[1]安雨轩,肖宗懿,王小燕,等.A549细胞氧糖剥夺/恢复模型中焦亡与凋亡的共享信号通路[J].陆军军医大学学报(原第三军医大学学报),2022,44(11):1119-1125.
 AN Yuxuan,XIAO Zongyi,WANG Xiaoyan,et al.Shared signaling pathway in pyroptosis and apoptosis in A549 cells induced by oxygen and glucose deprivation/recovery[J].J Amry Med Univ (J Third Mil Med Univ),2022,44(11):1119-1125.
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
44卷
期数:
2022年第11期
页码:
1119-1125
栏目:
基础医学
出版日期:
2022-06-15

文章信息/Info

Title:
Shared signaling pathway in pyroptosis and apoptosis in A549 cells induced by oxygen and glucose deprivation/recovery
作者:
安雨轩肖宗懿王小燕宋娟陈菲王寿勇
重庆医科大学附属儿童医院麻醉科,国家儿童健康与疾病临床医学研究中心,儿童发育疾病研究教育部重点实验室,儿科学重庆市重点实验室
 
Author(s):
AN Yuxuan XIAO Zongyi WANG Xiaoyan SONG Juan CHEN Fei WANG Shouyong

Department of Anesthesiology, National Clinical Research Center for Child Health and Disorders, Key Laboratory of Child Development and Disorders of Ministry of Education, Chongqing Key Laboratory of Pediatrics, Children’s Hospital of Chongqing Medical University, Chongqing, 400014, China

关键词:
氧糖剥夺细胞焦亡细胞凋亡A549肺泡上皮细胞
Keywords:
oxygen-glucose deprivation pyroptosis apoptosis hypotriploid alveolar basal epithelial A549 cells 
分类号:
R322.35; R329.25; R329.26
文献标志码:
A
摘要:

目的研究氧糖剥夺/恢复(oxygen-glucose deprivation/recovery, OGD/R)对A549肺泡上皮细胞凋亡与焦亡的作用,并探讨二者之间的相互影响。方法培养人肺泡A549细胞株,建立氧糖剥夺/恢复细胞损伤模型,分为对照组(control),氧糖剥夺/恢复组(OGD/R),氧糖剥夺/恢复复合焦亡抑制剂组(OGD/R+VX-765),氧糖剥夺/恢复复合凋亡抑制剂组(OGD/R+FMK),采用流式细胞术检测细胞凋亡率,采用ELISA法测定培养细胞上清中焦亡标志物IL-1β、IL-18水平,采用RT-qPCR和Western blot法测定细胞凋亡和焦亡关键分子的mRNA与蛋白质表达水平。结果与对照组比较,OGD/R组A549细胞凋亡率升高[(3.67±0.64)与(12.57±1.97),P<0.05]。与OGD/R组比较,OGD/R+VX-765组凋亡率降低[(5.97±1.28),P<0.05];与对照组比较,OGD/R组培养上清中焦亡标志物IL-1β、IL-18水平升高[(7.80±0.80)ng/L、(7.99±0.11) ng/L与(17.35±0.44) ng/L、(18.06±1.73) ng/L,P<0.05];与OGD/R组比较,OGD/R+FMK组IL-1β、IL-18水平降低[(4.72±0.59)ng/L、(10.24±1.42)ng/L,P<0.05];OGD/R可刺激A549细胞凋亡与焦亡关键分子的mRNA和蛋白表达;与OGD/R组比较,OGD/R+VX-765组BID的mRNA和蛋白相对表达量降低[(1.74±0.14)、(1.29±0.09) vs (1.12±0.18)、(1.01±0.10),P<0.05],OGD/R+FMK组GSDME的mRNA和蛋白相对表达量降低[(1.35±0.10)、(1.20±0.06) vs (0.94±0.05)、(0.52±0.03),P<0.05]。结论OGD/R可同时引发A549细胞发生焦亡与凋亡,且细胞焦亡与凋亡存在共享信号通路;两种抑制剂均可同时抑制OGD/R引起的A549细胞焦亡与凋亡。

Abstract:

ObjectiveTo investigate the effects of oxygen and glucose deprivation/recovery (OGD/R) on apoptosis and pyroptosis of A549 cells, and to explore the possibility of the mutual regulation in apoptosis and pyroptosis. MethodsA549 cells were resuscitated and cultured, randomly divided into control group, OGD/R group, OGD/R+pyroptosis inhibitor group (OGD/R+VX-765) and OGD/R+apoptosis inhibitor group (OGD/R+FMK). The apoptotic rate was detected by flow cytometry, the levels of pyroptosis markers IL-1β and IL-18 in the supernatant were measured by ELISA, and the expression of key molecules in pyroptosis and apoptosis at mRNA and protein levels were detected by RT-qPCR and Western blotting, respectively. ResultsThe apoptotic rate was significantly increased in the OGD/R group than the control group (12.57±1.97 vs 3.67±0.64, P<0.05), and the rate of the OGD/R+VX-765 group was decreased than that of the OGD/R group (5.97±1.28, P<0.05). OGD/R resulted in obvious increased levels of IL-1β and IL-18 in the supernatant (17.35±0.44 vs 7.80±0.80 ng/L, 18.06±1.73 vs 7.99±0.11 ng/L, P<0.05), but FMK treatment notably reversed the increases of 2 inflammatory factors (4.72±0.59, 10.24±1.42 ng/L, P<0.05). OGD/R enhanced the mRNA and protein expression levels of key molecules of apoptosis and proptosis in A549 cells. While, VX-765 decreased the mRNA and protein levels of BID (1.12±0.18 vs 1.74±0.14, 1.01±0.10 vs 1.29±0.09, P<0.05), and FMK reduced those of GSDME (0.94±0.05 vs 1.35±0.10, 0.52±0.03 vs 1.20±0.06, P<0.05) when compared with the cells from the OGD/R group. ConclusionOGD/R can induce pyroptosis and apoptosis in A549 cells simultaneously, and there exists shared signaling pathway in pyroptosis and apoptosis. Pyroptosis and apoptosis inhibitors can simultaneously inhibit the 2 types of OGD/R-induced cell death.

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更新日期/Last Update: 2022-06-08