文献类型：期刊文献

中文题名：基于PERK/eIF2α/ATF4/CHOP信号通路探讨四味黄芪散对高原低氧致脑损伤大鼠的保护作用及机制

英文题名：To Investigate the Protective Effect and Mechanism of Siwei Huangqi Powder on Rats with Highaltitude Hypoxia-induced Brain Injury Based on PERK/eIF2α/ATF4/CHOP Signaling Pathway

作者：李从艺[1];曹旺杰[1];苏韫[1];龚红霞[1];刘永琦[1];和建政[1];郭家旺[1];张鑫珏[1]

第一作者：李从艺

机构：[1]甘肃中医药大学基础医学院,甘肃省高校重大疾病分子医学与中医药防治研究省级重点实验室,敦煌医学与转化教育部重点实验室,甘肃兰州730000

第一机构：甘肃中医药大学基础医学院（敦煌医学研究所）|甘肃中医药大学科研实验中心（甘肃省中医药标准化技术委员会秘书处）

年份：2024

卷号：35

期号：11

起止页码：1661

中文期刊名：中药新药与临床药理

外文期刊名：Traditional Chinese Drug Research and Clinical Pharmacology

收录：CSTPCD;;北大核心:【北大核心2023】；CSCD:【CSCD2023_2024】；

基金：甘肃省高等学校青年博士基金项目(2022QB-102);甘肃中医药大学引进人才科研启动基金项目(2023YJRC-06);甘肃中医药大学研究生创新创业项目。

语种：中文

中文关键词：四味黄芪散;高原低氧;脑损伤;PERK/eIF2α/ATF4/CHOP信号通路;内质网应激;大鼠

外文关键词：Siwei Huangqi Powder;high-altitude hypoxia;brain injury;PERK/eIF2α/ATF4/CHOP signaling pathway;endoplasmic reticulum stress;rats

摘要：目的基于PERK/eIF2α/ATF4/CHOP信号通路探讨四味黄芪散(藏黄芪、藏红花、川芎、沉香)对高原低氧致脑损伤大鼠的保护作用及机制。方法将SD大鼠随机分为空白组、模型组、阳性药物组(诺迪康,0.125 g·kg^(-1)·d-1)以及四味黄芪散低、中、高剂量组(3.255、6.510、13.020 g·kg^(-1)·d-1),每组10只。四味黄芪散各剂量组灌胃给药,每日1次,连续14 d;阳性药物组给予诺迪康灌胃给药,每日1次,连续给药7 d。采用实验动物低压模拟舱复制高原低氧致脑损伤大鼠模型,低氧暴露持续7 d。采用HE染色法观察大鼠脑组织病理变化;TUNEL染色法检测大鼠脑组织细胞凋亡情况;TBA法、微板法分别检测脑组织中丙二醛(MDA)、谷胱甘肽(GSH)的水平;Western Blot法检测大鼠脑组织中p-PERK/PERK、p-eIF2α/eIF2α、ATF4、CHOP蛋白表达水平;RT-qPCR法检测大鼠脑组织中PERK、eIF2α、ATF4、CHOP mRNA表达水平。结果与空白组比较,模型组大鼠海马CA1区锥体细胞水肿,排列不规则,胞质疏松淡染;海马CA1区红色荧光强度显著增强(P<0.01);脑组织中MDA含量显著升高(P<0.01),GSH活性显著降低(P<0.01);脑组织中p-PERK/PERK、p-eIF2α/eIF2α、ATF4、CHOP蛋白表达水平显著升高(P<0.01),PERK、eIF2α、ATF4、CHOP mRNA表达显著上调(P<0.01)。与模型组比较,阳性药物组及四味黄芪散高、中、低剂量组大鼠海马CA1区锥体细胞排列紧密,边界较清,细胞水肿得到改善;海马CA1区的红色荧光强度明显减弱(P<0.05);脑组织中MDA含量显著降低(P<0.01),GSH活性显著升高(P<0.05,P<0.01)。四味黄芪散高、中剂量组大鼠脑组织中p-PERK/PERK、p-eIF2α/eIF2α、ATF4、CHOP蛋白表达水平显著降低(P<0.05,P<0.01),PERK、eIF2α、ATF4、CHOP mRNA表达显著下调(P<0.05,P<0.01);四味黄芪散低剂量组大鼠脑组织中p-eIF2α/eIF2α、CHOP蛋白表达水平显著降低(P<0.05,P<0.01),eIF2α、CHOP mRNA表达显著下调(P<0.05,P<0.01)。结论四味黄芪散能改善高原低氧致脑损伤大鼠的脑组织神经元细胞水肿,增强抗氧化应激能力,减少脑组织细胞凋亡,可能与其调控PERK/eIF2α/ATF4/CHOP信号通路抑制内质网应激有关。
Objective To investigate the protective effect of Siwei Huangqi Powder(Astragalus tibeticola Podlech,Croci Stigma,Chuangxiong Rhizoma,Aquilariae Lignum Resinatum)on rats with high-altitude hypoxia-induced brain injury based on the protein kinase R-like endoplasmic reticulum kinase(PERK)/eukaryotic translation initiation factor 2α(eIF2α)/transcription activator 4(ATF4)/endoplasmic reticulum stress enhancer binding(C/EBP)homologous protein(CHOP)signaling pathway.Methods SD rats were randomly divided into blank group,model group,the high-,medium-and low-dose of Siwei Huangqi Powder groups(3.255,6.510,13.020 g·kg^(-1)·d-1)and positive drug group(Nuodikang,0.125 g·kg^(-1)·d-1),with 10 rats in each group.Each dose group of Siwei Huangqi Powder was administered by gavage,once a day for 14 days.The rats in the positive drug group were given Nuodikang by gavage,once a day for seven days.A rat model of plateau hypoxic brain injury was replicated in a low-pressure simulation chamber,hypoxic exposure for seven days.HE staining was used to observe pathological changes in rat brain tissue.TdT-mediated dUTP nick end-labeling(TUNEL)staining was used to detect the apoptosis of brain cells.The contents of malondialdehyde(MDA)and glutathione(GSH)in brain tissue were detected by TBA and microplate method,respectively.The protein expressions of p-PERK/PERK,p-eIF2α/eIF2α,ATF4 and CHOP in brain tissue were detected by Western Blot.RT-qPCR was used to detect the mRNA expression of PERK,eIF2α,ATF4 and CHOP in rat brain tissue.Results Compared with the blank group,the pyramidal cells in the hippocampal CA1 region of the model group were edema,irregularly arranged,and the cytoplasm was loose and lightly stained.The intensity of red fluorescence in hippocampal CA1 region was significantly increased(P<0.01).The content of MDA in brain tissue was significantly increased(P<0.01),and the activity of GSH was significantly decreased(P<0.01).The protein expression levels of p-PERK/PERK,p-eIF2α/eIF2α,ATF4 and CHOP in brain tissue were significantly increased(P<0.01).The mRNA expression levels of PERK,eIF2α,ATF4 and CHOP in brain tissue were significantly upregulated(P<0.01).Compared with the model group,the pyramidal cells in the hippocampal CA1 region of the rats in positive drug group and Siwei Huangqi Powder groups were closely arranged,the boundary was clear,and the cell edema was improved.The intensity of red fluorescence in hippocampal CA1 region was significantly decreased(P<0.05).The content of MDA in brain tissue was significantly decreased(P<0.01),and the activity of GSH was significantly increased(P<0.05,P<0.01).The protein expressions of p-PERK/PERK,p-eIF2α/eIF2α,ATF4 and CHOP in the brain tissue of the high-and medium-dose of Siwei Huangqi Powder groups were significantly decreased(P<0.05,P<0.01),while the mRNA expression levels of PERK,eIF2α,ATF4 and CHOP were significantly down-regulated(P<0.05,P<0.01).The protein expressions of p-eIF2α/eIF2αand CHOP in the brain tissue of the low-dose Siwei Huangqi Powder group were significantly decreased(P<0.05,P<0.01),while the mRNA expression levels of eIF2αand CHOP were significantly down-regulated(P<0.05,P<0.01).Conclusions Siwei Huangqi Powder can reduce the edema of neuronal cells in brain tissue,increase the ability of anti-oxidative stress,and reduce the apoptosis of brain neuronal cells in rats with high-altitude hypoxia-induced brain injury,possibly by regulating the PERK/eIF2α/ATF4/CHOP signaling pathway to inhibit endoplasmic reticulum stress.