Nutr Res Pract.  2013 Aug;7(4):294-301.

Dietary carnosic acid suppresses hepatic steatosis formation via regulation of hepatic fatty acid metabolism in high-fat diet-fed mice

• Park MY
• Mun ST

Abstract

In this study, we examined the hepatic anti-steatosis activity of carnosic acid (CA), a phenolic compound of rosemary (Rosmarinus officinalis) leaves, as well as its possible mechanism of action, in a high-fat diet (HFD)-fed mice model. Mice were fed a HFD, or a HFD supplemented with 0.01% (w/w) CA or 0.02% (w/w) CA, for a period of 12 weeks, after which changes in body weight, blood lipid profiles, and fatty acid mechanism markers were evaluated. The 0.02% (w/w) CA diet resulted in a marked decline in steatosis grade, as well as in homeostasis model assessment of insulin resistance (HOMA-IR) index values, intraperitoneal glucose tolerance test (IGTT) results, body weight gain, liver weight, and blood lipid levels (P < 0.05). The expression level of hepatic lipogenic genes, such as sterol regulating element binding protein-1c (SREBP-1c), liver-fatty acid binding protein (L-FABP), stearoyl-CoA desaturase 1 (SCD1), and fatty acid synthase (FAS), was significantly lower in mice fed 0.01% (w/w) CA and 0.02% (w/w) CA diets than that in the HFD group; on the other hand, the expression level of beta-oxidation-related genes, such as peroxisome proliferator-activated receptor alpha (PPAR-alpha), carnitine palmitoyltransferase 1 (CPT-1), and acyl-CoA oxidase (ACO), was higher in mice fed a 0.02% (w/w) CA diet, than that in the HFD group (P < 0.05). In addition, the hepatic content of palmitic acid (C16:0), palmitoleic acid (C16:1), and oleic acid (C18:1) was significantly lower in mice fed the 0.02% (w/w) CA diet than that in the HFD group (P < 0.05). These results suggest that orally administered CA suppressed HFD-induced hepatic steatosis and fatty liver-related metabolic disorders through decrease of de novo lipogenesis and fatty acid elongation and increase of fatty acid beta-oxidation in mice.

Keyword

Carnosic acid; high-fat diet; hepatic steatosis; lipogenesis; fatty acid oxidation

MeSH Terms

Acyl Coenzyme A
Acyl-CoA Oxidase
Animals
Body Weight
Carnitine O-Palmitoyltransferase
Carrier Proteins
Diet
Diet, High-Fat
Diterpenes, Abietane
Fatty Acid Synthetase Complex
Fatty Acids, Monounsaturated
Glucose Tolerance Test
Hand
Homeostasis
Insulin Resistance
Lipogenesis
Liver
Mice
Oleic Acid
Palmitic Acid
Phenol
Plant Extracts
PPAR alpha
Stearoyl-CoA Desaturase
Acyl Coenzyme A
Acyl-CoA Oxidase
Carnitine O-Palmitoyltransferase
Carrier Proteins
Diterpenes, Abietane
Fatty Acid Synthetase Complex
Fatty Acids, Monounsaturated
Oleic Acid
PPAR alpha
Palmitic Acid
Phenol
Plant Extracts
Stearoyl-CoA Desaturase

Figure

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  Fig. 1 Effect of carnosic acid supplementation on high-fat diet-induced body weight gain and tissue (liver and white adipose tissue) weight. Mice were fed either a normal diet (ND) or a high-fat diet (HFD), alone or supplemented with 0.01% (w/w) carnosic acid (CA) or 0.02% (w/w) CA (HFD + CA) for 12 weeks. (A) Body weight gain, (B) food intake, (C) liver weight, and (D) white adipose tissue (WAT) weight were measured for all four groups of mice. Values are expressed as mean ± SD (n = 10). Bars with different letters differ significantly (P < 0.05) from one another.

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  Fig. 2 Effect of carnosic acid supplementation on hepatic steatosis score and adipocyte size in white adipose tissue. (A) Representative histological sections of liver tissue, (B) steatosis grade score, (C) representative histological sections of white adipose tissue (WAT) tissues, and (D) relative adipose size in WAT tissues were evaluated for all four groups of mice. Mice were fed a normal diet (ND) or a high-fat diet (HFD), alone or supplemented with 0.01% (w/w) carnosic acid (CA) or 0.02% (w/w) CA (HFD + CA) for 12 weeks. Liver tissue sections obtained at the end of the 12 weeks were stained with hematoxylin and eosin (×400).

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  Fig. 3 Effect of carnosic acid supplementation on blood glucose and insulin levels in high-fat diet-fed mice. Mice were fed a normal diet (ND) or a high-fat diet (HFD), alone or supplemented with 0.01% (w/w) carnosic acid (CA) or 0.02% (w/w) CA (HFD + CA) for 12 weeks. (A) Glucose tolerance test was performed through intraperitoneal injection of glucose (2 g/kg body weight) into mice, and blood glucose levels were measured 0, 30, 60, and 120 min later, (B) percentage of area under the curve (%AUC), (C) serum insulin levels, and (D) insulin resistance index were evaluated for all four groups. Values are expressed as mean ± SD (n = 10). Bars with different letters differ significantly (P < 0.05) from one another.

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  Fig. 4 Effect of carnosic acid supplementation on blood lipid levels in high-fat diet-fed mice. Mice were fed a normal diet (ND) or a high-fat diet (HFD), alone or supplemented with 0.01% (w/w) carnosic acid (CA) or 0.02% (w/w) CA (HFD + CA) for 12 weeks. (A) Free fatty acid (FFA), (B) triglyceride (TG), (C) total-cholesterol (T-CHO), (D) low-density lipoprotein (LDL), and (E) high-density lipoprotein (HDL) levels, as well as (F) the ratio of high-density lipoprotein to total-cholesterol (HDL/T-CHO) in serum were calculated for all four groups. Values are expressed as mean ± SD (n = 10). Bars with different letters differ significantly (P < 0.05) from one another.

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  Fig. 5 Effects of carnosic acid supplementation on hepatic mRNA expression of genes related to fatty acid synthesis and β-oxidation in high-fat diet-fed mice. Mice were fed a normal diet (ND) or a high-fat diet (HFD), alone or supplemented with 0.01% (w/w) carnosic acid (CA) or 0.02% (w/w) CA (HFD + CA) for 12 weeks. Consequently, the following four groups of mice (n = 10, each) were examined: control ND group (white), HFD group (black), HFD supplemented with 0.01% (w/w) CA group (dark grey), and HFD supplemented with 0.02% (w/w) CA group (light grey). Analysis of hepatic expression of various genes was performed using real-time PCR, and expression levels were normalized to β-actin. Expression levels of (A) lipogenic genes (Srebp-1c, sterol regulatory element-binding protein-1c; L-fabp, liver-fatty acid binding protein; Scd1, stearoyl-CoA desaturase 1; Fas, fatty acid synthase), and (B) β-oxidation-related genes (Ppar-α, peroxisome proliferator-activated receptor-α; Cpt1, carnitine palmitoyltransferase 1; Aco, acyl CoA oxidase) are represented here. Values are expressed as mean ± SD (n = 10). Bars with different letters differ significantly (P < 0.05) from one another.

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