Aloin suppresses lipopolysaccharide-induced inflammation by inhibiting JAK1-STAT1/3 activation and ROS production in RAW264.7 cells

Abstract

The anti-inflammatory effects of aloin, a bioactive ingredient extracted from Aloe vera, have been described previously. The present study aimed to assess these effects and explore the underlying molecular mechanisms. RAW264.7 cells were incubated with different doses of aloin (100, 150 and 200 µg/ml) and lipopolysaccharide (LPS; 100 ng/ml) for the indicated times. Then, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 expression levels were detected by western blot analysis and reverse transcription polymerase chain reaction (RT-PCR). The concentrations of inflammatory cytokines in the cell culture supernatant were determined by ELISA. Total nitric oxide (NO) assay and reactive oxygen species (ROS) kits were used to detect NO and ROS levels, respectively. Mitogen-activated protein kinase, nuclear factor κB and Janus kinase-signal transducer and activator of transcription (JAK-STAT) pathway activation were verified by western blot analysis. Confocal and nucleocytoplasmic separation experiments were used to detect STAT nuclear translocation. It was identified that aloin decreased the level of LPS-induced iNOS expression, inhibiting the release of interleukin (IL)-1β, IL-6, tumour necrosis factor-α and NO dose-dependently. Mechanistically, aloin suppressed LPS-induced JAK1-STAT1/3 activation and STAT1/3 nuclear translocation. Additionally, LPS-induced ROS production was inhibited by aloin. Collectively, these data suggest that aloin attenuated LPS-induced inflammation by inhibiting ROS-mediated activation of the JAK1-STAT1/3 signalling pathway, thereby inhibiting the nuclear translocation of STAT1/3 in RAW264.7 cells. The present study provides an experimental basis for the clinical application of aloin in inflammatory-associated diseases.