Wang Lin, Zhang Lin
Department of Orthopedics, Yijishan Hospital, Wannan Medical College, Wuhu 241000, Anhui, China.
Hangzhou TCM Hospital Affiliated to Zhejiang Chinese Medical University, Hangzhou 310053, Zhejiang, China.
Evid Based Complement Alternat Med. 2022 Sep 21;2022:2483669. doi: 10.1155/2022/2483669. eCollection 2022.
To investigate the role and mechanism of protein kinase N2 (PKN2) in hydrogen peroxide (HO)-induced injury of PC12 cells.
s. PC12 cells were transfected with lentivirus to knock down or overexpress PKN2 and then were treated with 300 M HO to establish a cell model of oxidative stress injury. The cell viability of PC12 cells in each group was determined by the CCK-8 method. Biochemical assays were used to measure reactive oxygen species (ROS), malondialdehyde (MDA) levels, and superoxide dismutase (SOD) activity. Western blot was used to detect the protein expressions of PKN2, caspase-3, cleaved-caspase-3, PARP, cleaved-PARP, p-mTOR, and mTOR in PC12 cells in each group.
HO treatment could significantly reduce PC12 cell viability and promote cell apoptosis and oxidative stress. PKN2 overexpression inhibited HO-induced apoptosis and oxidation damage by increasing PC12 cell viability, SOD activity, and p-mTOR protein expression, reducing intracellular ROS and MDA levels, and cleaved-caspase-3 and cleaved-PARP protein expression.
PKN2 overexpression can alleviate HO-induced oxidative stress injury and apoptosis in PC12 cells by activating the mTOR pathway.
探讨蛋白激酶N2（PKN2）在过氧化氢（H₂O₂）诱导的PC12细胞损伤中的作用及机制。
用慢病毒转染PC12细胞以敲低或过表达PKN2，然后用300μM H₂O₂处理以建立氧化应激损伤细胞模型。采用CCK-8法测定各组PC12细胞的活力。生化检测用于测量活性氧（ROS）、丙二醛（MDA）水平和超氧化物歧化酶（SOD）活性。蛋白质印迹法用于检测各组PC12细胞中PKN2、半胱天冬酶-3（caspase-3）、裂解的半胱天冬酶-3、聚（ADP-核糖）聚合酶（PARP）、裂解的PARP、磷酸化的哺乳动物雷帕霉素靶蛋白（p-mTOR）和mTOR的蛋白表达。
H₂O₂处理可显著降低PC12细胞活力，促进细胞凋亡和氧化应激。PKN2过表达通过提高PC12细胞活力、SOD活性和p-mTOR蛋白表达，降低细胞内ROS和MDA水平以及裂解的半胱天冬酶-3和裂解的PARP蛋白表达，抑制H₂O₂诱导的细胞凋亡和氧化损伤。
PKN2过表达可通过激活mTOR通路减轻H₂O₂诱导的PC12细胞氧化应激损伤和细胞凋亡。
