Interleukin-4 affects microglial autophagic flux

Interleukin-4 plays an important protective role in Alzheimer's disease by regulating microglial phenotype, phagocytosis of amyloid-beta, and secretion of anti-inflammatory and neurotrophic cytokines. Recently, increasing evidence has suggested that autophagy regulates innate immunity by affecting M1/M2 polarization of microglia/macrophages. However, the role of interleukin-4 in microglial autophagy is unknown. In view of this, BV2 microglia were treated with 0, 10, 20 or 50 ng/mL interleukin-4 for 24, 48, or 72 hours. Subsequently, light chain 3-II and p62 protein expression levels were detected by western blot assay. BV2 microglia were incubated with interleukin-4 (20 ng/mL, experimental group), 3-methyladenine (500 mu M, autophagy inhibitor, negative control group), rapamycin (100 nM, autophagy inductor, positive control group), 3-methyladenine + interleukin-4 (rescue group), or without treatment for 24 hours, and then exposed to amyloid-beta (1 mu M, model group) or vehicle control (control) for 24 hours. LC3-II and p62 protein expression levels were again detected by western blot assay. In addition, expression levels of multiple markers of M1 and M2 phenotype were assessed by real-time fluorescence quantitative polymerase chain reaction, while intracellular and supernatant amyloid-beta protein levels were measured by enzyme-linked immunosorbent assay. Our results showed that interleukin-4 induced microglial autophagic flux, most significantly at 20 ng/mL for 48 hours. Interleukin-4 pretreated microglia inhibited blockade of amyloid-beta-induced autophagic flux, and promoted amyloid-beta uptake and degradation partly through autophagic flux, but inhibited switching of amyloid-beta-induced M1 phenotype independent on autophagic flux. These results indicate that interleukin-4 pretreated microglia increases uptake and degradation of amyloid-beta in a process partly mediated by autophagy, which may play a protective role against Alzheimer's disease.