6-姜烯酚通过调节CDX2/miR-216a-5p轴缓解棕榈酸诱导的Caco-2细胞紧密连接损伤OA

Palmitic acid(PA)can damage the tight junction of human intestinal epithelial cells,leading to intestinal barrier dysfunction.Our previous study demonstrates that 6-shogaol can modulate the expression of microRNA miR-216a-5p,leading to the targeted inhibition of the Toll-like receptor 4(TLR4)signaling pathway.This,in turn,reduces the production of inflammatory factors and protects intestinal cells from PA-induced damage.However,the specific mechanism by which 6-shogaol regulates miR-216a-5p expression remains unclear.This study establishes a model of intestinal tight junction injury by treating human colorectal adenocarcinoma Caco-2 cells with PA.Through DNA pulldown assays and dual-luciferase reporter gene experiments,this study preliminarily identifies Caudal-related homeobox transcription factor 2(CDX2)as a candidate regulator of miR-216a-5p.By combining chromatin immunoprecipitation followed by polymerase chain reaction(ChIP-PCR)and electrophoretic mobility shift assay(EMSA),this study confirms that the transcription factor CDX2 can bind to the miR-216a-5p promoter and promotes its transcription.Finally,this study evaluates the role of transcription factor CDX2 in regulating the expression of pro-inflammatory factors and tight junction-related proteins,proving that CDX2 is the target through which 6-shogaol regulates miR-216a-5p expression.This study is the first to show that CDX2 is a direct transcriptional activator of miR-216a-5p,providing a reference and basis for revealing the health-regulation effects of ginger and related functional food research.