摘要
目的:研究巯基丙酮酸硫基转移酶(MPST)介导PI3K/AKT信号对急性胰腺炎(AP)细胞模型凋亡和自噬的影响.方法:构建AP体外细胞模型,分组为 CON组、AP组、AP + siMPST组和 AP + oeMPST组.采用 CCK-8 法检测细胞活力;采用ELISA法检测细胞上清炎症因子TNF-α、IL-1 和IL-6 水平;采用流式细胞仪检测细胞凋亡水平;采用Western blotting检测腺泡细胞LC3 Ⅱ/Ⅰ、beclin1、ATG5、MPST、PI3K、p-PI3K、AKT、p-AKT的表达水平.结果:AP组、AP +siMPST组和AP +oeMPST组的细胞存活率均低于 CON 组(P<0.01),而细胞凋亡率则均高于 CON 组(P<0.01);AP + siMPST 组细胞存活率、AP + oeMPST组细胞凋亡率均高于AP组(P<0.01),而AP +siMPST组细胞凋亡率、AP +oeMPST组细胞存活率均低于AP组(P<0.01).AP组、AP +oeMPST组的腺泡细胞TNF-α、IL-1 和IL-6、LC3 Ⅱ/Ⅰ、beclin1、ATG5、MPST、p-PI3K/PI3K、p-AKT/AKT水平明显高于CON组(P<0.01);上述指标水平AP +siMPST组均低于AP组(P<0.01),而AP +oeMPST组则均高于AP组(P<0.01).结论:MPST可通过PI3K/AKT信号诱导胰腺腺泡细胞凋亡、自噬和炎症反应;抑制MPST可能对AP具有治疗意义.
Abstract
Objective:To investigate the effect of mercaptopyruvate thiotransferase(MPST)mediated PI3K/AKT signaling on apoptosis and autophagy in acute pancreatitis(AP)cell model.Methods:An AP in vitro cell model was constructed.Cells were divided into CON group,AP group,AP +siMPST group,and AP +oeMPST group.CCK-8 method was used to detect cell viability.The levels of inflammatory cytokines TNF-α,IL-1 and IL-6 were detected by ELISA.Flow cytometry was used to detect the level of apoptosis.Western blotting was used to detect the expression levels of LC3 Ⅱ/Ⅰ,beclin1,ATG5,MPST,PI3K,p-PI3K,AKT,and p-AKT in acinar cells.Results:The cell survival rates of the AP group,AP + siMPST group,and AP + oeMPST group were all lower than those of the CON group(P<0.01),while the apoptosis rate was higher than that of the CON group(P<0.01);The cell survival rate of the AP + siMPST group and the apoptosis rate of the AP +oeMPST group were higher than those of the AP group(P<0.01),while the apoptosis rate of the AP +siMPST group and the cell survival rate of the AP +oeMPST group were lower than those of the AP group(P<0.01).The levels of TNF-α,IL-1,IL-6,LC3 Ⅱ/Ⅰ,beclin1,ATG5,MPST,p-PI3K/PI3K,and p-AKT/AKT in acinar cells of AP group and AP +oeMPST group were significantly higher than in the CON group(P<0.01).The above indicators were lower in the AP +siMPST group than in the AP group(P<0.01),while the AP + oeMPST group was higher than in the AP group(P<0.01).Conclusions:MPST can induce pancreatic acinar cell apoptosis,autophagy,and inflammatory response through PI3K/AKT signaling.Inhibiting MPST may have therapeutic significance for AP.
基金项目
江苏省扬州市重点研发项目(社会发展)(YZ2021091)
江苏省扬州市"绿扬金凤计划"卫生创新领军人才基金项目(YZLYJF2020WSCX037)
江苏省中医药科技发展计划项目(YB2020088)
NETL
NSTL
维普
万方数据