Original Article
Modulation of apurinic/apyrimidinic endonuclease-1 on oxidative stress induced pulmonary microvascular endothelial cells injury in rats
Jie Weng, Jinzhen Hou, Luming Tang, Hui Xie, Daqing Chen, Laifang Sun, Binyu Ying, Yuqiang Gong
Published 2016-10-01
Cite as Chin J Crit Care Med(Electronic Edition), 2016, 09(5): 309-314. DOI: 10.3877/cma.j.issn.1674-6880.2016.05.005
Abstract
ObjectiveTo investigate the modulation of apurinic/apyrimidinic endonuclease-1 (APE1) on oxidative stress reaction of pulmonary microvascular endothelial cells (PMVECs) in rats induced by H2O2.
MethodsPMVECs were cultured and randomly divided into 4 groups: control group (no H2O2), H2O2 groups (the final concentrations of H2O2 in cultured cells were 50, 100, 200 μmol/L, respectively). The levels of proliferative activity, reactive oxygen species (ROS) and APE1 were detected at 24 h after the different concentrations of H2O2 stimulation. Meanwhile, APE1 protein expression was checked at 24, 48, 72 h after 100 μmol/mL H2O2 stimulation. Later, the PMVECs were transfected with lentiviruses and randomly divided into 4 groups: control group (no H2O2), H2O2 group (100 μmol/L H2O2), H2O2 + empty vector group (lentivirus vector transfected empty cells with 100 μmol/L H2O2) and H2O2+ APE1 group (APE1 gene transfected lentivirus cells with 100 μmol/L H2O2). The levels of proliferative activity and ROS after 24 h culture were detected and compared.
ResultsCompared with the control group, the levels of proliferative activity, ROS and APE1 in the other three groups increased obviously after being induced by different doses of H2O2 for 24 h (F = 80.238, 445.608, 547.566; all P < 0.05). In the 100 μmol/L H2O2 group, APE1 expression decreased obviously after 0, 24, 48 and 72 h stimulation [(0.781 ± 0.043), (0.611 ± 0.026), (0.407 ± 0.014), (0.272 ± 0.013); all P < 0.05]. After transfection, the level of ROS in the H2O2 group was much higher than that in the control group [(86.4 ± 1.2) vs. (56.4 ± 0.9), P < 0.05]. The level of ROS decreased [(66.8 ± 1.0) vs. (86.4 ± 1.2), P < 0.05] and the level of proliferative activity increased [(0.209 ± 0.001) vs. (0.153 ± 0.001), P < 0.05] obviously in the H2O2 + APE1 group, comparing with the H2O2 group.
ConclusionsH2O2 stimulation can downregulate the APE1 protein expression; conversely, upregulating the APE1 expression increases the level of cell proliferative activity, also inhibit the level of ROS. Thus, APE1 can relieve the oxidative stress reaction of PMVECs in rats induced by H2O2.
Key words:
Hydrogen peroxide; Oxidative stress; Reactive oxygen species; Apurinic/apyrimidinic endonuclease-1; Pulmonary microvascular endothelial cells
Contributor Information
Jie Weng
Departmnet of Emergency, Second Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China
Jinzhen Hou
Departmnet of Emergency, Second Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China
Luming Tang
Departmnet of Emergency, Second Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China
Hui Xie
Departmnet of Emergency, Second Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China
Daqing Chen
Departmnet of Emergency, Second Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China
Laifang Sun
Departmnet of Emergency, Second Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China
Binyu Ying
Department of Intensive Medicine, Second Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China
Yuqiang Gong
Department of Intensive Medicine, Second Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China
