Protective Effect Of Hemp Seed Polysaccharide On Intestinal Mucosal Injury Induced By Cyclophosphamide In Mice

Hemp seed is a seed of the genus Cannabis sativa L.and has a long history of medicinal use.In this study,we used the water-soluble polysaccharide?HSP?extracted from Hemp seed in this laboratory as the research object,combined with Cy-induced intestinal mucosal injury model in vivo and hydrogen peroxide-induced intestinal epithelial cell IPEC-1 damage model in vitro,to explore the protective effect of Hemp seed polysaccharide on intestinal mucosal injury and its possible mechanism.This provides a theoretical basis for the development of health food related to hemp seed polysaccharides to improve intestinal mucosal damage caused by drugs.Objective:?1?To investigate the protective effect of HSP on oxidative damage of intestinal mucosa induced by Cy in mice?2?To explore the protective effect of HSP on intestinal immunosuppression induced by Cy?3?To investigate the protective effect of HSP on Cy-induced intestinal mucosal injury in mice and its related mechanisms?4?To explore the protective effect of HSP on H₂O₂-induced oxidative damage of IPEC-1 cellsMETHODS:?1?An animal model of intestinal mucosal injury in mice was established by intraperitoneal injection of Cy for four consecutive days.The following indicators were measured:body weight and organ index,thymus,spleen tissue morphology,morphology and ultramicroscopic structure of jejunum,distribution and quantity of goblet cells,intestinal permeability,sIgA content in intestinal lavage fluid,antioxidant factor and cytokine levels in serum,liver and ileal tissue homogenate,and ileal tissue related antioxidant factor and cytokine mRNA expression,ileal tight junction protein ZO-1,Occludin and Claudin-1 and mucin-2 mRNA expression levels and protein expression levels on the Nrf2-Keap1 pathway and NF-?B pathway.?2?An in vitro oxidative stress injury model of IPEC-1 cells was established by using H₂O₂ as an oxidative stressor.The effect of HSP on the proliferation of IPEC-1 cells was detected by MTT assay.The activity of antioxidant enzymes in IPEC-1 cells was measured by enzyme labeling method,and the mRNA expression of related antioxidant enzymes was detected by RT-qPCR.The protein expression level on the Nrf2-Keap1 pathway was detected by Western blot.Conclusion:?1?Pre-administration of HSP has protective effect on Cy-induced intestinal mucosal oxidative damage in mice,and its mechanism may be related to the activation of Nrf2-Keap1 signaling pathway.?2?Pre-administration of HSP has certain immunoregulatory effect on Cy-induced immunosuppression in mice,and its mechanism may be related to the activation of NF-?B pathway.?3?Pre-administration of HSP can protect the intestinal mucosal injury induced by Cy in mice,which may be related to the up-regulation of tight junction protein and mucin.?4?Early administration of HSP0.2 has protective effects on H₂O₂-induced oxidative damage of IPEC-1 cells,and its mechanism may be related to the activation of Keap1/Nrf2 signaling pathway.