羟基红花黄色素A抑制糖氧剥夺/复糖复氧处理后神经元焦亡的作用  

Inhibitory effect of hydroxy safflower yellow A on neuronal pyroptosis after glucose-oxygen deprivation/reglucose-reoxygenation treatment

作　　者：王泽乾 段彦哲 吴艺舸 马东 黄建军 闫玉清 宋丽娟 Wang Zeqian;Duan Yanzhe;Wu Yige;Ma Dong;Huang Jianjun;Yan Yuqing;Song Lijuan(The Key Research Laboratory of Benefiting Qi for Acting Blood Circulation Method to Treat Multiple Sclerosis of State Administration of Traditional Chinese Medicine/Research Center of Neurobiology,Shanxi University of Chinese Medicine,Jinzhong 030619,Shanxi Province,China;Department of Neurosurgery/The Key Laboratory of Prevention and Treatment of Neurological Disease of Shanxi Provincial Health Commission,Sinopharm Tongmei General Hospital,Datong 037003,Shanxi Province,China;Institute of Brain Science,Shanxi Datong University,Datong 037009,Shanxi Province,China;The Key Laboratory of Cellular Physiology,Ministry of Education,Shanxi Medical University,Taiyuan 030001,Shanxi Province,China)

机构地区：[1]山西中医药大学国家中医药管理局多发性硬化益气活血重点研究室/神经生物学研究中心,山西省晋中市030619 [2]国药同煤总医院神经外科/山西省神经疾病防治研究委级重点实验室,山西省大同市037003 [3]山西大同大学脑科学研究所,山西省大同市037009 [4]山西医科大学细胞生理学省部共建教育部重点实验室,山西省太原市030001

出　　处：《中国组织工程研究》2025年第19期4044-4051,共8页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(82004028),项目负责人:宋丽娟;中国博士后科学基金面上资助项目(2020M680912),项目负责人:宋丽娟;国家中医药管理局“张仲景传承与创新专项”(GZY-KJS-2022-048-1),项目负责人:宋丽娟;山西省科技创新人才青年团队项目(202204051001028),项目负责人:宋丽娟;山西中医药大学2022年度科技创新团队项目(2022TD2010),项目负责人:宋丽娟;山西省卫健委医学科技领军团队项目(2020TD05),项目负责人:马存根(执行:宋丽娟);山西中医药大学附属医院国家区域中医医疗中心心血管专项基金项目(XGZX202115),项目负责人:宋丽娟;山西中医药大学青年科学家培育项目(2021PY-QN-09),项目负责人:宋丽娟;山西中医药大学学科建设经费(2023XKJS-02),项目负责人:马存根(执行:宋丽娟)。

摘　　要：背景:羟基红花黄色素A具有抗缺血、抗氧化、抗血栓及抗炎等作用,其是否影响糖氧剥夺/复糖复氧处理后神经元焦亡,目前尚不清楚。目的:探讨羟基红花黄色素A对神经元焦亡的干预作用及机制。方法:取对数生长期的HT22细胞,随机分为5组:正常组、模型组、羟基红花黄色素A组、Colivelin组、Colivelin+羟基红花黄色素A组。利用糖氧剥夺/复糖复氧处理HT22细胞建立神经元焦亡模型,然后给予STAT3激动剂Colivelin、羟基红花黄色素A干预。干预后JC-1探针检测线粒体膜电位变化,活性氧试剂盒检测细胞内活性氧含量,GSDMD/TUNEL染色观察细胞焦亡情况,免疫荧光检测STAT3、GSDMD蛋白表达,RT-PCR检测STAT3、NLRP3、Caspase-1 mRNA表达,Western blot检测p-STAT3、NLRP3、GSDMD、Cleaved-caspase-1、白细胞介素1β蛋白表达。结果与结论:①与正常组相比,模型组焦亡细胞数量增多,p-STAT3、NLRP3、Cleaved-caspase-1、GSDMD、白细胞介素1β蛋白表达显著升高;与模型组相比,羟基红花黄色素A组焦亡细胞数量减少,焦亡相关蛋白的表达显著降低;②与模型组相比,Colivelin组细胞焦亡加剧,线粒体膜电位降低,活性氧含量增加,STAT3、NLRP3、Caspase-1 mRNA表达升高,p-STAT3、NLRP3、GSDMD、Cleaved-caspase-1、白细胞介素1β蛋白表达升高;与Colivelin组相比,Colivelin+羟基红花黄色素A组上述指标均有好转。结果表明:羟基红花黄色素A通过STAT3信号通路抑制糖氧剥夺/复糖复氧后HT22细胞焦亡发挥神经保护作用。BACKGROUND:Hydroxy safflower yellow A has anti-ischemia,anti-oxidation,anti-thrombotic and anti-inflammatory effects.Whether it affects neuronal pyroptosis after glucose-oxygen deprivation/reglucose-reoxygenation is still unclear.OBJECTIVE:To investigate the protective effect of hydroxy safflower yellow A on neuronal pyroptosis and its mechanism.METHODS:HT22 cells in logarithmic growth phase were randomly divided into five groups:normal group,model group,hydroxy safflower yellow A group,colivelin group,and colivelin+hydroxy safflower yellow A group.HT22 cells were treated with glucose-oxygen deprivation/reglucose-reoxygenation to establish neuronal pyroptosis model,and then treated with STAT3 agonist Colivelin and hydroxy safflower yellow A.JC-1 probe was employed to assess changes in mitochondrial membrane potential.Reactive oxygen species kit was used to determine the content of reactive oxygen species in cells.GSDMD/TUNEL staining was conducted to observe cell pyroptosis.Immunofluorescence analysis was performed to detect STAT3 and GSDMD protein expression.RT-PCR was utilized for assessing mRNA expression levels of STAT3,NLRP3,and Caspase-1.Western blot assay was utilized to measure the protein expression levels of p-STAT3,NLRP3,GSDMD,Cleaved-caspase-1,and interleukin-1β.RESULTS AND CONCLUSION:(1)Compared with the normal group,the number of pyroptotic cells increased in HT22 cells in the model group along with a significant increase in protein expression levels of p-STAT3,NLRP3,Cleaved-caspase-1,GSDMD,and interleukin-1β.Compared with the model group,the number of pyroptotic cells reduced,and the expression of pyroptosis-related proteins significantly decreased in the hydroxy safflower yellow A group.(2)In comparison with the model group,pyroptosis worsened in the colivelin group where mitochondrial membrane potential decreased along with elevated reactive oxygen species content and increased mRNA expression levels of STAT3,NLRP3,and Caspase-1,as well as increased protein expression levels of p-STAT3,NLRP3,GS

关 键 词：HT22细胞 细胞焦亡 羟基红花黄色素A 神经元 糖氧剥夺 复糖复氧 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R741.02