Effects of N-acetylcysteine on oxidative stress, proliferation and apoptosis of HT22 cells induced by radiation
HUANG Yue, CHEN Naiyao, ZHAO Hui, YAN Zhenyu, ZHANG Haixia, ZHAO Xuecong, ZHANG Dingping
RADIATION PROTECTION. 2021, 41(2):
165-173.
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In order to investigate the effect of N-acetyl-L-cysteine (NAC) on radiation-related oxidative stress and the proliferation and apoptosis of hippocampal neuron HT22 cells, the following process was conducted. First, HT22 cells were irradiated with different doses of X-rays (0, 2, 4, 6, 8, 10, 12 Gy), and the optimum radiation dose (10 Gy) was selected. Then experimental groups were set as follows: control (Control) group, radiation treatment (RT) group, RT+NAC group. Continue to cultivate for 24 h after irradiation, CCK-8 was used to detect cell proliferation; The apoptosis of the HT-22 cell was examined by AnnexinV/PI double labeling flow cytometry.Intracellular reactive oxygen species (ROS) was detected by DCFH-DA fluorescent probe to evaluate intracellular oxidative stress levels; Intracellular glutathione (GSH), malondialdehyde (MDA) content and superoxide dismutase (SOD) activity were detected by colorimetry; The expression of Cleaved caspase-3, Bax and BCl-2 protein was analyzed by Western blot. Results show that: (1) The effect of 2 Gy radiation dose on cell proliferation is not obvious. When the radiation dose was higher than 2 Gy, with the increase of radiation dose the proliferation of HT22 cell was decreased (p<0.05). After radiation dose of 10 Gy, cell proliferation inhibition rate was close to 50%. Therefore, 10 Gy was used as the optimal radiation dose for the experiment. (2) Pretreatment with NAC before 10 Gy X-ray irradiation can significantly increase the proliferation rate of HT22 cells. (p<0.01). (3) Radiation treatment can significantly increase intracellular ROS and MDA content (p<0.01), meanwhile intracellular GSH content and SOD activity were reduced (p<0.01). Radiation treatment promoted the expression of apoptotic proteins: Bax and Cleaved caspase-3 (p<0.01). The apoptosis rate was increased significantly (p<0.01). NAC can reduce intracellular ROS and MDA content after irradiation (p<0.01). GSH level and SOD activity were increased (p<0.01). The expression of apoptotic proteins and apoptosis rate were reduced significantly. The above results indicate that NAC can inhibit radiation-related oxidative stress, reduce radiation inhibition of HT22 cell proliferation, and reduce apoptosis.
