Icariin improves brain function decline in aging rats by enhancing neuronal autophagy through the AMPK/mTOR/ULK1 pathway

Pharm Biol. 2021 Dec;59(1):183-191. doi: 10.1080/13880209.2021.1878238.

Abstract

Context: Icariin (ICA) is the main active ingredient of Epimedium brevicornu Maxim (Berberidaceae), which is used in the immune, reproductive, neuroendocrine systems, and anti-aging.

Objective: To evaluate the effect of ICA on natural aging rat.

Materials and methods: 16-month-old Sprague-Dawley (SD) rats were randomly divided into aging, low and high-dose ICA groups (n = 8); 6-month-old rats were taken as the adult control (n = 8). Rats were fed regular feed (aging and adult control) or feed containing ICA (ICA 2 and 6 mg/kg group) for 4 months. HE and Nissl staining were used to assess pathological changes. Western blot was used to test the expression of autophagy (LC3B, p62, Atg5, Beclin1) and p-AMPK, p-mTOR and p-ULK1 (ser 757). Immunofluorescence was used to detect the co-localization of LC3 and neurons.

Results: ICA improved neuronal degeneration associated with aging and increased the staining of Nissl bodies. Western blot showed that ICA up-regulated autophagy-related proteins LC3B (595%), Beclin1 (73.5%), p-AMPK (464%) protein (p < 0.05 vs. 20 M) in the cortex and hippocampus of aging rats, down-regulated the expression of p62 (56.9%), p-mTOR (53%) and p-ULK1 (ser 757) (65.4%) protein (p < 0.05 vs. 20 M). Immunofluorescence showed that the fluorescence intensity of LC3 decreased in the aging rat brain, but increased and mainly co-localized with neurons after ICA intervention.

Conclusions: Further research needs to verify the expression changes of AMPK/mTOR/ULK1 and the improvement effect of ICA in elderly. These results will further accelerate the applications of ICA and the treatment for senescence.

Keywords: Epimedium brevicornu Maxim; Senescence; macroautophagy; neurodegenerative diseases.

Publication types

  • Comparative Study

MeSH terms

  • AMP-Activated Protein Kinases / metabolism
  • Aging / physiology
  • Animals
  • Autophagy / drug effects*
  • Autophagy-Related Protein-1 Homolog / metabolism
  • Brain / drug effects*
  • Brain / pathology
  • Dose-Response Relationship, Drug
  • Epimedium / chemistry
  • Flavonoids / administration & dosage
  • Flavonoids / isolation & purification
  • Flavonoids / pharmacology*
  • Neurons / drug effects*
  • Neurons / pathology
  • Rats
  • Rats, Sprague-Dawley
  • TOR Serine-Threonine Kinases / metabolism

Substances

  • Flavonoids
  • mTOR protein, rat
  • Autophagy-Related Protein-1 Homolog
  • TOR Serine-Threonine Kinases
  • ULK1 protein, rat
  • AMP-Activated Protein Kinases
  • icariin

Grants and funding

This work was supported by National Natural Science Foundation of China [Grant Number: 81673778, 1212202].