Ramipril significantly attenuates the development of non-alcoholic steatohepatitis in hyperlipidaemic rabbits.

BACKGROUND: Non-alcoholic fatty liver disease (NAFLD) is considered as the most frequent cause of chronic hepatic disease in adults. It is strictly correlated with insulin resistance, frequently associated with components of metabolic syndrome, and, similarly to the latter, it has been correlated with high risk of developing type 2 diabetes and cardiovascular diseases. Systemic arterial hypertension has been suggested to be associated with NAFLD in approximately 40% of the cases, and NAFLD has been independently associated with an increased risk of arterial hypertension in observational studies. Therefore, we can infer that treating arterial hypertension in NAFLD carriers will be often necessary and that the potential beneficial effects of the antihypertensive might, in this context, influence the choice of the respective drug. The renin-angiotensin system has been correlated to the whole basic physiopathogenic mechanism of NAFLD in experimental models. Based on these findings, we conducted this study to evaluate the effects of the ACE-inhibitor ramipril, used preventively, in NAFLD induced in rabbits fed hyperlipidaemic diet. METHODS: Twenty-nine rabbits were divided into three groups (normal, placebo, and ramipril). The placebo and ramipril groups were fed a ration containing 0.925% cholesterol. The groups were orally administered 0.35 mg/kg/day of ramipril, and an equivalent volume of vehicle was administered to the placebo group. At the end of the 8th week, all rabbits underwent segmental hepatic resection and were euthanized. Blood samples were collected to determine glucose, insulin, creatinine, total cholesterol, triglycerides, HDL-C, and aminotransferase levels at baseline and euthanasia. Haematoxylin and eosin and Gomori trichrome-stained slides were analysed based on the histological scoring system for NAFLD. Sudan III-stained slides were analysed by morphometry and immunostained based on the Allred scoring system. RESULTS: When compared with placebo, ramipril significantly diminished the development of steatosis (P=0.032), lobular inflammation (P=0.006), hepatocellular ballooning (P=0.023), and fibrosis (P=0.02). Based on the NAFLD activity score (NAS), ramipril significantly reduced the development of non-alcoholic steatohepatitis (NASH) (P=0.003). CONCLUSIONS: The preventive use of ramipril in rabbits fed hyperlipidaemic diet, attenuates the development of the whole NAFLD histopathological spectrum and based on NAS, ramipril significantly reduced the development of NASH.

Effect of olmesartan on leukocyte recruitment in choroid-sclera complex in hypercholesterolemia model.

PURPOSE: Demonstrate that the blockade of angiotensin II AT-1 receptors, through the systemic administration of olmesartan, can reduce the MCP-1 expression and the resulting macrophage accumulation in the choroid and sclera of hypercholesterolemic rabbits. METHODS: Thirty-two New Zealand rabbits were divided into 3 groups: group I (GI) was fed a standard rabbit diet; group II (GII) was fed a hypercholesterolemic diet; and group III (GIII) was fed a hypercholesterolemic diet plus olmesartan. Serum levels of total cholesterol, triglyceride, HDL cholesterol, and blood glucose were determined in fasting rabbits at the beginning of the experiment and on the day of euthanasia. The choroid and sclera were submitted to morphometric analysis as well as immunohistochemical analysis with MCP-1 and RAM-11 (macrophage marker) antibodies. RESULTS: No abnormality was detected in GI. Group II and III had significant increases in choroid-sclera complex thicknesses when compared with group I (P<0.001). GII showed a significant increase in immunoreactivity for MCP-1 in relation to GI (P=0.001) and GIII (P=0.004). GII showed a significant increase in immunoreactivity for RAM-11 of the choroid-sclera complex in relation to GI (P<0.001) and GIII (P=0.034). A significant increase in immunoreactivity for RAM-11 was observed in GIII in relation to GI (P=0.008). CONCLUSION: Olmesartan reduced the MCP-1 expression and the resultant macrophage accumulation in the choroid-sclera complex of hypercholesterolemic rabbits.

[Increase of choroidal and scleral MCP-1 expression in hypercholesterolemia experimental model]. / Aumento da expressão do MCP-1 coroidal e escleral em modelo experimental de hipercolesterolemia.

PURPOSE: The aim of this study is to experientially demonstrate that a cholesterol-enriched diet induces an increase in the MCP-1 expression in the choroid and sclera. METHOD: New Zealand rabbits were divided into two groups: GN (normal diet group) of 8 rabbits (8 eyes) was fed a standard rabbit diet for 4 weeks; GH (hypercholesterolemic group) of 13 rabbits (13 eyes) was fed a 1% cholesterol enriched diet for 8 weeks. Total serum cholesterol, triglyceride, HDL cholesterol and fasting blood glucose exams were performed at the start of the experiment and at the euthanasia time. After GH 8th week and GN 4th week animals were euthanized and their eyes underwent immunohistochemical analysis with the anti-MCP-1 antibody. RESULTS: The diet has induced a significant increase in GH total cholesterol and triglyceride levels when compared with NG (p<0.001). There was a significant increase in the MCP-1 expression in GH choroid and sclera in relation to GN (p<0.001). CONCLUSION: This study has revealed that the hypercholesterolemic diet in rabbits induces an increase in the MCP-1 expression in the choroid and sclera.

Aumento da expressão do MCP-1 coroidal e escleral em modelo experimental de hipercolesterolemia / Increase of choroidal and scleral MCP-1 expression in hypercholesterolemia experimental model

OBJETIVO: O objetivo deste trabalho é demonstrar experimentalmente que a dieta rica em colesterol provoca aumento da expressão da MCP-1 na coroide e esclera. MÉTODO: Coelhos New Zealand foram organizados em dois grupos: GN (grupo dieta normal), composto por 8 coelhos (8 olhos), recebeu ração padrão para coelhos, durante 4 semanas; GH (grupo hipercolesterolêmico), composto por 13 coelhos (13 olhos), recebeu dieta rica em colesterol a 1% por 8 semanas. Foi realizada a dosagem sérica de colesterol total, triglicerídeos, HDL colesterol, glicemia de jejum no início do experimento e no momento da eutanásia. Ao final da 8ª semana para o GH e 4ª semana para o GN foi realizada a eutanásia dos animais e os olhos foram submetidos à análise imuno-histoquímica com o anticorpo anti-MCP-1. RESULTADOS: A dieta provocou significativo aumento do colesterol total e triglicerídeos do GH em relação ao GN (p<0,001). Houve significativo aumento da expressão da MCP-1 na coroide e esclera dos animais do GH em relação ao GN (p<0,001). CONCLUSÃO: Este estudo demonstrou que a dieta hipercolesterolêmica em coelhos induz ao aumento da expressão do MCP-1 na coroide e esclera.

PURPOSE: The aim of this study is to experientially demonstrate that a cholesterol-enriched diet induces an increase in the MCP-1 expression in the choroid and sclera. METHOD: New Zealand rabbits were divided into two groups: GN (normal diet group) of 8 rabbits (8 eyes) was fed a standard rabbit diet for 4 weeks; GH (hypercholesterolemic group) of 13 rabbits (13 eyes) was fed a 1% cholesterol enriched diet for 8 weeks. Total serum cholesterol, triglyceride, HDL cholesterol and fasting blood glucose exams were performed at the start of the experiment and at the euthanasia time. After GH 8th week and GN 4th week animals were euthanized and their eyes underwent immunohistochemical analysis with the anti-MCP-1 antibody. RESULTS: The diet has induced a significant increase in GH total cholesterol and triglyceride levels when compared with NG (p<0.001). There was a significant increase in the MCP-1 expression in GH choroid and sclera in relation to GN (p<0.001). CONCLUSION: This study has revealed that the hypercholesterolemic diet in rabbits induces an increase in the MCP-1 expression in the choroid and sclera.

Olmesartan severely weakened the development of NASH in an animal model of hypercholesterolemia.

BACKGROUND: Non-alcoholic fatty liver disease (NAFLD) is characterized by a broad spectrum of liver damage. In a rat model of non-alcoholic steatohepatitis (NASH), olmesartan attenuated steatosis and fibrosis. OBJECTIVE: To assess the potential preventive action of olmesartan, an angiotensin II type 1 receptor blocker, on NAFLD in hypercholesterolemic rabbits. METHODS: Thirty-four white adult male rabbits were selected. The animals were divided into three groups: group I (GI), control group, 13 rabbits; group II (GII), olmesartan group, 12 rabbits; and group III (GIII), normal group, 9 rabbits. The animals from GI and GII were fed with a specific diet plus 1% cholesterol. Animals from GIII were fed only with a specific diet. The GII animals were treated with olmesartan. RESULTS: Steatosis was present in all animals from GI and GII; no steatosis was observed in animals from GIII. When GI and GII where compared, the steatosis had higher scores in GI (p < 0.013). Perisinusoidal and periportal fibrosis occurred in 46.2% of the animals from GI. There was no fibrosis in GII or GIII. Lobular inflammation occurred in 84.6% of the animals from GI. Animals from GII and GIII had no inflammation. The NAFLD activity score was higher in animals from GI when compared to animals from GII and GIII (p < 0.001 for both groups); the NAFLD score was significantly higher in animals from GII when compared to animals from GIII (p < 0.001). CONCLUSIONS: In hypercholesterolemic rabbits, olmesartan significantly attenuated hepatic steatosis and prevented the development of lobular inflammation and liver fibrosis. Based on the NAFLD activity score, olmesartan significantly weakened the development of NASH in rabbits fed a high cholesterol diet.