ABSTRACT
Objective To clone the murine Na+-K+-2Cl-cotransporter (Nkcc2) gene promoter and analyze 20-HETE regulation of the murine Nkcc2 gene transcriptional activity. Methods A fragment of the murine Nkcc2 gene promoter was analyzed using bioinformatics software AliBaba and TRANSFAC TESS. The murine Nkcc2 gene promoter fragment (-1 462 bp-+40 bp) was amplified by PCR using murine genomic DNA as a template and then cloned into a pGL3-Basic vector to generate a luciferase reporter construct. The recombinant reporter construct was transiently transfected into HEK293 T cells using Lipofectamine 2000 for 24 h. The transfected HEK293 T cells were treated with 20-HETE for 2 h followed by measurement of the luciferase activity using the Dual-Luciferase Reporter Assay system. Results A luciferase reporter construct containing the murine Nkcc2 gene promoter was successfully generated. The results showed that 20-HETE significantly reduced the transcriptional activity of the construct. Conclusion 20-HETE may reduce the expression of the murine Nkcc2 gene through transcriptional regulation.
ABSTRACT
Chronic atrophic gastritis (CAG) is a common and refractory disease of the digestive system in clinic.Apoptosis is the important reason which relates CAG pathological changes.As one of the important means of clinical prevention and treatment of digestive diseases, traditional Chinese medicine (TCM) in the role and mechanism of CAG related research has become an important direction in recent years.Based on the collection of related literatures and data, the paper analyzed the possible mechanism of CAG pathogenesis grasped the key aspect of apoptosis and clarified the research status of TCM on the regulation of gastric mucosal cell apoptosis in CAG in recent years through focusing on the specific signaling pathways in mitochondrial pathway, death receptor pathway, endoplasmic reticulum pathway.Through the above analysis, the paper provided some ideas for further systematic and in-depth research and explored CAG effective TCM program.