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  

Mechanism of action of Ziyin Mingmu Formula medicated serum on tunicamycin-induced ARPE-19 cells based on PERK/ATF4/CHOP signaling pathway

作　　者：谢薇 彭俊[1,2] 宋厚盼[1] 欧晨 彭清华[1,2] 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)

机构地区：[1]湖南中医药大学,湖南长沙410208 [2]湖南中医药大学第一附属医院,湖南长沙410007

出　　处：《湖南中医药大学学报》2024年第5期785-790,共6页Journal of Hunan University of Chinese Medicine

基　　金：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)。

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50μmol/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。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 50μmol/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.

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 

分 类 号：R285.5[医药卫生—中药学]