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Abstract: SA-PO746

The Expression of Intestinal Mir-223/NLRP3 Axis in Uremic Rats and the Intervention of Probiotics

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

Authors

  • Liu, Hua, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi, China
  • Chen, Lei, Xi'an Jiaotong University, Xi'an, China
  • Liang, Shanshan, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, China
  • Wei, Meng, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, China
  • Sun, Lingshuang, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, China
  • Xue, Jinhong, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi, China
  • Wang, Meng, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, China
  • Jiang, Hongli, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, China
Background

To investigate the role of chronic inflammatory response mediated by NLRP3 inflammasome in uremia intestinal barrier dysfunction by detecting the changes of mir-223 /NLRP3, and to observe the effect of probiotic intervention on it.

Methods

Seventy SD rats were randomly divided into 3 groups and 20 sham group (SH group). 5/6 Nephrectomy was performed on 50 rats in the uremic model group, and the final histopathology confirmed the successful modeling, which were randomly divided into the uremic group (UR group, n=20), probiotics intervention group (UP group, n=20), and intestinal tissue were extraction. The expression of miR-223 mRNA of each group was detected by RT-qPCR. Expression of NLRP3, caspase-1, IL-1β, tight junction protein in intestinal epithelial cells were detected by Western blotting. Bioinformatics software was used to predict the binding site of mir-223-3p and NLRP3 gene.The 3'UTR sequence of NLRP3 gene and its mutants were cloned into the dual luciferase reporter gene vector, and the wild-type and mutant recombinant double luciferase reporter plasmid vector were constructed. PCR and gene sequencing were used to determine whether the vector was successfully constructed, and 293 cells were used for grouping transfection and cell luciferase detection.

Results

Compared with SH group, the expression of miR-223 mRNA in the ileal tissues of the UR group was significantly decreased, while that of UP group was significantly increased (P<0.05). Western blotting showed that the expressions of NLRP3, caspase-1 and IL-1β of the UR group were significantly higher than those of the SH group (P<0.05), while the expressions of JAM-1, Occludin and claudin-1 proteins were significantly lower (P<0.05). However, after probiotics intervention, the expressions of NLRP3, caspase-1 and IL-1b was decreased (P<0.05), and the expressions of JAM-1, Occludin and claudin-1was improved. After 293 cells co-transfected with miR-223-3p mimic or inhibitor, NLRP3-3 'UTR wild-type had decreased or increased luciferase activity, with statistically significant differences (P < 0.05).

Conclusion

Probiotics can improve the intestinal barrier function of uremia, which may involve the miR-223 /NLRP3 pathway. The conclusion of this study may provide a new treatment idea for the intestinal barrier dysfunction of uremia and chronic inflammation.