Department of Ultrasound, The Affiliated Hospital of Yunnan University (The Forth Affiliated Hospital of Kunming Medical University, The Second People's Hospital of Yunnan Province), Kunming, Yunnan, China.
Department of Vascular Surgery, The Affiliated Hospital of Yunnan University (The Forth Affiliated Hospital of Kunming Medical University, The Second People's Hospital of Yunnan Province), Kunming, Yunnan, China.
J Immunol Res. 2022 Aug 19;2022:6909764. doi: 10.1155/2022/6909764. eCollection 2022.
This study investigated whether vascular endothelial necroptosis is involved in deep vein thrombosis (DVT) and how IL-17B facilitates necroptosis signaling.
The DVT mouse model was induced by ligation of the IVC. The cross-sectional area of thrombus increases and the thrombus occupied the entire venous lumen at 48 h after ligation. Meanwhile, the increased expression of p-RIP3/RIP3 was most pronounced at 48 h after ligation, and the p-MLKL/MLKL peaked at 72 h.
Based on Illumina sequencing and KEGG pathway analyses, the activated RIP3/MLKL is associated with increased IL-17B. With thrombus formation, IL-17B was upregulated and enhanced the expression of RIP3 and MLKL in the IVC wall, as well as their phosphorylation levels (all < 0.05, the comparison group consisted of the control group, DVT group, DVT/IL-17B group, and DVT/anti-IL-17B group). The p-RIP3/RIP3 and p-MLKL/MLKL ratios were reduced by anti-IL-17B. Similarly, the weight and cross-sectional area of the thrombi were increased by IL-17B and decreased by the IL-17B antibody. IL-17B had a smaller effect on thrombosis in knockout mice compared with WT mice. In vitro, the IL-17B protein expression and the level of RIP3 and MLKL phosphorylation increased high in the OGD cells, accompanied by increased expression of IL-6 and TNF-. IL-17B enhanced the expression of IL-6 and TNF- but had little effect on the IL-6 and TNF- after transfected with siRIP3 or siMLKL. Similarly, the plasma IL-17B, IL-6, and TNF- were significantly increased after thrombosis in WT mice, and enhanced by IL-17B. But IL-17B did not increase the plasma IL-6 and TNF- in knockout mice.
In conclusion, those results suggest that vascular endothelial necroptosis plays a crucial role in vascular injury and IL-17B could enhance the necroptosis pathway.
本研究旨在探讨血管内皮细胞坏死是否参与深静脉血栓形成（DVT），以及 IL-17B 如何促进坏死信号通路。
通过结扎 IVC 诱导 DVT 小鼠模型。结扎后 48 小时，血栓的横截面积增加，血栓占据整个静脉腔。同时，结扎后 48 小时 RIP3/RIP3 的磷酸化表达最明显，p-MLKL/MLKL 在 72 小时达到峰值。
基于 Illumina 测序和 KEGG 通路分析，激活的 RIP3/MLKL 与 IL-17B 的增加有关。随着血栓形成，IL-17B 上调，并增强 IVC 壁中 RIP3 和 MLKL 的表达及其磷酸化水平（均 < 0.05，对照组由对照组、DVT 组、DVT/IL-17B 组和 DVT/抗 IL-17B 组组成）。抗 IL-17B 降低了 p-RIP3/RIP3 和 p-MLKL/MLKL 比值。同样，IL-17B 增加血栓的重量和横截面积，而 IL-17B 抗体则减少血栓。与 WT 小鼠相比，IL-17B 在 knockout 小鼠中对血栓形成的影响较小。在体外，IL-17B 蛋白表达和 RIP3 和 MLKL 磷酸化水平在 OGD 细胞中升高，同时 IL-6 和 TNF-α表达增加。IL-17B 增强了 IL-6 和 TNF-α的表达，但转染 siRIP3 或 siMLKL 后对 IL-6 和 TNF-α的影响不大。同样，WT 小鼠血栓形成后血浆 IL-17B、IL-6 和 TNF-α显著升高，IL-17B 增强。但 IL-17B 不会增加 knockout 小鼠的血浆 IL-6 和 TNF-α。
综上所述，这些结果表明血管内皮细胞坏死在血管损伤中起关键作用，IL-17B 可增强坏死通路。
