Andrographolide protects mouse astrocytes against hypoxia injury by promoting autophagy and S100B expression

Braz J Med Biol Res. 2018;51(6):e7061. doi: 10.1590/1414-431x20177061. Epub 2018 Apr 19.

Abstract

Andrographolide (ANDRO) has been studied for its immunomodulation, anti-inflammatory, and neuroprotection effects. Because brain hypoxia is the most common factor of secondary brain injury after traumatic brain injury, we studied the role and possible mechanism of ANDRO in this process using hypoxia-injured astrocytes. Mouse cortical astrocytes C8-D1A (astrocyte type I clone from C57/BL6 strains) were subjected to 3 and 21% of O2 for various times (0-12 h) to establish an astrocyte hypoxia injury model in vitro. After hypoxia and ANDRO administration, the changes in cell viability and apoptosis were assessed using CCK-8 and flow cytometry. Expression changes in apoptosis-related proteins, autophagy-related proteins, main factors of JNK pathway, ATG5, and S100B were determined by western blot. Hypoxia remarkably damaged C8-D1A cells evidenced by reduction of cell viability and induction of apoptosis. Hypoxia also induced autophagy and overproduction of S100B. ANDRO reduced cell apoptosis and promoted cell autophagy and S100B expression. After ANDRO administration, autophagy-related proteins, S-100B, JNK pathway proteins, and ATG5 were all upregulated, while autophagy-related proteins and s100b were downregulated when the jnk pathway was inhibited or ATG5 was knocked down. ANDRO conferred a survival advantage to hypoxia-injured astrocytes by reducing cell apoptosis and promoting autophagy and s100b expression. Furthermore, the promotion of autophagy and s100b expression by ANDRO was via activation of jnk pathway and regulation of ATG5.

MeSH terms

  • Animals
  • Apoptosis / drug effects
  • Astrocytes / drug effects*
  • Astrocytes / physiology
  • Autophagy / drug effects*
  • Blotting, Western
  • Cell Hypoxia / drug effects*
  • Cell Survival / drug effects
  • Diterpenes / pharmacology*
  • Mice
  • Real-Time Polymerase Chain Reaction
  • S100 Calcium Binding Protein beta Subunit / drug effects*
  • S100 Calcium Binding Protein beta Subunit / metabolism
  • Time Factors
  • Transfection

Substances

  • Diterpenes
  • S100 Calcium Binding Protein beta Subunit
  • S100b protein, mouse
  • andrographolide