Abstract: SA-PO754
Effect of Hypoxia-Inducible Factor-1α Stabilizers on the Destruction of Tight Junctions in Uremic Intestinal Epithelial Cells
Session Information
- CKD: Mechanisms - III
November 09, 2019 | Location: Exhibit Hall, Walter E. Washington Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: CKD (Non-Dialysis)
- 2103 CKD (Non-Dialysis): Mechanisms
Author
- Liu, Hua, the First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi, China
Background
Hypoxia-inducible factor-1α (HIF-1α) plays an important role in maintaining the structure and function of the intestinal mucosal barrier and is involved in regulating the expression of intestinal epithelial tight junction proteins. In this study, based on the previous findings, the role of HIF-1α in intestinal barrier dysfunction caused by ureima toxins and the effect of intervention of HIF-1α stabilizer (FG-4592) on the above effects were explored.
Methods
Caco2 cells was cultured and divided into 3 groups, the normal control group, the uremic serum stimulation group, and the uremic serum + HIF-1α stabilizer (FG-4592) intervention group. RT-qPCR and Western blotting method was used to detect the expression of HIF-1α, ZO-1, claudin-1, Occludin, and NLRP3 inflammasome in each group. RT-qPCR was used to detect the effect of miR-223 mimimc and siNLRP3 on NLRP3 expression stimulated by uremia serum. Transfection of miR-223-mimics or miR-223-inhibitor to Caco2 cells followed by downstream gene expression analysis is performed to elucidate the targets and roles of mir-223.
Results
Compared with the control group, the application of uremia serum to Caco2 cells can significantly decrease the expression of HIF-1α, ZO-1, claudin-1, occludin mRNA and protein (P < 0.05), while FG-4592 intervention could reverse the above changes (P < 0.05) .On the contrary, stimulation of Caco2 cells by uremia serum can significantly increase the expression of NLRP3, caspase-1 and IL-1β mRNA and protein, while FG-4592 intervention can down-regulate the expression of NLRP3 inflammasome. At the same time, while uremic serum stimulation can increase NLRP3 expression, application of miR-223 mimic or siNLRP3 can reduce NLRP3 expression (P < 0.05). The mRNA and protein expression of NLRP3, caspase-1 and IL-1β of Caco2 cells transfected miR-223-mimic decreased but that increased for tight junction protein, while transfection of miR-223-inhibitor, we got opposite results.
Conclusion
HIF-1α stabilizer (FG-4592) increases the decrease of HIF-1α expression under the stimulation of uremic serum, and its improvement of the expression mechanism of tight junction protein may play a role in the down-regulation of NLRP3 inflammasome caused by the up-regulation of miR-223 expression, thus improving the intestinal barrier function, which needs further experimental verification