Andrograpanin mitigates lipopolysaccharides induced endometritis via TLR4/NF-κB pathway

Endometritis is an inflammatory disease that is caused by various pathogenic organisms. Andrograpanin is a compound of Andrographis paniculata, which has an important role in many inflammatory diseases, but the molecular mechanism of andrograpanin to combat inflammation is unclear. This study shows the anti-inflammatory effect of andrograpanin on Lipopolysaccharides (LPS) stimulated bovine endometrial epithelial cells (bEECs) and LPS-induced mouse model. We investigated the cytotoxic effect of bEECs by using CCK-8 analysis. Quantification of pro-inflammatory cytokines (IL-1 beta, IL-6, and TNF-alpha) protein levels and mRNA was carried out using RT-qPCR and ELISA, respectively. The protein expressions of p65 and I kappa B alpha were assessed by western blot and immunofluorescence to check the inhibition of p65 translocation into the nucleus. The treatment effect of andrograpanin on mouse uterine tissues was determined by histopathology. in vivo, curative effect experiments showed that andrograpanin significantly reduced the endometrial injury in a mouse model. Our studies first confirmed that andrograpanin had no cytotoxic effect at 7.5,15 and 30 mu g/mL concentration on bEECs. Following, Andrograpanin significantly reduced the mRNA and protein level of IL-1 beta, IL-6, and TNF-alpha both in vivo and in vitro. Finally, Andrograpanin inhibited the I kappa B alpha degradation and p65 phosphorylation in LPS-stimulated bEECs and LPS-induced endometrial injury. Our results showed that andrograpanin might have therapeutic effects against endometritis. (C) 2022 Society for Biology of Reproduction & the Institute of Animal Reproduction and Food Research of Polish Academy of Sciences in Olsztyn. Published by Elsevier B.V. All rights reserved.