Department of General Surgery, Xuzhou Children's Hospital, Xuzhou Medical University, China.
Department of General Surgery, Children's Hospital of Soochow University, China.
Biomed Pharmacother. 2020 May;125:109867. doi: 10.1016/j.biopha.2020.109867. Epub 2020 Feb 12.
To investigate the relationship between FOXO3 overexpression and NLRP3 and explore the effect of FOXO3 on necrotizing colitis.
100 clean grade newborn SD (Sprague Dawley) rats were randomly divided into 4 groups: NEC group, NEC + FOXO3a group, NEC + NC group and control group. NEC rat model was established by hypoxia + hypothermia stimulation; HE staining was used for detection of the inflammation of intestinal tissue. The histological scores of intestinal tissues were histologically scored, generally, there were three types of inflammatory scoring systems including anatomically based systems, severity-based systems and quality of life systems (Lim et al., 2015) and in this study we utilized severity-based systems by HE staining. Human intestinal epithelial cell line was transfected with recombinant plasmid overexpressing FOXO3a and recombinant plasmid overexpressing NLRP3, and divided into control group, LPS group, LPS + NC group, LPS + FOXO3a group and LPS + FOXO3a + NLRP3 group; Caspase-1 was used for the detection of pyroptosis. The expressions of FOXO3a, NLRP3, cleaved Caspase-1 and the expression of TLR4 in TLR4 signaling pathway were detected by RT-qPCR and WB. IL-1β, IL-6, IL-18 and TNF-α were detected by ELISA.
(1) FOXO3a is under-expressed and NLRP3 is highly expressed in NEC neonatal rat intestinal tissue. (2) The inflammatory condition of intestinal tissue in NEC + FOXO3a group was improved compared with NEC group (P < 0.05). (3) FOXO3a was highly expressed in NEC + FOXO3a group. The expression of IL-1β, IL-6, IL-18, SOD and MDA in NEC + FOXO3a group was lower than that in NEC group. (4) The expression of IL-1β, IL-6, IL-18, SOD and MDA in intestinal epithelial cells of LPS + FOXO3a group was lower than other groups. (5) Overexpression of FOXO3a inhibits LPS-induced pyroptotic cell death in intestinal epithelial cells by inhibiting NLRP3.
Overexpression of FOXO3 in mice with necrotizing colitis can improve inflammatory conditions in mice by affecting NLRP3-mediated cell caking.
探讨 FOXO3 过表达与 NLRP3 的关系，并研究 FOXO3 对坏死性结肠炎的影响。
100 只清洁级新生 SD（Sprague Dawley）大鼠随机分为 4 组：NEC 组、NEC+FOXO3a 组、NEC+NC 组和对照组。采用缺氧+低温刺激建立 NEC 大鼠模型；采用 HE 染色检测肠组织炎症情况。对肠组织的组织学评分进行组织学评分，一般来说，有三种基于解剖的炎症评分系统、基于严重程度的系统和基于生活质量的系统（Lim 等人，2015），本研究采用基于 HE 染色的严重程度系统。将人肠上皮细胞系转染过表达 FOXO3a 的重组质粒和过表达 NLRP3 的重组质粒，分为对照组、LPS 组、LPS+NC 组、LPS+FOXO3a 组和 LPS+FOXO3a+NLRP3 组；用 Caspase-1 检测细胞焦亡。采用 RT-qPCR 和 WB 检测 TLR4 信号通路中 FOXO3a、NLRP3、cleaved Caspase-1 的表达及 TLR4 的表达。采用 ELISA 检测 IL-1β、IL-6、IL-18 和 TNF-α。
（1）NEC 新生大鼠肠组织中 FOXO3a 表达下调，NLRP3 表达上调。（2）与 NEC 组相比，NEC+FOXO3a 组肠组织炎症情况改善（P<0.05）。（3）NEC+FOXO3a 组 FOXO3a 高表达，NEC+FOXO3a 组 IL-1β、IL-6、IL-18、SOD 和 MDA 表达低于 NEC 组。（4）LPS+FOXO3a 组肠上皮细胞中 IL-1β、IL-6、IL-18、SOD 和 MDA 的表达均低于其他各组。（5）过表达 FOXO3a 通过抑制 NLRP3 抑制 LPS 诱导的肠上皮细胞焦亡。
坏死性结肠炎小鼠中 FOXO3 的过表达可通过影响 NLRP3 介导的细胞焦亡来改善小鼠的炎症状态。
