title: The anti-inflammatory effects of asiatic acid in lipopolysaccharide-stimulated human corneal epithelial cells.
abstract: OBJECTIVE
  To investigate the anti-inflammatory effects of asiatic acid (AA) on lipopolysaccharide (LPS)-induced inflammatory response in human corneal epithelial cells (HCECs).
  
  
  METHODS
  Cell viability was measured using a cell counting kit-8 (CCK-8) assay. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to determine the mRNA expression of interleukin-8 (IL-8), interleukin-6 (IL-6), interleukin-1β (IL-1β), tumor necrosis factor-alpha (TNF-α), and transforming growth factor-β (TGF-β) in HCECs. Intracellular reactive oxygen species (ROS) was measured using the ROS assay kit. Glutathione (GSH) concentration was measured using the total GSH assay kit. Akt1 and Akt phosphorylation (p-Akt1) levels were measured by Western blotting and immunofluorescence.
  
  
  RESULTS
  AA induced toxicity at high concentrations and significantly stimulated the proliferation of HCECs at concentrations of 20 µmol/L for 1h. LPS at concentrations of 300 ng/mL for 1h significantly stimulated the mRNA expression of IL-8, IL-6, IL-1β, TNF-α, and TGF-β in HCECs, while the stimulation effects were significantly inhibited by AA (20 µmol/L). In addition, AA was found to decrease the content of ROS, increase GSH generation, and also inhibit LPS-induced p-Akt in HCECs.
  
  
  CONCLUSIONS
  AA decreases the generation of inflammatory factors IL-8, IL-6, IL-1β, TNF-α, and TGF-β in LPS-stimulated HCECs. AA significantly inhibites the intracellular concentrations of ROS and increases GSH generation. AA also inhibites LPS-induced p-Akt in HCECs. These findings reveal that AA has anti-inflammation effects in LPS-stimulated HCECs.
authors.affiliation: Special Medicine Department of Medical College, Qingdao University, Qingdao 266071, Shandong Province, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Chen
authors.affiliation: Special Medicine Department of Medical College, Qingdao University, Qingdao 266071, Shandong Province, China.
authors.forename: Xiao-Min
authors.initials: XM
authors.lastname: Hua
authors.affiliation: Special Medicine Department of Medical College, Qingdao University, Qingdao 266071, Shandong Province, China.
authors.forename: Bai-Chen
authors.initials: BC
authors.lastname: Ze
authors.affiliation: Special Medicine Department of Medical College, Qingdao University, Qingdao 266071, Shandong Province, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Wang
authors.affiliation: Special Medicine Department of Medical College, Qingdao University, Qingdao 266071, Shandong Province, China.
authors.forename: Li
authors.initials: L
authors.lastname: Wei
keywords: Akt phosphorylation; asiatic acid; glutathione; inflammatory factors; lipopolysaccharide; reactive oxygen species