Effects of Protocatechuic Acid on Necroptosis and Toll Like Receptor 4 Signaling Pathway in Porcine Small Intestinal Epithelial Cells Induced by Enterotoxigenic Escherichia coli K88
This experiment was conducted to study the effects of protocatechuic acid(PCA)on necroptosis and Toll like receptor 4(TLR4)signaling pathway in porcine small intestinal epithelial cells(IPEC-1 cells)induced by enterotoxigenic Escherichia coli K88(ETEC K88).The experiment used a 2×2 factor design,di-vided into four groups:control group,PCA group(40 µmol/L PCA treated 24 h),ETEC K88 group(50 times the number of cells ETEC K88 treated 2 h)and PCA+ETEC K88 group(40 µmol/L PCA treated 24 h+50 times the number of cells ETEC K88 treated 2 h),each group contained 3 replicates.The results showed as follows:1)compared with the control group,the cell viability of ETEC K88 group was significantly decreased(P＜0.05),and the cell supernatant lactate dehydrogenase(LDH)activity was significantly increased(P＜0.05);compared with the ETEC K88 group,the cell viability of PCA+ETEC K88 group was significantly in-creased(P＜0.05),and the cell supernatant LDH activity was significantly decreased(P＜0.05).2)Com-pared with the control group,ETEC K88 treatment resulted in morphological damage and ultrastructural de-struction of cells,such as cell membrane rupture,nuclear shrinkage,chromatin overflow,swelling and vacuo-lation of mitochondria,leading to an increase in cell necrosis rate;the cell necrosis rate of ETEC K88 group was significantly increased(P＜0.05).Compared with the ETEC K88 group,adding PCA can protect cell morphology;the cell necrosis rate of PCA+ETEC K88 group was significantly decreased(P＜0.05).3)Com-pared with the control group,the mRNA relative expression levels of tumor necrosis factor-α(TNF-α),TLR4,lipopolysaccharide binding protein(LBP),myeloid differentiation protein 2(MD2),interleukin recep-tor-related kinase 1(IRAK1),tumor necrosis factor receptor-related factor 6(TRAF6)and myeloid differenti-ation factor 88(MyD88)of ETEC K88 group were significantly increased(P＜0.05);compared with the ETEC K88 group,the mRNA relative expression levels of TNF-α,TLR4,LBP,MD2,IRAK1,TRAF6 and MyD88 of PCA+ETEC K88 group were significantly decreased(P＜0.05).4)Compared with the control group,the mRNA relative expression levels of tumor necrosis factor receptor 1(TNFR1),Fas associated death domain(FADD),mixed lineage kinase like domain protein(MLKL),high mobility protein 1(HMGB1),dynamin related protein 1(Drp1)and phosphoglycerate mutase 5(PGAM5)of ETEC K88 group were signif-icantly increased(P＜0.05);compared with the ETEC K88 group,the mRNA relative expression levels of TNFR1,FADD,HMGB1,Drp1 and PGAM5 of PCA+ETEC K88 group were significantly decreased(P＜0.05).In conclusion,PCA may alleviate the inflammatory reaction and injury of IPEC-1 cells induced by ETEC K88 by inhibiting activation of necroptosis and TLR4 signaling pathway.[Chinese Journal of Animal Nutrition,2024,36(7):4665-4676]
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维普
