NOX4在幽门螺旋杆菌诱导胃癌细胞ROS中的作用

doi:10.6040/j.issn.1671-7554.0.2021.1348

摘要/Abstract

摘要： 目的研究NOX4在幽门螺旋杆菌(H.pylori)诱导胃癌细胞氧化应激中的作用。方法将胃癌细胞株MKN-45分别与幽门螺旋杆菌菌株NCTC 26695、NCTC 11637共培养6 h,实验设未感染组与幽门螺旋杆菌感染组;用25 mmol/L的活性氧清除剂N-乙酰半胱氨酸(NAC)预处理MKN-45细胞30 min,实验设NAC未处理组与NAC处理组。用Western blotting检测细胞内还原型烟酰胺腺嘌呤二核苷酸磷酸氧化酶 4(NOX4)蛋白表达水平;采用活性氧检测试剂盒检测细胞内活性氧(ROS)水平,荧光显微镜观察幽门螺旋杆菌感染后ROS水平的变化,荧光酶标仪检测荧光强度。结果与未感染组相比较,幽门螺旋杆菌感染组细胞内ROS生成增多;感染NCTC 26695后NOX4蛋白的表达增加(t=11.67,P<0.001),感染NCTC 11637后,NOX4蛋白的表达增加(t=17.29,P<0.001);无论是NCTC 26695还是NCTC 11637菌株,NOX4蛋白表达上调的差异均有统计学意义(P<0.05)。NAC预处理后再用幽门螺旋杆菌感染,细胞中NOX4的表达显著降低［NCTC 26695(P<0.001);NCTC 11637(P<0.001)］。结论 NOX4/ROS途径可能参与幽门螺旋杆菌对胃癌细胞氧化还原信号的激活。

关键词: 活性氧, 还原型烟酰胺腺嘌呤二核苷酸磷酸氧化酶 4, N-乙醇半胱氨酸, 胃癌, 氧化还原信号

Abstract: Objective To investigate the effect of nicotinamide adenine dinucleotide phosphate oxidase 4(NOX4)on reactive oxygen species(ROS)induced by Helicobacter pylori(H. pylori)in gastric cancer cells. Methods Gastric cancer cell line MKN-45 was co-cultured with H. pylori NCTC 26695 and NCTC 11637 for 6 hours respectively, the experiment included non-infection group and H. pylori infection group; MKN-45 cells were pretreated with 25mmol/L N-acetyl cysteine(NAC)for 30 minutes, NAC treated group and NAC untreated group were set up. The protein expression of NOX4 was detected with Western blotting. The level of ROS was detected with ROS detection kit. Changes in ROS level after H. pylori infection were observed with fluorescence microscope, and fluorescence intensity was detected with fluorescein microplate analyzer. Results ROS production increased in H. pylori infected group compared with non-infection group. NOX4 protein expression increased after NCTC 26695 infection(t=11.67, P<0.001), and increased after NCTC 11637 infection(t=17.29, P<0.001). After NAC pretreatment and H. pylori infection, NOX4 expression was significantly reduced ［NCTC 26695(P<0.001), NCTC 11637(P<0.001)］. Conclusion NOX4/ROS pathway may be involved in the activation of H. pylori redox signal in gastric cancer cells.

Key words: Reactive oxygen species, Nicotinamide adenine dinucleotide phosphate oxidase 4, N-acetylcysteine, Gastric cancer, Redox signal

中图分类号:

R735.2

包舒晴,杨明月,刘端瑞,汪运山,郏雁飞. NOX4在幽门螺旋杆菌诱导胃癌细胞ROS中的作用[J]. 山东大学学报 (医学版), 2022, 60(6): 19-25.

BAO Shuqing, YANG Mingyue, LIU Duanrui, WANG Yunshan, JIA Yanfei. Role of NOX4 in ROS induction by Helicobacter pylori in gastric cancer cells[J]. Journal of Shandong University (Health Sciences), 2022, 60(6): 19-25.

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