Nutrients, Vol. 14, Pages 5096: Timing of Medium-Chain Triglyceride Consumption Modulates Effects in Mice with Obesity Induced by a High-Fat High-Sucrose Diet

Figure 1. Effect of MCT intake on voluntary wheel-running activity in mice. (A) Hourly count/24 h of mouse wheel-running activity after 8 weeks of consuming HFHSD or M-HFSHD) (B) Daily wheel-running activity by mice fed with HFHSD or M-HFHSD. Data are shown as means ± SEM (n = 6). * p < 0.05 vs. M-HFHSD at the same time. HFHSD, high-fat high-sucrose diet; MCT, medium-chain triglyceride; M-HFHSD, high-fat high-sucrose diet containing medium-chain triglyceride; ZT, zeitgeber time.

Figure 1. Effect of MCT intake on voluntary wheel-running activity in mice. (A) Hourly count/24 h of mouse wheel-running activity after 8 weeks of consuming HFHSD or M-HFSHD) (B) Daily wheel-running activity by mice fed with HFHSD or M-HFHSD. Data are shown as means ± SEM (n = 6). * p < 0.05 vs. M-HFHSD at the same time. HFHSD, high-fat high-sucrose diet; MCT, medium-chain triglyceride; M-HFHSD, high-fat high-sucrose diet containing medium-chain triglyceride; ZT, zeitgeber time.

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Figure 2. Effects of MCT on body weight, food intake, and tissue weight. (A) Time course of changes in BW. (B) Average cumulative food intake by mice. Relative weights of the liver (C), Ga (D), eWAT (E), and sWAT (F) to BW. (G) Representative HE stained images of mouse eWAT. Scale bar = 200 μm. (H) Average area of adipocytes in eWAT from mice. Data were collected after feeding with HFHSD or M-HFSHD for 9 weeks. Data are shown as mean ± SEM (n = 8). * p < 0.05 and ** p < 0.01 vs. HFHSD. BW, body weight; Ga, gastrocnemius muscle; HFHSD, high-fat high-sucrose diet; MCT, medium-chain triglyceride; M-HFHSD, high-fat high-sucrose diet containing medium-chain triglyceride; eWAT, epididymal white adipose tissue; sWAT, inguinal subcutaneous white adipose tissue; HE, hematoxylin and eosin.

Figure 2. Effects of MCT on body weight, food intake, and tissue weight. (A) Time course of changes in BW. (B) Average cumulative food intake by mice. Relative weights of the liver (C), Ga (D), eWAT (E), and sWAT (F) to BW. (G) Representative HE stained images of mouse eWAT. Scale bar = 200 μm. (H) Average area of adipocytes in eWAT from mice. Data were collected after feeding with HFHSD or M-HFSHD for 9 weeks. Data are shown as mean ± SEM (n = 8). * p < 0.05 and ** p < 0.01 vs. HFHSD. BW, body weight; Ga, gastrocnemius muscle; HFHSD, high-fat high-sucrose diet; MCT, medium-chain triglyceride; M-HFHSD, high-fat high-sucrose diet containing medium-chain triglyceride; eWAT, epididymal white adipose tissue; sWAT, inguinal subcutaneous white adipose tissue; HE, hematoxylin and eosin.

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Figure 3. Serum hormone levels and metabolic parameters. Serum levels of insulin (A), leptin (B), glucose (C), β-hydroxybutyrate (D), NEFA (E), glycerol (F), triglyceride (G), and total cholesterol (H) at indicated times in mice fed with HFHSD or M-HFHSD for 9 weeks. Data are shown as mean ± SEM (n = 4). * p < 0.05 and ** p < 0.01 vs. HFHSD at same time. HFHSD, high-fat high-sucrose diet; MCT, medium-chain triglyceride; M-HFHSD, high-fat high-sucrose diet containing medium-chain triglyceride; NEFA, non-esterified fatty acid; ZT, zeitgeber time.

Figure 3. Serum hormone levels and metabolic parameters. Serum levels of insulin (A), leptin (B), glucose (C), β-hydroxybutyrate (D), NEFA (E), glycerol (F), triglyceride (G), and total cholesterol (H) at indicated times in mice fed with HFHSD or M-HFHSD for 9 weeks. Data are shown as mean ± SEM (n = 4). * p < 0.05 and ** p < 0.01 vs. HFHSD at same time. HFHSD, high-fat high-sucrose diet; MCT, medium-chain triglyceride; M-HFHSD, high-fat high-sucrose diet containing medium-chain triglyceride; NEFA, non-esterified fatty acid; ZT, zeitgeber time.

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Figure 4. Insulin and glucose tolerance tests and insulin signaling in peripheral tissues. Insulin tolerance tests at feeding period (A) or fasting period (B) in mice fed HFHSD or M-HFHSD for 8 weeks. Glucose tolerance test at feeding period (C) or fasting period (D) in mice fed HFHSD or M-HFSHD for 8 weeks. Protein expression levels of phosphorylated Akt on Ser473 or Thr307 in the liver (E), gastrocnemius muscle (F), and epididymal white adipose tissue (G) extracts from mice. Data are mean ± SEM (n = 4). * p < 0.05 vs. HFHSD at same time. # p < 0.05 and ## p < 0.01 vs. same diet at ZT8. HFHSD, high-fat high-sucrose diet; MCT, medium-chain triglyceride; M-HFHSD, high-fat high-sucrose diet containing medium-chain triglyceride; ZT, zeitgeber time.

Figure 4. Insulin and glucose tolerance tests and insulin signaling in peripheral tissues. Insulin tolerance tests at feeding period (A) or fasting period (B) in mice fed HFHSD or M-HFHSD for 8 weeks. Glucose tolerance test at feeding period (C) or fasting period (D) in mice fed HFHSD or M-HFSHD for 8 weeks. Protein expression levels of phosphorylated Akt on Ser473 or Thr307 in the liver (E), gastrocnemius muscle (F), and epididymal white adipose tissue (G) extracts from mice. Data are mean ± SEM (n = 4). * p < 0.05 vs. HFHSD at same time. # p < 0.05 and ## p < 0.01 vs. same diet at ZT8. HFHSD, high-fat high-sucrose diet; MCT, medium-chain triglyceride; M-HFHSD, high-fat high-sucrose diet containing medium-chain triglyceride; ZT, zeitgeber time.

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Figure 5. Expression of lipogenic genes and proteins in the liver. Expression levels of Fasn (A), Acc1 (B), Srebp1c (C), and Pparg1 (D) mRNA at indicated times in the liver of mice. (E) Protein expression levels of FAS, ACC, PPARγ1 and GAPDH in liver extracts from mice. (FH) Ratios of FAS-, ACC-, and PPARγ1-to-GAPDH. Data are mean ± SEM (n = 4). * p < 0.05 and ** p < 0.01 vs. HFHSD at same time. # p < 0.05 vs. same diet at ZT8. HFHSD, high-fat high-sucrose diet; MCT, medium-chain triglyceride; M-HFHSD, high-fat high-sucrose diet containing medium-chain triglyceride; ZT, zeitgeber time.

Figure 5. Expression of lipogenic genes and proteins in the liver. Expression levels of Fasn (A), Acc1 (B), Srebp1c (C), and Pparg1 (D) mRNA at indicated times in the liver of mice. (E) Protein expression levels of FAS, ACC, PPARγ1 and GAPDH in liver extracts from mice. (FH) Ratios of FAS-, ACC-, and PPARγ1-to-GAPDH. Data are mean ± SEM (n = 4). * p < 0.05 and ** p < 0.01 vs. HFHSD at same time. # p < 0.05 vs. same diet at ZT8. HFHSD, high-fat high-sucrose diet; MCT, medium-chain triglyceride; M-HFHSD, high-fat high-sucrose diet containing medium-chain triglyceride; ZT, zeitgeber time.

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Figure 6. Expression of lipogenic genes and proteins in epididymal WAT. Expression levels of Fasn (A), Acc1 (B), Srebp1c (C), and Pparγ2 (D) mRNA at indicated times in epididymal WATs of mice. (E) Protein expression levels of FAS, ACC, PPARγ1/2, and GAPDH in eWAT extracts from mice. (FH) Ratios of FAS-, ACC-, and PPARγ2-to-GAPDH. Data are mean ± SEM (n = 4). * p < 0.05 and ** p < 0.01 vs. HFHSD at same time. # p < 0.05 and ## p < 0.01 vs. same diet at ZT8. WAT, white adipose tissue; eWAT, epididymal white adipose tissue; HFHSD, high-fat high-sucrose diet; MCT, medium-chain triglyceride; M-HFHSD, high-fat high-sucrose diet containing medium-chain triglyceride; ZT, zeitgeber time.

Figure 6. Expression of lipogenic genes and proteins in epididymal WAT. Expression levels of Fasn (A), Acc1 (B), Srebp1c (C), and Pparγ2 (D) mRNA at indicated times in epididymal WATs of mice. (E) Protein expression levels of FAS, ACC, PPARγ1/2, and GAPDH in eWAT extracts from mice. (FH) Ratios of FAS-, ACC-, and PPARγ2-to-GAPDH. Data are mean ± SEM (n = 4). * p < 0.05 and ** p < 0.01 vs. HFHSD at same time. # p < 0.05 and ## p < 0.01 vs. same diet at ZT8. WAT, white adipose tissue; eWAT, epididymal white adipose tissue; HFHSD, high-fat high-sucrose diet; MCT, medium-chain triglyceride; M-HFHSD, high-fat high-sucrose diet containing medium-chain triglyceride; ZT, zeitgeber time.

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Figure 7. Expression of lipolytic genes and proteins and lipolysis-related signaling in epididymal white adipose tissue. Expression levels of Atgl (A) and Hsl (B) mRNA at indicated times in epididymal white adipose tissues of mice. (C) Protein expression of ATGL, phosphorylated HSL on Ser563, phosphorylated HSL on Ser660, total HSL and GAPDH in epididymal white adipose tissue extracts from mice. (DF) Ratios of ATGL-to-GAPDH and phosphorylated protein-to-total protein. (G) Protein expression of phosphorylated PKA substrate, phosphorylated ERK, total ERK, and GAPDH in epididymal white adipose tissue extracts from mice. (H,I) Ratios of phosphorylated PKA substrate-to-GAPDH and phosphorylated ERK-to-total ERK. Data are mean ± SEM (n = 4). * p < 0.05 and ** p < 0.01 vs. HFHSD at same time. # p < 0.05 and ## p < 0.01 vs. same diet at ZT8. HFHSD, high-fat high-sucrose diet; MCT, medium-chain triglyceride; M-HFHSD, high-fat high-sucrose diet containing medium-chain triglyceride; ZT, zeitgeber time.

Figure 7. Expression of lipolytic genes and proteins and lipolysis-related signaling in epididymal white adipose tissue. Expression levels of Atgl (A) and Hsl (B) mRNA at indicated times in epididymal white adipose tissues of mice. (C) Protein expression of ATGL, phosphorylated HSL on Ser563, phosphorylated HSL on Ser660, total HSL and GAPDH in epididymal white adipose tissue extracts from mice. (DF) Ratios of ATGL-to-GAPDH and phosphorylated protein-to-total protein. (G) Protein expression of phosphorylated PKA substrate, phosphorylated ERK, total ERK, and GAPDH in epididymal white adipose tissue extracts from mice. (H,I) Ratios of phosphorylated PKA substrate-to-GAPDH and phosphorylated ERK-to-total ERK. Data are mean ± SEM (n = 4). * p < 0.05 and ** p < 0.01 vs. HFHSD at same time. # p < 0.05 and ## p < 0.01 vs. same diet at ZT8. HFHSD, high-fat high-sucrose diet; MCT, medium-chain triglyceride; M-HFHSD, high-fat high-sucrose diet containing medium-chain triglyceride; ZT, zeitgeber time.

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Table 1. Composition of the diets.

Table 1. Composition of the diets.

Ingredients (g/100 g)HFHSDM-HFHSDCasein2525Cystine0.3750.375Cornstarch14.86914.869Sucrose2020Beef tallow147Lard148MCT oil013Soybean oil22Cellulose55Mineral mixture, AIN-933.53.5Vitamin mixture, AIN-9311Choline bitartrate0.250.25Tert-butyl hydroquinone0.0060.006Fat (% of energy)53.953.9Total energy (MJ/100 g diet)1.991.99

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