DCL) maintained all the beneficial effects of the individual treatments. In addition, the co-administration of curcumin and carotenoids promoted a combined effect in reducing the ox-LDL levels, and also maintained the carotenoids effects on the increase of HDL levels. Experiment II: In obesity/insulin resistance model, male mice (C57B1/6J) were divided randomly into 6 groups and they were treated for 8 weeks: L, low-fat diet and treated with water; H, high-fat diet and treated with water; HYOG, high-fat diet and treated with yoghurt; HC high-fat diet and treated with 90 mg/kg curcumin; HB high-fat diet and treated with 5.5 mg/kg bixin; HL, high-fat diet and treated with 45 mg/kg lycopene. All natural compounds were administered into yoghurt and in isolated treatments. The following parameters were monitored: i) physiological parameters: body weight, body composition via NMR (fat tissue, lean mass and fluids), mass of adipose tissues and skeletal muscles, food intake, energy content of feces; ii) plasma biomarkers: glucose [monitoring of glucose homeostasis: insulin tolerance test (ITT) and oral glucose tolerance test (OGTT); fasting glycemia (6-h and 12-h fasting)], total cholesterol, triglycerides (TG), insulin (fasting insulin and insulin during TTGO), MDA levels; iii) tissue biochemical parameters: TG, MDA, CAT, SOD, GSH-Px, and iv) inflammatory biomarkers: liver IL-6 and TNF-α. Animals of H group had a worsening profile in all the parameters analyzed, when compared with animals of L group, as well as with the other groups HYOG, HC, HB and HL, thus showing the installation of the obesity and insulin resistance. Mice treated with yoghurt enriched with curcumin (HC) or lycopene (HL) had low weight gain and minor deposition of adipose tissues, when compared with H group, showing an important role of these natural compounds in reducing the adiposity in HF-fed mice. In addition, HYOG, HC, HL and HB groups also had a minor TG accumulation in liver. The treatment with curcumin (HC), lycopene (HL) or bixin (HB) also promoted an increase in the activities of the antioxidant enzymes (SOD, CAT and GSH-Px), and a decrease in the lipoperoxidation biomarker (MDA). Animals treated with lycopene (HL) or bixin (HB) showed a decrease in the inflammatory biomarker (TNF-α), when compared with HYOG and H groups. In addition, the treatment with lycopene (HL) was also able to promote a decrease in the insulin resistance. The integration of these findings allow us to conclude that the isolated and the combined effects of the treatments of curcumin and carotenoids incorporated in yoghurt substantiate the investigations and stimulate the need to advance in the studies on the use of mixtures of natural compounds, mainly those having antiatherogenic, anti-inflammatory and antioxidant potentials, mitigating the cardiovascular risk.
The changes in lifestyle and in dietary structure of the current population lead to an excessive caloric intake and sedentary lifestyle, which can explain the epidemic proportions of obesity and diabetes mellitus (DM). DM is defined as a group of metabolic diseases characterized by hyperglycemia resulting from deficiency in the production and secretion of insulin by pancreatic beta cells and/or resistance to the insulin action in target tissues. The maintenance of hyperglycemia for long periods is a major factor in establishing the oxidative stress, which participates in the development of several impairments of diabetes, especially the microvascular and macrovascular complications. Many studies are demonstrating the effects of natural products on improving the hyperglycemia, oxidative stress, inflammatory status and the DM complications. Food matrices have been also explored as vehicle for natural actives, such as the yoghurt. In this context, we used two experimental models for assessing the effects of the treatments with curcumin, bixin or lycopene incorporated in yoghurt, isolated or in mixtures: i) type 1 DM induced by streptozotocin (STZ); ii) obesity/insulin resistance induced by high-fat (HF) diet. Experiment I: In DM model induced by STZ, male Wistar rats, normal (N) and diabetic (D, 40 mg/kg STZ, i.v.) animals, they were treated for 50 days, according to the following groups: NYOG and DYOG, treated with yoghurt; DINS, treated with insulin (4 U/day); DC, treated with 90 mg/kg curcumin; DB, treated with 5.5 mg/kg bixin; DCB, treated with 90 mg/kg curcumin and 5.5 mg/kg bixin; DL, treated with 45 mg/kg lycopene; DCL, treated with 90 mg/kg curcumin and 45 mg/kg lycopene. All natural compounds were administered with yoghurt. The following parameters were analyzed: i) physiological parameters: body weight, food and water intake, urinary volume, mass of epididymal and retroperitoneal white adipose tissues and soleus and extensor digitorum longus (EDL) skeletal muscles; ii) biochemical biomarkers in plasma, urine and tissues: glycemia, total cholesterol, HDL-cholesterol, triglycerides (TG), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), glycosuria, proteinuria and urinary urea, hepatic glycogen content; iii) plasma and tissue biomarkers of oxidative stress: thiobarbituric acid reactive substances (malondialdehyde, MDA), oxidized-LDL (ox-LDL), protein carbonyls (PCO), reduced glutathione (GSH) levels, and activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px). As expected, DYOG animals showed a worsening profile in all parameters studied during all the experimental period, while the treatment with insulin (DINS) was effective in improving all biomarkers changed by DM. The treatments of diabetic rats with curcumin, lycopene or bixin incorporated into yoghurt, alone, improved various parameters: physiological parameters (body weight and mass of adipose tissues and skeletal muscles), biochemical parameters (plasma levels of glucose, TG and total cholesterol were reduced); the activities of antioxidant enzymes (SOD and CAT) were increased; and oxidative stress biomarkers (MDA, PCO and oxLDL) were reduced, when compared with DYOG group. The treatments with isolated carotenoids (DL and DB) increased the HDL-cholesterol levels. The treatments with mixtures of curcumin and carotenoids (DCB or DCL) maintained all the beneficial effects of the individual treatments. In addition, the co-administration of curcumin and carotenoids promoted a combined effect in reducing the ox-LDL levels, and also maintained the carotenoids effects on the increase of HDL levels. Experiment II: In obesity/insulin resistance model, male mice (C57B1/6J) were divided randomly into 6 groups and they were treated for 8 weeks: L, low-fat diet and treated with water; H, high-fat diet and treated with water; HYOG, high-fat diet and treated with yoghurt; HC high-fat diet and treated with 90 mg/kg curcumin; HB high-fat diet and treated with 5.5 mg/kg bixin; HL, high-fat diet and treated with 45 mg/kg lycopene. All natural compounds were administered into yoghurt and in isolated treatments. The following parameters were monitored: i) physiological parameters: body weight, body composition via NMR (fat tissue, lean mass and fluids), mass of adipose tissues and skeletal muscles, food intake, energy content of feces; ii) plasma biomarkers: glucose [monitoring of glucose homeostasis: insulin tolerance test (ITT) and oral glucose tolerance test (OGTT); fasting glycemia (6-h and 12-h fasting)], total cholesterol, triglycerides (TG), insulin (fasting insulin and insulin during TTGO), MDA levels; iii) tissue biochemical parameters: TG, MDA, CAT, SOD, GSH-Px, and iv) inflammatory biomarkers: liver IL-6 and TNF-α. Animals of H group had a worsening profile in all the parameters analyzed, when compared with animals of L group, as well as with the other groups HYOG, HC, HB and HL, thus showing the installation of the obesity and insulin resistance. Mice treated with yoghurt enriched with curcumin (HC) or lycopene (HL) had low weight gain and minor deposition of adipose tissues, when compared with H group, showing an important role of these natural compounds in reducing the adiposity in HF-fed mice. In addition, HYOG, HC, HL and HB groups also had a minor TG accumulation in liver. The treatment with curcumin (HC), lycopene (HL) or bixin (HB) also promoted an increase in the activities of the antioxidant enzymes (SOD, CAT and GSH-Px), and a decrease in the lipoperoxidation biomarker (MDA). Animals treated with lycopene (HL) or bixin (HB) showed a decrease in the inflammatory biomarker (TNF-α), when compared with HYOG and H groups. In addition, the treatment with lycopene (HL) was also able to promote a decrease in the insulin resistance. The integration of these findings allow us to conclude that the isolated and the combined effects of the treatments of curcumin and carotenoids incorporated in yoghurt substantiate the investigations and stimulate the need to advance in the studies on the use of mixtures of natural compounds, mainly those having antiatherogenic, anti-inflammatory and antioxidant potentials, mitigating the cardiovascular risk.
Descrição
Palavras-chave
Diabetes mellitus, Obesidade, Estresse oxidativo, Curcumina, Licopeno, Bixina, Obesity, Oxidative stress, Curcumin, Lycopen, Bixin