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β-conglycinin is a recognized factor in leading to intestinal inflammation and limiting application of soybean meal in aquaculture. Our previous study reported that heat-killed B. siamensis LF4 could effectively mitigate inflammatory response and apoptosis caused by β-conglycinin in spotted seabass (Lateolabrax maculatus) enterocytes, but the mechanisms involved are not fully understood. In the present study, therefore, whole cell wall (CW), peptidoglycan (PG) and lipoteichoic acid (LTA) and cell-free supernatant (CFS) have been collected from B. siamensis LF4 and their mitigative function on β-conglycinin-induced adverse impacts and mechanisms underlying were evaluated. The results showed that β-conglycinin-induced cell injury, characterized with significantly decreased cell viability and increased activities of lactate dehydrogenase, glutamic oxaloacetic transaminase, glutamic propylic transaminase (P < 0.05), were reversed by subsequent heat-killed B. siamensis LF4 and its CW, LTA, PG and CFS treatment. Enterocytes co-cultured with heat-killed B. siamensis LF4 and its CW, LTA, PG and CFS (especially PG) significantly increased expressions of anti-inflammatory genes (IL-2, IL-4, IL-10 and TGF-β1), tight junction proteins (ZO-1, occludin and claudin-b) and antimicrobial peptides (β-defensin, hepcidin-1, NK-lysin and piscidin-5), and decreased expressions of pro-inflammatory genes (IL-1β, IL-8 and TNF-α) and apoptosis-related genes (caspase 3, caspase 8 and caspase 9) (P < 0.05), indicating their excellent mitigation effects on β-conglycinin-induced cell damages. In addition, heat-killed B. siamensis LF4 and its CW, LTA, PG and CFS significantly increased TLR2 mRNA level (especially in PG treatment), and decreased MAPKs (JNK, ERK, p38 and AP-1) and NF-κB related genes expressions. In conclusion, heat-killed B. siamensis LF4 and its CW, LTA, PG and CFS could modulating TLR2/MAPKs/NF-κB signaling and alleviating β-conglycinin-induced enterocytes injury in spotted seabass (L. maculatus), and PG presented the best potential. • B. siamensis LF4 and its CW, LTA, PG and CFS ameliorate β-conglycinin induced inflammation and apoptosis. • B. siamensis LF4 and its CW, LTA, PG and CFS modulate TLR2/MAPKs/NF-κB signaling. • PG showed the best potential in ameliorating β-conglycinin induced inflammation and apoptosis. [ABSTRACT FROM AUTHOR]

β-conglycinin and glycinin, two major heat-stable anti-nutritional factors in soybean meal (SM), have been suggested as the key inducers of intestinal inflammation in aquatic animals. In the present study, a spotted seabass intestinal epithelial cells (IECs) were used to compare the inflammation-inducing effects of β-conglycinin and glycinin. The results showed that IECs co-cultured with 1.0 mg/mL β-conglycinin for 12 h or 1.5 mg/mL glycinin for 24 h significantly decreased the cell viability (P < 0.05), and overstimulated inflammation and apoptosis response by significantly down-regulating anti-inflammatory genes (IL-2, IL-4, IL-10 and TGF-β1) expressions and significantly up-regulated pro-inflammatory genes (IL-1β, IL-8 and TNF-α) and apoptosis genes (caspase 3, caspase 8 and caspase 9) expressions (P < 0.05). Subsequently, a β-conglycinin based inflammation IECs model was established and used for demonstrating whether commensal probiotic B. siamensis LF4 can ameliorate the adverse effects of β-conglycinin. The results showed β-conglycinin-induced cell viability damage was completely repaired by treated with 109 cells/mL heat-killed B. siamensis LF4 for ≥12 h. At the same time, IECs co-cultured with 109 cells/mL heat-killed B. siamensis LF4 for 24 h significantly ameliorated β-conglycinin-induced inflammation and apoptosis by up-regulating anti-inflammatory genes (IL-2, IL-4, IL-10 and TGF-β1) expressions and down-regulated pro-inflammatory genes (IL-1β, IL-8 and TNF-α) and apoptosis genes (caspase 3, caspase 8 and caspase 9) expressions (P < 0.05). In summary, both β-conglycinin and glycinin can lead to inflammation and apoptosis in spotted seabass IECs, and β-conglycinin is more effective; commensal B. siamensis LF4 can efficiently ameliorate β-conglycinin induced inflammation and apoptosis in IECs. • β-conglycinin and glycinin can lead to inflammation and apoptosis in spotted seabass IECs. • β-conglycinin is more effective than glycinin in inducing inflammation and apoptosis. • B. siamensis LF4 can ameliorate β-conglycinin induced inflammation and apoptosis. [ABSTRACT FROM AUTHOR]