Alpha-linolenic acid based nano-suspension protect against lipopolysaccharides induced mastitis by inhibiting NFκBp65, HIF-1α, and mitochondria-mediated apoptotic pathway in albino Wistar rats

Toxicol Appl Pharmacol. 2019 Aug 15:377:114628. doi: 10.1016/j.taap.2019.114628. Epub 2019 Jun 14.


The purpose of this study was to investigate the therapeutic effects and underlying mechanism of alpha-linolenic acid based intra-mammary nano-suspension (ALA-NS) on both in vitro antimicrobial and in vivo activity. The ALA-NS formulated and optimized for parameters like particle size, zeta potential, polydispersity index, sedimentation volume, and stability studies. In vitro, our results showed that ALA-NS (F1 and F2) have the higher zone of inhibition and lower minimum inhibitory concentration (MIC) value than ALA and cefotaxime alone against mastitis-causing pathogens. In vivo, our results showed that ALA-NS (F1 and F2) restored the altered oxidative biomarkers (superoxide dismutase, catalase, glutathione, TBARs, and protein carbonyl) along with histopathological changes in lipopolysaccharides (LPS) treated rats. Western blot results indicated that ALA-NS (F1 and F2) inhibited LPS induced inflammatory proteins (NFκBp65, COX, LOX, and IFN-γ) in rat mammary epithelial cells. ALA-NS (F1 and F2) also suppressed the hypoxia inducible factor-1α (HIF-1α) and upregulated prolyl-hydroxylase (PHD-2), sterol regulatory element binding protein (SREBP-1c), and fatty acid synthase (FASN) protein expression. In addition, ALA-NS upregulated the pro-apoptotic (BAX and BAD) and downregulated anti-apoptotic (BCL-2 and BCL-XL) proteins expression in rat mammary epithelial tissue. In conclusion, we found that ALA-NS (F1 and F2) have in vitro antimicrobial activity and protective effects on LPS-induced mastitis in rats.

Keywords: Alpha-linolenic acid; Hypoxia; Inflammation; Lipopolysaccharides; Mastitis.

MeSH terms

  • Animals
  • Anti-Infective Agents / pharmacology
  • Apoptosis / drug effects*
  • Apoptosis Regulatory Proteins / drug effects
  • Bacteria / drug effects
  • Epithelial Cells / drug effects
  • Female
  • Hypoxia-Inducible Factor 1, alpha Subunit / antagonists & inhibitors*
  • Lipopolysaccharides / antagonists & inhibitors*
  • Lipopolysaccharides / toxicity
  • Mammary Glands, Animal / cytology
  • Mammary Glands, Animal / drug effects
  • Mastitis / chemically induced*
  • Mastitis / microbiology
  • Mastitis / prevention & control*
  • Microbial Sensitivity Tests
  • Mitochondria / drug effects*
  • Oxidative Stress / drug effects
  • Rats
  • Rats, Wistar
  • Signal Transduction / drug effects
  • Suspensions
  • Transcription Factor RelA / antagonists & inhibitors*
  • alpha-Linolenic Acid / administration & dosage
  • alpha-Linolenic Acid / pharmacology*


  • Anti-Infective Agents
  • Apoptosis Regulatory Proteins
  • Hif1a protein, rat
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • Lipopolysaccharides
  • Suspensions
  • Transcription Factor RelA
  • alpha-Linolenic Acid