P7C3-A20对创伤性脑损伤的PC12细胞凋亡和氧化的影响

doi:10.11843/j.issn.0366-6964.2022.12.028

摘要/Abstract

摘要： 旨在研究3,6-二溴-beta-氟-N-(3-甲氧基苯基)-9H-咔唑-9-丙胺(P7C3-A20)对大鼠肾上腺髓质嗜铬瘤分化细胞株(PC12细胞)创伤性脑损伤(TBI)的修复作用。将细胞分为对照组(A组)、模型组(B组)、0.03 μmol·L^-1药物治疗组(C组)、0.3 μmol·L^-1药物治疗组(D组)、3 μmol·L^-1药物治疗组(E组)和药物空白组(F组)，TBI细胞模型通过使用10 μL枪头划出横竖相间4 mm的直线来制备。使用CCK8试剂盒检测细胞活力，荧光显微镜观察细胞凋亡、活性氧(ROS)情况，荧光定量PCR仪检测半胱氨酸蛋白酶-3(Caspase-3)、B细胞淋巴瘤/白血病-2基因(Bcl-2)、血红素氧合酶1(HO-1)、NAD (P) H:醌氧化还原酶1(NQO1)和谷氨酸半胱氨酸连接酶催化亚基(GCLC)的mRNA相对表达量。结果显示：TBI造模后细胞活力极显著降低(P<0.01)，0.03 μmol·L^-1浓度的P7C3-A20能显著提高TBI后的细胞活力(P<0.05)，0.3 μmol·L^-1浓度的P7C3-A20能极显著提高TBI后的细胞活力(P<0.01)。TBI造模后细胞早晚期凋亡细胞比例极显著增加(P<0.01)，0.03 μmol·L^-1浓度的P7C3-A20能显著减少TBI后的早期凋亡细胞比例(P<0.05)，极显著减少TBI后的晚期凋亡细胞比例(P<0.01)，0.3和3 μmol·L^-1浓度的P7C3-A20能极显著减少TBI后的早晚期凋亡细胞比例(P<0.01)。TBI造模后活性氧细胞比例极显著增加(P<0.01)，0.03和3 μmol·L^-1浓度的P7C3-A20能显著减少TBI后的活性氧细胞比例(P<0.05)，0.3 μmol·L^-1浓度的P7C3-A20能极显著减少TBI后的活性氧细胞比例(P<0.01)。0.3 μmol·L^-1浓度的P7C3-A20能极显著减少TBI后Caspase-3的mRNA相对表达量(P<0.01)，显著提升TBI后Bcl-2、HO-1、NQO1和GCLC的mRNA相对表达量(P<0.05)。P7C3-A20对TBI后的PC12细胞有抗凋亡、减轻氧化应激的修复作用。

关键词: P7C3-A20, PC12细胞, 创伤性脑损伤, 凋亡, 氧化应激

Abstract: The study was conducted to explore the repair effect of 3, 6-Dibromo-beta-fluoro-N-(3-methoxyphenyl)-9H-carbazole-9-propanamine (P7C3-A20) on traumatic brain injury (TBI) of rat adrenal medullary pheochromoma differentiated cell line (PC12 cells). The cells were divided into control group (group A), model group (group B) and 0.03 μmol·L^-1 drug treatment group (Group C), 0.3 μmol·L^-1 drug treatment group (Group D), 3 μmol·L^-1 drug treatment group (Group E) and drug control group (Group F). TBI cell models were prepared by scribing straight lines 4 mm apart horizontally and vertically using a 10 μL gun tip. The cell viability was detected by CCK8 kit, the apoptosis and reactive oxygen (ROS) situation were observed by fluorescence microscope, and the relative mRNA expression of cysteine aspartate-specific protease-3 (Caspase-3), B-cell lymphoma/leukemia-2 (Bcl-2), Heme Oxygenase-1 (HO-1), NAD (P) H: quinone oxidoreductase 1 (NQO1) and glutamate-cysteine ligase catalytic subunit (GCLC) genes were detected by fluorescence quantitative PCR instrument. The results showed that the cell viability was extremely significantly decreased after TBI modeling (P<0.01). P7C3-A20 at concentrations of 0.03 μmol·L^-1 could significantly increase the cell viability after TBI (P<0.05). P7C3-A20 at a concentration of 0.3 μmol·L^-1 could extremely significantly increase the cell viability after TBI (P<0.01). After TBI modeling, the proportion of early and late apoptotic cells increased extremely significantly (P<0.01). P7C3-A20 at a concentration of 0.03 μmol·L^-1 could significantly reduce the proportion of early apoptotic cells after TBI (P<0.05), and extremely significantly reduce the proportion of late apoptotic cells after TBI (P<0.01), P7C3-A20 at concentrations of 0.3 and 3 μmol·L^-1 could extremely significantly reduce the proportion of early and late apoptotic cells after TBI (P<0.01). After TBI modeling, the proportion of reactive oxygen species increased extremely significantly (P<0.01). P7C3-A20 at concentrations of 0.03 and 3 μmol·L^-1 could significantly reduce the proportion of reactive oxygen species after TBI (P<0.05)， 0.3 μmol·L^-1 concentration of P7C3-A20 can significantly reduce the proportion of reactive oxygen species after TBI (P<0.01). P7C3-A20 at a concentration of 0.3 μmol·L^-1 can extremely significantly reduce the relative mRNA expression of Caspase-3 after TBI (P<0.01), and significantly increase the relative mRNA expression of Bcl-2, HO-1, NQO1 and GCLC genes expression level after TBI (P<0.05). P7C3-A20 has the repair effect of anti apoptosis and reducing oxidative stress on PC12 cells after traumatic brain injury.

Key words: P7C3-A20, PC12 cells, traumatic brain injury, apoptosis, oxidative stress

中图分类号:

S854.53

杨芷清, 张浩权, 冼润浠, 李欣然. P7C3-A20对创伤性脑损伤的PC12细胞凋亡和氧化的影响[J]. 畜牧兽医学报, 2022, 53(12): 4429-4438.

YANG Zhiqing, ZHANG Haoquan, XIAN Runxi, LI Xinran. Effects of P7C3-A20 on Apoptosis and Oxidation of PC12 Cells in Traumatic Brain Injury[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(12): 4429-4438.

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