N-乙酰半胱氨酸对HT22细胞辐射诱导氧化应激及增殖和凋亡的影响

Abstract

Abstract: 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.

Key words: ionizing radiation, HT22, oxidative stress, NAC, apoptosis

CLC Number:

R142+.4

HUANG Yue, CHEN Naiyao, ZHAO Hui, YAN Zhenyu, ZHANG Haixia, ZHAO Xuecong, ZHANG Dingping. Effects of N-acetylcysteine on oxidative stress, proliferation and apoptosis of HT22 cells induced by radiation[J].RADIATION PROTECTION, 2021, 41(2): 165-173.

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Effects of N-acetylcysteine on oxidative stress, proliferation and apoptosis of HT22 cells induced by radiation

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