收稿日期: 2023-11-25
网络出版日期: 2024-07-30
基金资助
国家自然科学基金项目(82160903);江西省自然科学基金青年基金项目(20212BAB216060);江西省中医药中青年骨干人才（第四批）培养计划(赣中医药科教字［2022］7号)
The effect of miR⁃143⁃3p on pyroptosis of ulcerative colitis cells by regulating TLR2/NF⁃κB/NLRP3
Received date: 2023-11-25
Online published: 2024-07-30
目的 研究miR-143-3p对LPS+ATP诱导HCT-116细胞焦亡的影响。 方法 双荧光素酶检测miR-143-3p与TLR2基因靶向关系。转染miR-143-3p mimic后建立焦亡模型，流式细胞术检测细胞凋亡，生化法检测Caspase-1活性及LDH含量；ELISA法检测IL-1β、IL-18含量。q-PCR检测NF-κB、TLR2、NLRP3、GSDMD mRNA水平；免疫荧光检测NLRP3、ASC蛋白表达；Western blot检测TLR2、GSDMD、p-NF-κB p65、cleave Caspase-1蛋白表达。 结果 双荧光素酶检测发现miR-143-3p靶向调控TLR2表达。miR-143-3p mimic组和si TLR2组较模型组细胞中NF-κB、TLR2、NLRP3、GSDMD mRNA表达及TLR2、GSDMD、p-NF-κB p65、cleave Caspase-1、ASC、NLRP3蛋白表达降低；Caspase-1活性及LDH、IL-1β、IL-18含量降低。 结论 miR-143-3p靶向TLR2基因调控NF-κB/NLRP3/Caspase-1信号通路调控溃疡性结肠炎细胞焦亡。
关键词： 溃疡性结肠炎; 细胞焦亡; miR-143-3p; TLR2; TLR2/ NF-κB/NLRP3
史秀丽 , 陈嘉琪 , 朱璠 , 曾娟 , 吴娜 . miR-143-3p靶向调控TLR2/ NF-κB/NLRP3通路对溃疡性结肠炎细胞焦亡的影响[J]. 实用医学杂志, 2024 , 40(15) : 2056 -2062 . DOI: 10.3969/j.issn.1006-5725.2024.15.003
Objective To study the effect of miR-143-3p on pyroptosis of HCT-116 cells induced by LPS + ATP. Methods The targeting relationship between miR-143-3 p and TLR2 gene was detected by dual luciferase. The pyroptosis model was established after transfection of miR-143-3p mimic. Cell apoptosis was detected by flow cytometry， and the activity of Caspase-1 and LDH was detected by biochemical method. The levels of IL-1β and IL-18 were detected by ELISA. The mRNA levels of NF-κB， TLR2， NLRP3 and GSDMD were detected by q-PCR. The expression of NLRP3 and ASC was detected by immunofluorescence. The protein expressions of TLR2， GSDMD， p-NF-κB p65 and cleave Caspase-1 were detected by Western blot. Results Dual luciferase assay showed that miR-143-3p targeted TLR2 expression. The expressions of NF-κB， TLR2， NLRP3， GSDMD mRNA and TLR2， GSDMD， p-NF-κB p65， cleave Caspase-1， ASC， NLRP3 protein in miR-143-3p mimic group and si TLR2 group were lower than those in model group. The activity of Caspase-1 and the content of LDH， IL-1β and IL-18 decreased. Conclusion miR-143-3p regulates pyroptosis of ulcerative colitis cells by targeting TLR2 gene and regulating NF-κB/NLRP3/Caspase-1 signaling pathway.
Key words： ulcerative colitis; cell pyroptosis; miR-143-3p; TLR2; TLR2/ NF-κB/NLRP3
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