A Novel Identified Peptide Hormone "Metabolitin" Attenuates Lipid Absorption in the Small Intestine of Diabetic Mice with Nonalcoholic Fatty Liver Disease by Regulating Neurotensin and AMPK Signaling Pathway.

Aim: The purpose of this study was to explore the effect of a novel identified peptide hormone "metabolitin" on lipid absorption in the small intestine of mice with type 2 diabetes mellitus (T2DM) and nonalcoholic fatty liver disease (NAFLD) and potential mechanisms.

Methods: T2DM was induced in mice by 4-6 weeks of high-fat diets followed by intraperitoneal injection of 35 mg/kg STZ. NAFLD was induced in diabetic mice by a month of high-fat diets. Oral administration of 4 pmol/g or 12 pmol/g metabolitin every two days was performed during one-month high-fat diets. Triglyceride (TG) and total cholesterol (TC) detection and Oil Red O staining were performed to evaluate lipid absorption. The neurotensin (NT) levels in the intestinal tissues and serum were determined by ELISA. Lipogenesis- and lipolysis-related proteins, AMP-activated protein kinase (AMPK), and p-AMPK were examined by Western blot analysis.

Results: It was found that glucose tolerance test (GTT), insulin tolerance test (ITT), TG, and TC indicated lower levels in the serum of NAFLD/T2DM mice receiving 4 pmol/g and 12 pmol/g metabolitin compared to the mice receiving normal saline ( P < 0.05). No significant difference was noted in the TC level of the feces among mice with different diets ( P > 0.05), but compared to NAFLD/T2DM mice with normal saline, the mice administrated with 4 pmol/g and 12 pmol/g metabolitin revealed much higher TG levels in the feces ( P < 0.05). The results of Oil Red O staining revealed that the intestinal epithelial cells of NAFLD/T2DM mice receiving 12 pmol/g metabolitin indicated resistance to lipid absorption and the area of staining was smaller than that of NAFLD/T2DM mice with normal saline ( P < 0.05). The NAFLD/T2DM mice receiving 4 pmol/g and 12 pmol/g metabolitin showed a higher extent of p-AMPK concomitant with lower levels of NT in the serum and small intestine than the mice with normal saline ( P < 0.05). Western blot analysis also suggested that NAFLD/T2DM mice receiving 4 pmol/g and 12 pmol/g metabolitin revealed lower expressions in fatty acid synthase (FAS), acetyl-CoA carboxylase (ACC), stearoyl-CoA desaturase-1 (SCD-1), and sterol regulatory element-binding transcription factor-1 (SREBP1) proteins and higher expressions in carnitine palmitoyltransferase 1 (CPT1), peroxisome proliferator-activated receptor alpha (PPAR α ), and fatty acid translocase (CD36) proteins than NAFLD/T2DM mice with normal saline ( P < 0.05).

Conclusion: According to the data we observed, oral administration of metabolitin could attenuate lipid absorption in the small intestine of NAFLD/T2DM mice, which may be a novel therapeutic approach for NAFLD/T2DM.