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谢薇, 彭俊, 宋厚盼, 欧晨, 彭清华.基于PERK/ATF4/CHOP信号通路研究滋阴明目方含药血清对衣霉素诱导的ARPE-19细胞的作用机制[J].湖南中医药大学学报英文版,2024,44(5):785-790.[Click to copy
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| 基于PERK/ATF4/CHOP信号通路研究滋阴明目方含药血清对衣霉素诱导的ARPE-19细胞的作用机制 |
| 谢薇,彭俊,宋厚盼,欧晨,彭清华 |
| (湖南中医药大学, 湖南 长沙 410208;湖南中医药大学第一附属医院, 湖南 长沙 410007) |
| 摘要: |
| 目的 研究滋阴明目方含药血清对衣霉素诱导ARPE-19细胞的影响及其可能机制。方法 构建细胞内质网应激损伤模型，将ARPE-19细胞分为空白组、模型组、空白血清组、滋阴明目方含药血清组、牛磺熊去氧胆酸组。对细胞进行形态观察，CCK-8检测细胞存活率，TUNEL法检测细胞凋亡，Western blot检测细胞蛋白激酶样内质网激酶（PERK）、活化转录因子4（ATF4）、C/EBP同源蛋白（CHOP）蛋白的表达。结果 选用浓度50 μmol/L衣霉素干预ARPE-19细胞造模。观察细胞形态发现，滋阴明目方含药血清组和牛磺熊去氧胆酸组ARPE-19细胞较模型组细胞数量增多，生长较均匀，漂浮的死亡ARPE-19细胞及碎片减少。与空白组相比，模型组和空白血清组的细胞存活率下降（P＜0.01），凋亡率明显上升（P＜0.01），PERK、ATF4、CHOP蛋白表达上调（P＜0.01）。与模型组相比，滋阴明目方含药血清组细胞存活率上升（P＜0.01），凋亡率明显下降（P＜0.01），PERK、ATF4、CHOP蛋白表达下调（P＜0.01）。结论 滋阴明目方含药血清可以减少ARPE-19细胞内质网应激损伤模型的凋亡，其分子机制与调控PERK-ATF4-CHOP信号通路有关。 |
| 关键词: 滋阴明目方 ARPE-19细胞 衣霉素 内质网应激损伤模型 PERK/ATF4/CHOP信号通路 |
| DOI：10.3969/j.issn.1674－070X.2024.05.010 |
| Received:October 11, 2023 |
| 基金项目:国家自然科学基金面上项目（82074196，82004427）；湖南省自然科学基金项目（2023JJ40474）；湖南省教育厅科学研究项目（23B0347）；中医药防治五官科疾病湖南省重点实验室建设项目（2017TP1018）。 |
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| Mechanism of action of Ziyin Mingmu Formula medicated serum on tunicamycin-induced ARPE-19 cells based on PERK/ATF4/CHOP signaling pathway |
| XIE Wei, PENG Jun, SONG Houpan, OU Chen, PENG Qinghua |
| (Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;The First Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China) |
| Abstract: |
| Objective To investigate the effects of Ziyin Mingmu Formula (ZYMMF) medicated serum on tunicamycin (TM)-induced ARPE-19 cells and its potential mechanism. Methods The endoplasmic reticulum stress injury model was constructed and ARPE-19 cells were divided into blank control, model, blank control serum, ZYMMF medicated serum, and taurodeoxycholic acid groups. Cell morphology was observed, cell survival rate was determined by CCK-8, cell apoptosis was tested by TUNEL, and protein expressions of PERK kinase (PERK), activating transcription factor 4 (ATF4), and transcription factor CHOP (CHOP) were measured by Western blot. Results ARPE-19 cells were treated with 50 μmol/L tunicamycin for modeling. Compared with the model group, the number of ARPE-19 cells in ZYMMF medicated serum and taurodeoxycholic acid groups increased with more uniform growth and reduced floating dead ARPE-19 cells and fragments. Compared with blank control group, the cell survival rate in model group and blank control serum group decreased (P<0.01), the apoptosis rate increased significantly (P<0.01), and the protein expressions of PERK, ATF4, and CHOP were significantly up-regulated (P<0.01). Compared with the model group, the survival rate of ZYMMF medicated serum group increased (P<0.01), the apoptosis rate decreased significantly (P<0.01), and the protein expressions of PERK, ATF4, and CHOP were significantly down-regulated (P<0.01). Conclusion ZYMMF medicated serum can reduce the apoptosis of endoplasmic reticulum stress injury model of ARPE-19 cells, and its molecular mechanism is related to the regulation of PERK/ATF4/CHOP signaling pathway. |
| Key words: Ziyin Mingmu Formula ARPE-19 cells tunicamycin endoplasmic reticulum stress injure model PERK/ATF4/CHOP signaling pathway |
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