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  

Mechanism by which PLD2 alleviates panapoptosis during pancreatic cell injury via Nrf2-NFκB pathway

作　　者：张荣 王朝海[1] 杜超 Zhang Rong;Wang Chaohai;Du Chao(Department of Pediatric Critical Care Medicine,Shanxi Provincial Children’s Hospital,Taiyuan 030000,China)

机构地区：[1]山西省儿童医院儿童重症医学科,太原030000

出　　处：《中华内分泌外科杂志（中英文）》2024年第3期372-376,共5页Chinese Journal of Endocrine Surgery

基　　金：山西省儿童医院院级课题(2021050)。

摘　　要：目的探讨磷脂酰胆碱特异性磷脂酶D2(phosphatidylcholine specific phospholipase D2,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。方法将大鼠胰腺AR42J细胞用于实验研究,采用随机数字法分为CON组(常规条件下培养的AR42J细胞)、CER组(建立体外胰腺炎模型,在AR42J细胞中加入10 nM cerulein)、CER+pcDNA组(在体外胰腺炎模型的基础上,建立非靶质粒阴性对照PcDNA)、CER+PLD2-OE组(在体外胰腺炎模型的基础上,建立了PcDNA的PLD2过表达质粒PLD2-OE)。通过RT-qPCR检测PLD2的表达。通过RT-qPCR检测细胞上清液炎症因子的水平。通过蛋白印迹分析Nrf2-NFκB信号通路的表达。通过蛋白印迹分析凋亡相关、焦亡相关和坏死相关蛋白的表达。结果CER组(0.54±0.01)和CER+pcDNA组(0.62±0.01)PLD2 mRNA表达较CON组(1.02±0.03)降低,CER+PLD2-OE组(1.79±0.12)PLD2 mRNA表达较CON组升高。CER+PLD2-OE组的TNF-αmRNA(2.95±0.21)、IL-6 mRNA(2.35±0.18)和IL-10 mRNA(3.22±0.20)表达较CER+pcDNA组(TNF-αmRNA:4.25±0.25、IL-6 mRNA:3.64±0.21、IL-10 mRNA:3.22±0.20)降低。CER组Nrf2蛋白(0.49±0.01)表达较CON组(1.02±0.01)降低,CER组NFκB蛋白(2.52±0.21)表达较CON组(1.01±0.01)升高。CER+PLD2-OE组Nrf2蛋白(1.24±0.03)表达较CER+pcDNA组(0.50±0.01)升高,CER+PLD2-OE组NFκB蛋白(1.68±0.14)表达较CER+pcDNA组(2.46±0.22)降低。CER组Bax蛋白(1.83±0.14)表达较CON组(1.04±0.02)升高,CER组Bcl-2蛋白(0.31±0.01)表达较CON组(1.02±0.01)降低。CER+PLD2-OE组Bcl-2蛋白(0.75±0.02)表达较CER+pcDNA组(0.30±0.01)升高,CER+PLD2-OE组Bax蛋白(1.42±0.11)表达较CER+pcDNA组(1.85±0.13)降低。CER组Gasdermins蛋白(1.72±0.13)、Caspase-1蛋白(1.88±0.15)和NLRP3蛋白(1.77±0.13)表达较CON组(Gasdermins:1.13±0.04、Caspase-1:1.08±0.02、NLRP3:1.05±0.03)升高,CER+PLD2-OE组Gasdermins蛋白(1.24±0.05)、Caspase-1蛋白(1.16±0.04)和NLRP3蛋白(1.17±0.05)表达较CER+pcDNA组(Gasdermins:1.69±0.12、Caspase-1:1.75±0.13Objective 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-αmRNA(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(0.7

关 键 词：磷脂酰胆碱特异性磷脂酶D2 核因子红细胞相关因子2 胰腺细胞损伤 泛凋亡 

分 类 号：R576[医药卫生—消化系统]