Aloperine alleviates LPS-induced inflammation in bovine intestinal epithelial cells through autophagy and TLR4/p38 MAPK/NF-κB pathway

Calf diarrhea is a major cause of mortality and morbidity, leading to substantial economic losses in the cattle industry. Aloperine (Alo) exhibits an anti-inflammation effect and alleviates dextran sulfate sodium salt (DSS)-induced colitis; however, it remains unclear whether Alo alleviates calf diarrhea-induced intestinal inflammation. In this study, network pharmacology was used to discover the possible mechanism of Alo on the anti-inflammatory effect; Then, an inflammation model was induced by LPS in BIECs-21 to evaluate the protective effect of Alo on LPS-induced inflammation. Results found that a total of 68 overlapping targets of Alo and inflammation were obtained, among which ALB, AKT1, IL-6, and EGFR exhibited good affinity for Alo. In vitro experiments, Alo inhibited LPS-induced pro-inflammatory cytokine levels, increased the expression of ZO-1 and Claudin 1, and reduced the expression of proteins related to autophagy and TLR4/p38 MAPK/NF-κB pathway; the autophagy inhibitor CQ and Baf A1 results further demonstrated that Alo inhibited late stages of autophagy maturation and impaired intestinal epithelial barrier function. These results indicated that Alo has protective effects on LPS-induced inflammation by decreasing the levels of the pro-inflammatory cytokines through the TLR4/p38 MAPK/NF-κB pathway, inhibiting late stages of autophagy maturation, and impairing intestinal epithelial barrier function.