Amentoflavone-Enriched Selaginella rossii Warb. Suppresses Body Weight and Hyperglycemia by Inhibiting Intestinal Lipid Absorption in Mice Fed a High-Fat Diet

Figures

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  Figure 1. High-performance liquid chromatography (HPLC) profiles of 95% EtOH extract (SRE) and EtOAc extract (SREA) of SR and amentoflavone (AMF). Chromatograms of HPLC for (A) SRE (5 mg/mL), and (B) SREA (5 mg/mL) were detected at 267 nm. (C) Chromatogram of HPLC, UV spectrum, and chemical structure of AMF.

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  Figure 2. Effects of SR extracts on the intestinal lipid absorption in the HFD-fed mice. (A) After SR extracts were administrated for seven days, a fat absorption test was performed after orogastric gavage of olive oil. Plasma TG concentration was measured at 0–4 h. (B) Area under the curve (AUC) of plasma TG concentration during the oral fat absorption test. (C) The food intake was measured during oral gavage of SR extracts for seven days. Fecal weight (D), fecal triglyceride (TG) (E), and free fatty acid (FFA) (F) contents were measured using the total fecal collected over 3 days. (G) The intestinal mRNA expressions were measured by quantitative real-time RT-PCR (qRT-PCR). Values are given as means ± SE (n = 7). * p < 0.05, ** p < 0.01 vs. the HFD group.

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  Figure 3. Effects of SR extracts on the body weight gain in the HFD-fed mice. (A) The body weights were monitored weekly during treatment of SR extracts for 10 weeks. (B,C) Weight gain and energy intake. (D) Images of the white adipose tissues (WATs) stained with H&E (×200 magnification, scale bar: 200 μm). (E) Adipocyte sizes of WATs. Values are given as means ± SE (n = 8). # p < 0.01 vs. the ND group; * p < 0.05, ** p < 0.01 vs. the HFD group. Retro AT, retroperitoneal AT.

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  Figure 4. Effects of SR extracts on glucose homeostasis and lipid profiles. (A) Fasting glucose levels were monitored during SR extracts treatment. (B) OGTT was performed by oral gavage of glucose. The blood glucose concentration was measured at 0–150 min. The AUCs of glucose concentration (C) and plasma insulin concentration (D) were measured during OGTT. The plasma insulin (E), blood glycated hemoglobin (HbA1c) (F), homeostasis model assessment of insulin resistance (HOMA-IR) (G), plasma TG (H), plasma aspartate transaminase (AST) (I), and alanine transaminase (ALT) (J) were analyzed at the end of the experiment. Values are given as means ± SE (n = 8). # p < 0.01 vs. the ND group; * p < 0.05, ** p < 0.01 vs. the HFD group.

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  Figure 5. Effects of SR extracts on intestinal lipid absorption and intestinal protein absorption. The mRNA expression levels related to lipid (A), carbohydrate (B), and protein (C) absorption were measured by real-time qRT-PCR. (D) The CD36 protein expression levels were measured using anti-CD36 and anti-GAPDH antibodies. Values are given as means ± SE (n = 8). # p < 0.01 vs. the ND group; * p < 0.05, ** p < 0.01 vs. the HFD group.

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  Figure 6. Effects of SR extracts on hepatic steatosis. (A,B) Images of liver were stained by H&E and Oil-Red O (× 200 magnification, scale bar: 200 μm). Hepatic TG (C) and TC (D) contents. (E–G) The hepatic mRNA expression was measured by qRT-PCR. Values are given as means ± SE (n = 8). # p < 0.01 vs. the ND group; * p < 0.05, ** p < 0.01 vs. the HFD group.

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  Figure 7. Effects of amentoflavone (AMF) on intestinal lipid absorption. (A) After AMF were administrated for seven days, a fat absorption test was performed after overnight fasting. Mice were given an orogastric gavage of olive oil. The plasma TG concentration was measured at 0, 2, and 4 h. The fecal TG (B) and FFA (C) contents were measured using the total fecal collected over 3 days. (D) The intestinal mRNA levels were measured by qRT-PCR. (E) The intestinal CD36 protein levels were measured using anti-CD36 and anti-GAPDH antibodies. Values are given as means ± SE (n = 6). * p < 0.05, ** p < 0.01 vs. the HFD group.