Mechanism by which PLD2 alleviates panapoptosis during pancreatic cell injury via Nrf2-NFκB pathway
10.3760/cma.j.cn115807-20240111-00016
- VernacularTitle:PLD2通过Nrf2-NFκB途径缓解胰腺细胞损伤过程中泛凋亡的机制研究
- Author:
Rong ZHANG
1
;
Chaohai WANG
;
Chao DU
Author Information
1. 山西省儿童医院儿童重症医学科，太原 030000
- Keywords:
PLD2;
Nrf2;
Pancreatic cell injury;
Panapoptosis
- From:
Chinese Journal of Endocrine Surgery
2024;18(3):372-376
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the mechanism of PLD2 alleviating panapoptosis during pancreatic cell injury through Nrf2-NFκB pathway.Methods:Rat pancreatic AR42J cells purchased from ATCC were used for experimental study. The cultured and treated cells were divided into the following groups: CON group (AR42J cells cultured under conventional conditions), CER group (AR42J cells were added with 10 nM cerulein in order to establish the in vitro pancreatitis model), CER+pcDNA group (non-target plasmid negative control PcDNA was established on the basis of the in vitro pancreatitis model), and CER+ PLD2-OE group (based on in vitro pancreatitis model, PLD2-OE of PcDNA PLD2-overexpression plasmid was established). PLD2 expression was detected by RT-qPCR. The levels of inflammatory factors in cell supernatant were detected by RT-qPCR. The expression of Nrf2-NFκB signaling pathway was analyzed by protein imprinting. The expressions of apoptosis-related, pyrodeath related and necrotic proteins were analyzed by protein imprinting.Results:The expression of PLD2 mRNA in CER+ PLD2-OE group (1.79±0.12) was higher than that in CON group (0.54±0.01) and CER+pcDNA group (0.62±0.01). The expressions of TNF-α mRNA (2.95±0.21), IL-6 mRNA (2.35±0.18) and IL-10 mRNA (3.22±0.20) in CER+PLD2-OE group were higher than those in CER+ pcDNA group (4.25±0.25; IL-6 mRNA: 3.64±0.21; IL-10 mRNA: 3.22±0.20). The expression of Nrf2 protein (0.49±0.01) in CER group was lower than that in CON group (1.02±0.01), and the expression of NFκB protein (2.52±0.21) in CER group was higher than that in CON group (1.01±0.01). The expression of Nrf2 protein (1.24±0.03) in CER+PLD2-OE group was higher than that in CER+ pcDNA group (0.50±0.01), and the expression of NFκB protein (1.68±0.14) in CER+PLD2-OE group was lower than that in CER+ pcDNA group (2.46±0.22). The expression of Bax protein in CER group (1.83±0.14) was higher than that in CON group (1.04±0.02), and the expression of Bcl-2 protein in CER group (0.31±0.01) was lower than that in CON group (1.02±0.01). The expression of Bcl-2 protein (0.75±0.02) in CER+PLD2-OE group was higher than that in CER+ pcDNA group (0.30±0.01), and the expression of Bax protein (1.42±0.11) in CER+PLD2-OE group was lower than that in CER+ pcDNA group (1.85±0.13). The expression of Gasdermins protein (1.72±0.13), Caspase-1 protein (1.88±0.15) and NLRP3 protein (1.77±0.13) in CER group was higher than that in CON group (Gasdermins: 1.13±0.04; Caspase-1:1.08 ±0.02; NLRP3:1.05±0.03). The expression of Gasdermins protein (1.24±0.05), Caspase-1 protein (1.16±0.04) and NLRP3 protein (1.17±0.05) in CER+PLD2-OE group was higher than that in CER+pcDNA group (Gasdermins: 1.69±0.12; Caspase-1:1.75±0.13; NLRP3:1.80±0.14). The expressions of RIPK1/RIPK3 protein (0.52±0.01), MLKL protein (0.48±0.01) and TNFR1 protein (0.51±0.01) in CER group were higher than those in CON group (RIPK1/RIPK3:1.04 ±0.02; MLKL: 1.03±0.01; TNFR1:1.01±0.01), and CER+PLD2-OE RIPK1/RIPK3 proteins (0.65±0.02), MLKL proteins (0.54±0.01) and TNFR1 proteins (0.63±0.01) were more expressed than CER+pcDNA group (RIPK1/RIPK3: 1.72±0.15; MLKL: 1.65±0.13; TNFR1:1.81±0.16) .Conclusion:PLD2 plays a key role in the regulation of panapoptosis (apoptosis, pyrodeath and necrosis), and effectively alleviates pancreatic cell damage by activating Nrf2 antioxidant pathway and inhibiting NFκB inflammatory pathway.
