摘要
目的:探究核受体相互作用蛋白1(NRIP1)通过转录调控高迁移率族蛋白B1(HMGB1)对脓毒症引发的肠道上皮细胞损伤的作用。方法:RT-qPCR和Western blot检测NRIP1和HMGB1表达水平。HE染色检测大鼠肠道组织的病理学变化;CCK-8法确定LPS的最佳处理时间。利用NRIP1小干扰RNA(siRNA-NRIP1-1/2)转染Caco-2细胞,采用CCK-8法和流式细胞术分别检测细胞活力和凋亡;RT-qPCR和Western blot分析炎症相关因子表达;跨上皮电阻(TEER)检测肠道上皮通透性;Western blot检测凋亡和紧密连接相关的蛋白表达。荧光素酶报告实验和染色质免疫沉淀法(ChIP)验证NRIP1与HMGB1的结合关系。在LPS处理的Caco-2细胞中同时敲低NRIP1并过表达HMGB1后再次进行功能实验。结果:NRIP1在脓毒症大鼠肠道组织和LPS诱导的Caco-2细胞中表达增加。干扰NRIP1后抑制LPS诱导的Caco-2细胞活性损伤、凋亡、炎症反应和屏障损伤。此外,NRIP1可转录激活HMGB1表达,过表达HMGB1可逆转NRIP1缺失对Caco-2细胞活力、凋亡、炎症反应以及屏障功能的影响。结论:NRIP1可促进脓毒症引发的肠道上皮细胞损伤,其机制可能与HMGB1的转录激活相关。
Objective:To investigate the impacts of nuclear receptor-interacting protein 1(NRIP1)on sepsis-evoked intestinal epithelial injury via transcriptional regulation of high mobility group box 1(HMGB1).Methods:The expression levels of NRIP1 and HMGB1 were detected by RT-qPCR and Western blot.The pathological changes of intestinal tissue were detected by HE staining.CCK-8 assay determined the optimal treatment time of LPS.Caco-2 cells were transfected with NRIP1 small interfering RNA(siRNA-NRIP1-1/2),and cell viability and apoptosis were detected by CCK-8 assay and flow cytometry,respectively.RT-qPCR and Western blot examined the expressions of inflammation-associated factors.Transepithelial resistance(TEER)was used to detect intestinal epithelial permeability.Western blot was used to detect the expressions of apoptosis and tight-junction related proteins.The binding relationship between NRIP1 and HMGB1 was verified by luciferase reporting assay and chromatin immunoprecipitation assay(ChIP).After knocking down NRIP1 and overexpressing HMGB1 in LPS-treated Caco-2 cells,the functional experiment was performed again.Results:NRIP1 expression was fortified in the intestinal tissues of sepsis rats and LPS-treated Caco-2 cells.Interference with NRIP1 attenuated LPS-elicited Caco-2 cell viability injury,apoptosis,inflammatory response and barrier damage.Additionally,NRIP1 might activate HMGB1 expression at transcriptional level and HMGB1 elevation might reverse the impacts of NRIP1 absence on Caco-2 cell viability,apoptosis,inflammatory response as well as barrier function.Conclusion:NRIP1 may promote sepsis-elicited intestinal epithelial injury,which may be related to transcriptional activation of HMGB1.
作者
崔文娟
刘芹
樊晓光
乔鲁军
CUI Wenjuan;LIU Qin;FAN Xiaoguang;QIAO Lujun(Intensive Care Unit,Shengli Oilfield Central Hospital,Dongying 257000,China)
出处
《中国免疫学杂志》
北大核心
2025年第2期328-335,共8页
Chinese Journal of Immunology
基金
山东省医药卫生科技发展计划项目,黄河三角洲学者专项经费资助(202117010420)。
作者简介
崔文娟,女,硕士,副主任医师,主要从事重症医学研究,E-mail:yixuefeimeng@163.com;通信作者:乔鲁军,男,硕士,主任医师,主要从事重症医学研究,E-mail:qiaolujun@sina.cn。
