The regulation and mechanism of thioredoxin 1 and thioredoxin interacting protein on nucleotide-binding oligomerized domain-like receptor protein 3 inflammasome in peritoneal mesothelial cells
10.3760/cma.j.cn115455-20230223-00174
- VernacularTitle:硫氧还蛋白1/硫氧还蛋白相互作用蛋白对腹膜间皮细胞核苷酸结合寡聚化结构域样受体蛋白3炎症小体的调控及机制研究
- Author:
Rui CHENG
1
;
Changcai ZHU
Author Information
1. 黄石市爱康医院泌尿外科，黄石 435000
- Keywords:
Peritoneal mesothelial cells;
Thioredoxin 1;
Thioredoxin interacting protein;
Nucleotide binding oligomerization domain-like receptor protein 3
- From:
Chinese Journal of Postgraduates of Medicine
2024;47(6):513-517
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the regulatory effect and possible mechanism of thioredoxin 1 (Trx1)/thioredoxin interacting protein (Txnip) on nucleotide-binding oligomerized domain-like receptor protein 3 (NLRP3) inflammasome in peritoneal mesothelial cells.Methods:Human peritoneal mesothelial cell line (HMrSV5) was divided into three groups by random number table method. One group was cultured with 10% fetal bovine serum (FBS) medium (the blank group), another group was cultured with 4.25% peritoneal dialysate (PDS) and 10% FBS medium with transfected small interfering RNA (siRNA) negative control (si-NC group), the last group was transfected with 4.25% PDS and 10% FBS culture medium (si-Txnip group). Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting (WB) were used to determine the mRNA and protein expressions of Trx1, Txnip and NLRP3 in each group, respectively; MTT method was used to determine cell proliferation in each group at different time points. The levels of interleukin (IL)-1β, IL-6 and the activity of caspase-1 in the supernatant were determined by enzyme-linked immunosorbent assay (ELISA).Results:The proliferative activity of HMrSV5 cultured for 1, 2 and 3 d in si-NC group and si-Txnip group were lower than those in the blank group ( P<0.05); the proliferative activity of HMrSV5 cultured for 1, 2 and 3 d in si-Txnip group were higher than those in the si-NC group : 0.402 ± 0.009 vs. 0.372 ± 0.010, 0.554 ± 0.016 vs. 0.482 ± 0.008, 0.700 ± 0.013 vs. 0.590 ± 0.010, there were statistical differences ( P<0.05). The mRNA expressions of Txnip and NLRP3 in si-NC group and si-Txnip group were higher than those in the blank group ( P<0.05), and the mRNA expressions of Trx1 and NLRP3 in si-Txnip group were lower than those in the si-NC group: 1.43 ± 0.32 vs. 2.80 ± 0.43, 2.38 ± 0.35 vs. 3.42 ± 0.40; the mRNA expression of Trx1 in si-Txnip group were lower than that in the blank group and higher than that in the si-NC group: 2.24 ± 0.35 vs. 3.50 ± 0.38 and 1.18 ± 0.23, there were statistical differences ( P<0.05). The Txnip and NLRP3 protein expressions in the si-NC group and si-Txnip group were higher than those in blank group ( P<0.05). The expression of Txnip and NLRP3 protein in the si-Txnip group was lower than that in the si-NC group: 0.453 ± 0.108 vs. 0.754 ± 0.116; the expression of Trx1 protein in the si-Txnip group was lower than that in blank group and higher than that in si-NC group: 0.514 ± 0.112 vs. 0.753 ± 0.125 and 0.297 ± 0.010, there were statistical differences ( P<0.05). The levels of interleukin(IL)-1β and IL-6 in supernatant in the si-NC group and si-Txnip group were higher than those in the blank group: (0.45 ± 0.07), (0.35 ± 0.06) μg/L vs. (0.23 ± 0.05) μg/L, (3.00 ± 0.38), (2.32 ± 0.30) μg/L vs. (1.95 ± 0.34) μg/L, there were statistical differences ( P<0.05). The levels of IL-1β and IL-6 of supernatant in the si-Txnip group were lower than those in the si-NC group, there were statistical differences ( P<0.05). The caspase-1 activity of supernatant in the si-Txnip group was lower than that in the si-NC group and higher than that in the blank group: (0.87 ± 0.26) U vs. (1.65 ± 0.40), (0.62 ± 0.20) U, there were statistical differences ( P<0.05). Conclusions:Down-regulation of Txnip in Trx1/Txnip system can significantly inhibit the NLRP3 inflammasome and its function in peritoneal mesothelial cells.
