摘要
目的探讨P22phox对缺氧诱导的人肺动脉平滑肌细胞(HPASMCs)的作用及其可能的作用机制。方法体外常氧或者缺氧培养HPASMCs,将HPASMCs随机分为空白对照组、si-NC+oe-NC组、si-P22phox+oe-NC组和si-P22phox+oe-KLF4组,按照分组进行细胞转染,随后进行缺氧培养。CCK-8实验检测细胞活力;EdU实验检测细胞增殖能力;细胞划痕实验检测细胞迁移;Transwell实验检测细胞迁移;qRT-PCR和Western blot检测细胞P22phox和KLF4的表达。结果与常氧组相比,缺氧组细胞活力、EdU^(+)细胞数、划痕愈合率和迁移细胞数显著升高,P22phox和KLF4 mRNA表达以及P22phox和KLF4蛋白表达显著升高(均P<0.05)。与空白对照组相比,si-NC+oe-NC组细胞各指标差异无统计学意义(P>0.05);与si-NC+oe-NC组相比,si-P22phox+oe-NC组细胞P22phox和KLF4 mRNA表达显著降低,P22phox和KLF4蛋白表达显著降低,细胞活力、EdU^(+)细胞数、划痕愈合率和迁移细胞数显著降低(均P<0.05);与si-P22phox+oe-NC组相比,si-P22phox+oe-KLF4组细胞P22phox mRNA和蛋白表达差异无统计学意义(P>0.05),KLF4 mRNA和蛋白表达显著升高,细胞活力、EdU^(+)细胞数、划痕愈合率和迁移细胞数显著升高(均P<0.05)。结论P22phox通过上调KLF4的表达,促进缺氧诱导的HPASMCs增殖和迁移。
Objective To explore the effect of P22phox on human pulmonary artery smooth muscle cells(HPASMCs)induced by hypoxia and its possible mechanism.Methods HPASMCs were cultured under normoxia or hypoxia in vitro.HPASMCs were randomly divided into blank control group,si-NC+oe-NC group,si-P22phox+oe-NC group and si-P22phox+oe-KLF4 group,The cells are transfected according to the grouping and then cultured under hypoxia.CCK-8 experiment was used to detect cell viability.EdU experiment was used to detect cell proliferation ability.Cell scratch experiment was used to detect cell migration.Transwell experiment was used to detect cell migration.qRT-PCR and Western blot were used to detect the expression of P22phox and KLF4 in cells.Results Compared with the normoxic group,the cell viability,EdU+cell number,scratch healing rate and number of migrating cells in the hypoxia group were significantly increased(P<0.05),and the expression of P22phox and KLF4 mRNA and the expression of P22phox and KLF4 protein were significantly increased(P<0.05).Compared with the blank control group,the si-NC+oe-NC group had no significant difference in cell indicators(P>0v05).Compared with the si-NC+oe-NC group,the expression of P22phox and KLF4 mRNA in si-P22phox+oe-NC group was significantly reduced(P<0.05),the expression of P22phox and KLF4 protein was significantly reduced(P<0.05),cell viability,EdU+cell number,scratch healing rate and number of migrating cells were significantly reduced(P<0.05).Compared with the si-P22phox+oe-NC group,the expression of P22phox mRNA and protein in the si-P22phox+oe-KLF4 group was not statistically different(P>0.05),while the expression of KLF4 mRNA and protein was significantly increased(P<0.05),cell viability,EdU+cell number,scratch healing rate and number of migrating cells were significantly increased(P<0.05).Conclusion P22phox promotes the proliferation and migration of HPASMCs induced by hypoxia by up-regulating the expression of KLF4.
作者
古丽那扎尔
李倩
米娜瓦尔·胡加艾合买提
吴雷琪
阿加尔古丽·依米提
Gulinazhaer;LI Qian;MINAWAER Hujiaaihemaiti;WU Leiqi;AJIAERGULI·Yimiti(Department of General Medicine, The First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, China)
出处
《西部医学》
2022年第1期33-39,共7页
Medical Journal of West China
基金
新疆维吾尔自治区自然科学基金(2019D01C013)。
