Yunpi Heluo decoction attenuates insulin resistance by regulating SIRT1-FoxO1 autophagy pathway in skeletal muscle of Zucker diabetic fatty rats

Abstract

Ethnopharmacological relevance: Diabetes is a serious chronic metabolic disorder, and type 2 diabetes mellitus (T2DM) accounts for more than 90% of all diabetes cases. Insulin resistance (IR) is an early symptom, typical feature and main pathogenesis of T2DM due to the combined effects of genetic and environmental factors. Current evidence shows that IR is mainly caused by nutrient overload, systemic fatty acid excess, fatty tissue inflammation, endoplasmic reticulum stress, oxidative stress and abnormal autophagy. Autophagy plays an important role in the development of IR and decreased autophagy activity can cause IR through various ways.

Aim of the study: Yunpiheluo (YPHL) decoction is a Chinese herbal formula with unique advantages for the treatment of T2DM. The aim of the present study was to investigate the regulatory mechanism of YPHL on the autophagy pathway in the skeletal muscle of IR Zucker diabetic fatty (ZDF) rats.

Methods: T2DM ZDF rats were treated with YPHL or transfected with SIRT1 adeno-associated virus. Serum total cholesterol (TC), triglyceride (TG), high-density lipoprotein (HDL), low-density lipoprotein (LDL), insulin resistance index (IRI) and skeletal muscle TG levels were detected in a T2DM ZDF rat model. The skeletal muscle morphology was observed by histological analysis and Oil Red O Staining. Autophagosomes were observed by transmission electron microscopy (TEM). The skeletal muscle morphology and fat deposition were observed by histological examination and Oil Red O Staining. A rat skeletal muscle IR cell model was established and transfected with SIRT1 overexpression plasmids. Cell apoptosis was observed by DAPI staining. SIRT1 levels in skeletal muscle tissues and cells were detected by qRT-PCR. The protein expressions of SIRT1, FOXo1, LC3B and P62 were detected by Western blotting.

Results: Large numbers of lipid droplets and swollen mitochondria were observed in the skeletal muscle in both model group and negative control (NC) group receiving blank plasmid. Autophagosomes were seen in the skeletal muscle of YPHL and SIRT1 groups, with no significant structural abnormality. In addition, the protein expression of LC3B was decreased and the protein expression of p62 was increased significantly in the model group as compared with the NC group. After intervention with YPHL and SIRT1 overexpression, the protein expression of LC3B was significantly increased and p62 was significantly decreased. However, there was no significant difference in cell apoptosis between the two groups.

Conclusion: The SIRT1-FoxO1 autophagy pathway may play a significant role in the pathogenesis of IR. YPHL could increase the autophagy level by regulating the SIRT1-FoxO1 signaling pathway in the skeletal muscle and improving the lipid metabolism, thereby attenuating IR.

Keywords: Autophagy; Autophagy pathway; Insulin resistance; SIRT1-FoxO1; Skeletal muscle; Yunpiheluo decoction.