Heat stress-induced intestinal epithelial cells necroptosis via TLR3-TRIF-RIP3 pathway was dependent on p53

作者：Zhou, Junjie; Qin, Xihe; Li, Li; Tian, Dan; Zou, Zhimin; Gu, Zhengtao^*; Su, Lei^*

来源：International Immunopharmacology, 2023, 122: 110574.

DOI：10.1016/j.intimp.2023.110574

• Heat stress
• Heatstroke
• P53
• Necroptosis
• Intestine epithelial cells

Background: Heatstroke is a life-threatening disease. Present study was aimed to investigate the mechanism in heat induced intestinal epithelial cell death.Method: Heat stress in vitro model was established on IEC cells with 42 & DEG;C for 2 h. Caspase-8 inhibitor, Caspase-3 inhibitor, RIP3 inhibitor, TLR3 agonist, poly(I:C) and p53 knockdown were used to determine the signaling pathway. Heatstroke in vivo model was established on C57BL/6 mice, with a temperature of 35.5 & DEG;C & PLUSMN;0.5 & DEG;C and a relative humidity of 60% & PLUSMN; 5%. The intestine necroptosis and inflammatory cytokines were measured. Pifithrin & alpha; (3 mg/kg) and p53 knockout mice were used to evaluate the role of p53.Results: Heat stress-induced reduction of cell viability was remarkable reversed by RIP3 inhibitor. Heat stress induced upregulation of TLR3 and facilitate the formation of TRIF-RIP3 complex. The heat stress induced upregulation of RIP3 and p-RIP3 were normalized by the deletion of p53. Meanwhile, p53 knockout decreased TLR3 expression and blocked the formation of TLR3-TRIF complex. The deletion of p53 blocked the decreased cell viability and restored the activation of RIP3-MLKL signaling after heat stress, however, which were abolished by re-expression of p53 via Tp53 OE. Increased the expression of TLR3 in the p53-deficient cells could not affect the heat stress induced necrotic cell death, which suggests that heat stress induced necroptosis via TLR3-TRIFRIP3 signaling pathway is dependent on p53.Conclusion: Heat stress promoted p53 phosphorylation, then upregulated TLR3 and enhanced the interaction of TRIF-RIP3, which would activate the RIP3-MLKL signaling pathway to mediate necroptosis in intestinal epithelial cells.

• 单位
  南方医科大学; 1