青娥丸调控PERK/eI2αF/ATF4/CHOP通路对成骨细胞凋亡的影响  

Mechanism of Qing'E formula regulating PERK/eIF2α/ATF4/CHOP pathway and its impact on osteoblast apoptosis

作　　者：刘合亮 林煜 郑世雄 肖莉莉 LIU Heliang;LIN Yu;ZHENG Shixiong;XIAO Lili(Department of Orthopedics,Fuzhou Second General Hospital,Fuzhou,Fujian 350007,China)

机构地区：[1]福建省福州市第二总医院,福州350007

出　　处：《福建医药杂志》2025年第3期66-71,共6页Fujian Medical Journal

基　　金：福州市卫健系统科技计划项目(2021-S-wq20);福建省科学自然基金项目(2022J011318);福建省卫生健康中青年骨干人才培养项目(2020GGB046);福建省科技创新平台项目(福建省创伤骨科急救与康复临床医学研究中心,2020Y2014)。

摘　　要：目的基于PERK/eIF2α/ATF4/CHOP信号通路探讨青娥丸含药血清对氧化应激诱导内质网应激凋亡成骨细胞系的保护作用及其作用机制。方法30只雄性SD大鼠随机分为生理盐水组和青娥丸组,每组15只,分别给予生理盐水和青娥丸灌胃干预,连续7 d,腹主动脉取全血制备含药血清;培养UMR-106细胞株,CCK-8筛选青娥丸和GSK2606414最佳干预浓度,细胞分为空白组、模型组、青娥丸组和抑制剂组,进一步采用10μmol/L的H_(2)O_(2)干预细胞12 h制备内质网应激模型,CCK-8检测4组细胞的增殖活性,DCFH-DA检测细胞活性氧(ROS)含量,Real-time PCR和Western blot检测PERK、eIF2α、ATF4、CHOP mRNA和蛋白的相对表达量。结果青娥丸含药血清最佳干预浓度为10%,GSK2606414最佳干预浓度为4%,此最佳干预浓度用于后续实验。H_(2)O_(2)干预后模型组细胞增殖活性显著降低,ROS含量显著升高,PERK、eIF2α、ATF4、CHOP mRNA和蛋白的相对表达量显著上调;青娥丸组增殖活性增强,ROS含量显著降低,PERK、eIF2α、ATF4、CHOP mRNA和蛋白的相对表达量下调。结论青娥丸含药血清可有效抑制成骨细胞系的内质网应激凋亡,其作用机制可能与PERK/eIF2α/ATF4/CHOP信号通路相关。Objective To examine the protective effects and its mechanism of Qing'E formula serum on osteoblast apoptosis caused by oxidative stress-induced endoplasmic reticulum stress,focusing on the PERK/eIF2α/ATF4/CHOP signaling pathway.Methods Thirty male SD rats were randomly assigned to receive either normal saline or Qing'E formula via gavage for seven days to prepare drug-containing serum from abdominal aorta blood.UMR-106 cells were cultured,CCK-8 was selected for the optimal intervention concentration of Qing'E formula and GSK2606414.Cells were categorized into blank,model,Qing'E formula,and positive drug groups.The endoplasmic reticulum stress model was created by treating cells with 10μmol/L H_(2)O_(2) for 12 hours.Cell proliferation was assessed using CCK-8,ROS levels were dectected with DCFH-DA,and the expression levels of PERK,eIF2α,ATF4,and CHOP mRNA and protein were detected through Real-time PCR and Western blot.Results The optimal concentrations for Qing'E formula serum and GSK2606414 were found to be 10% and 4%,respectively.This optimal intervention concentration was used for subsequent experiments.Post-H_(2)O_(2) treatment,the model group showed reduced cell proliferation,increased ROS,and elevated PERK,eIF2α,ATF4,and CHOP expression.Conversely,the Qing'E formula group exhibited increased proliferation,reduced ROS,and decreased expression of the above related markers.Conclusion Qing'E formula serum effectively inhibits endoplasmic reticulum stress-induced apoptosis in osteoblasts,possibly through the PERK/eIF2α/ATF4/CHOP signaling pathway.

关 键 词：绝经后骨质疏松症 青娥丸 内质网应激 PERK/eIF2α/ATF4/CHOP信号通路 

分 类 号：R816.8[医药卫生—放射医学]