中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (25): 6566-6574.doi: 10.12307/2026.463

• 组织构建细胞学实验 cytology experiments in tissue construction • 上一篇    下一篇

血糖波动对小鼠海马神经元细胞HT-22凋亡的作用与机制

陈  迪1，2，徐梦玲1，2，饶彬阐1，2，朱丽英1，3，李  兴4，许永劼2，潘  卫1，2   

  1. 1贵州医科大学医学检验学院，贵州省贵阳市   550004；贵州医科大学附属医院，2贵州省产前诊断中心，3临床检验中心，贵州省贵阳市   550004；4贵州中医药大学基础医学院，贵州省贵阳市   550025
  • 收稿日期:2025-09-15 修回日期:2026-02-12 出版日期:2026-09-08 发布日期:2026-04-22
  • 通讯作者: 潘卫，博士，教授，贵州医科大学医学检验学院，贵州省贵阳市 550004；贵州医科大学附属医院，贵州省产前诊断中心，贵州省贵阳市 550004 共同通讯作者：许永劼，博士，贵州医科大学附属医院，贵州省产前诊断中心，贵州省贵阳市 550004
  • 作者简介:陈迪，女，1998年生，贵州省毕节市人，汉族，贵州医科大学在读硕士，主要从事糖尿病并发症发病机制研究。
  • 基金资助:
    国家自然科学基金地区科学基金项目(82260165)，项目负责人：潘卫；国家自然科学基金青年科学基金项目(82300920)，项目负责人：许永劼；贵州省科技计划项目(黔科合基础-ZK[2024]一般199)，项目负责人：许永劼；贵州省卫生健康委科学技术基金项目(gzwkj2024-081)，项目负责人：许永劼；贵州医科大学附属医院国家自然科学基金地区基金培育计划项目(gyfyhsfc-2022-37)，项目负责人：朱丽英；国家自然科学基金项目(82560169)，项目负责人：朱丽英

Effects and mechanisms of glycemic variability on apoptosis in mouse hippocampal neuronal HT-22 cells

Chen Di1, 2, Xu Mengling1, 2, Rao Binchan1, 2, Zhu Liying1,3, Li Xing4, Xu Yongjie2, Pan Wei1, 2   

  1. 1School of Medical Laboratory Science, Guizhou Medical University, Guiyang 550004, Guizhou Province, China; 2Guizhou Provincial Prenatal Diagnosis Center, 3Clinical Laboratory Center, Guizhou Medical University Affiliated Hospital, Guiyang 550004, Guizhou Province, China; 4School of Basic Medical Sciences, Guizhou University of Traditional Chinese Medicine, Guiyang 550025, Guizhou Province, China
  • Received:2025-09-15 Revised:2026-02-12 Online:2026-09-08 Published:2026-04-22
  • Contact: Pan Wei, PhD, Professor, School of Medical Laboratory Science, Guizhou Medical University, Guiyang 550004, Guizhou Province, China; Guizhou Provincial Prenatal Diagnosis Center, Guizhou Medical University Affiliated Hospital, Guiyang 550004, Guizhou Province, China Co-corresponding author: Xu Yongjie, PhD, Guizhou Provincial Prenatal Diagnosis Center, Guizhou Medical University Affiliated Hospital, Guiyang 550004, Guizhou Province, China
  • About author:Chen Di, MS candidate, School of Medical Laboratory Science, Guizhou Medical University, Guiyang 550004, Guizhou Province, China; Guizhou Provincial Prenatal Diagnosis Center, Guizhou Medical University Affiliated Hospital, Guiyang 550004, Guizhou Province, China
  • Supported by:
    the Regional Science Foundation Project of the National Natural Science Foundation of China, No. 82260165 (to PW); the National Natural Science Foundation of China (Youth Program), No. 82300920 (to XYJ); the Guizhou Provincial Science and Technology Plan Project, No. ZK[2024] General 199 (to XYJ); the Science and Technology Foundation of Guizhou Provincial Health Commission, No. gzwkj2024-081 (to XYJ); the National Natural Science Foundation Regional Fund Cultivation Program of the Affiliated Hospital of Guizhou Medical University, No. gyfyhsfc-2022-37 (to ZLY); the National Natural Science Foundation of China, No. 82560169 (to ZLY) 

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摘要：



文题释义：
细胞凋亡：也被称为程序性细胞死亡，是一种由基因控制的主动有序的细胞死亡方式，在多细胞生物的生长发育、组织稳态维持以及疾病发生发展等过程中发挥着至关重要的作用。  
沉默信息调节因子1：属于Ⅲ类组蛋白去乙酰化酶家族成员，能通过对多种非组蛋白及组蛋白赖氨酸残基进行去乙酰化修饰调节基因表达。沉默信息调节因子1能感受细胞中的能量水平，具有延缓细胞衰老、帮助细胞抵御外界应激和改善代谢的功能。

背景：前期研究证实，持续高糖环境引发的“代谢记忆”效应可通过调控组蛋白乙酰化酶活性显著加剧小鼠海马神经元细胞HT-22的
损伤。
目的：探讨血糖波动、持续高糖对小鼠海马神经元细胞HT-22凋亡以及组蛋白去乙酰化酶4、沉默信息调节因子1表达的影响。
方法：第6代HT-22细胞贴壁后分3组培养：对照组加入25 mmol/L葡萄糖，培养3 d或5 d；高糖组加入55 mmol/L葡萄糖，培养3 d或
5 d；血糖波动组加入25 mmol/L葡萄糖培养12 h后更换为55 mmol/L葡萄糖培养12 h，持续波动3 d或5 d。培养3 d，光学显微镜下观察细胞形态；培养5 d，流式细胞术检测细胞凋亡；培养3，4，5 d后，CCK-8法检测细胞存活率；培养3，5 d后，2，7-二氯荧光素二乙酸酯荧光探针检测细胞中活性氧水平，ELISA法检测细胞上清中组蛋白去乙酰化酶含量，Western blot检测细胞中Bax、Bcl-2、Caspase-3、Cleaved Caspase-3、沉默信息调节因子1和组蛋白去乙酰化酶4蛋白表达，RT-qPCR检测细胞中Bax、Bcl-2、Caspase-3、沉默信息调节因子1和组蛋白去乙酰化酶4 mRNA表达。
结果与结论：①光学显微镜下可见对照组细胞生长状态良好，交织成致密网状，突触间相互连接；高糖组、血糖波动组细胞生长受抑制，细胞间突触连接减少；高糖组细胞凋亡率高于对照组、血糖波动组(P < 0.05)，血糖波动组细胞凋亡率高于对照组(P < 0.05)。②培养3，4，5 d，高糖组细胞存活率低于对照组、血糖波动组(P < 0.05)，血糖波动组细胞存活率低于对照组(P < 0.05)。③培养3，5 d后，高糖组细胞中活性氧水平高于对照组、血糖波动组(P < 0.05)，血糖波动组细胞中活性氧水平高于对照组(P < 0.05)，高糖组、血糖波动组细胞上清中组蛋白去乙酰化酶含量高于对照组(P < 0.05)。④培养3，5 d后，高糖组、血糖波动组细胞中组蛋白去乙酰化酶4、Bax、Caspase-3的蛋白与mRNA表达均高于对照组(P < 0.05)，沉默信息调节因子1、Bcl-2的蛋白与mRNA表达均低于对照组(P < 0.05)，Cleaved Caspase-3蛋白表达高于对照组(P < 0.05)。⑤结果表明，血糖波动可能通过上调组蛋白去乙酰化酶4表达、下调沉默信息调节因子1表达诱导HT-22细胞凋亡。

https://orcid.org/0009-0002-8017-7625 (陈迪) 


中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 血糖波动, 血糖波动神经元模型的建立, 高糖, 细胞凋亡, 组蛋白去乙酰化酶, 小鼠海马神经元细胞, 沉默信息调节因子

Abstract: BACKGROUND: Previous studies have confirmed that the “metabolic memory” effect induced by a sustained high-glucose environment can significantly exacerbate damage in mouse hippocampal neuronal cell lines HT-22 by regulating histone acetylase activity.
OBJECTIVE: To investigate the effects of glycemic variability and sustained high glucose on apoptosis and the expression of histone deacetylase 4 and silent information regulator 1 in mouse hippocampal neuronal cell lines HT-22.
METHODS: The 6th generation of HT-22 cells were adherent and divided into three groups for culture: a control group was cultured with 25 mmol/L glucose for 3 or 5 days; a high glucose group was cultured with 55 mmol/L glucose for 3 or 5 days; a glycemic variability group was cultured with 25 mmol/L glucose for 12 hours and then switched to 55 mmol/L glucose for 12 hours, with this fluctuation lasting for 3 or 5 days. After 3 days of culture, cell morphology was observed under an optical microscope. After 5 days of culture, cell apoptosis was detected by flow cytometry. After 3, 4, and 5 days of culture, cell viability was detected by cell counting kit-8 assay. After 3 and 5 days of culture, reactive oxygen species levels in cells were detected using a DCFH-DA fluorescent probe, histone deacetylase content in the cell supernatant was detected by ELISA, protein expression of Bax, Bcl-2, Caspase-3, Cleaved Caspase-3, silent information regulator 1, and histone deacetylase 4 in cells was detected by western blot, and mRNA expression of Bax, Bcl-2, Caspase-3, silent information regulator 1, and histone deacetylase 4 in cells was detected by RT-qPCR.
RESULTS AND CONCLUSION: (1) Under an optical microscope, cells in the control group showed good growth, forming a dense network with interconnected synapses; cell growth was inhibited in the high glucose and glycemic variability groups, with reduced synaptic connections between cells. The apoptosis rate in the high glucose group was higher than that in the control and glycemic variability groups (P < 0.05), and the apoptosis rate in the glycemic variability group was higher than that in the control group (P < 0.05). (2) On days 3, 4, and 5 of culture, cell viability in the high glucose group was lower than that in the control and glycemic variability groups (P < 0.05), and cell viability in the glycemic variability group was lower than that in the control group (P < 0.05). (3) After 3 and 5 days of culture, reactive oxygen species levels in the high glucose group were higher than those in the control and glycemic variability groups (P < 0.05), reactive oxygen species levels in the glycemic variability group were higher than those in the control group (P < 0.05), and histone deacetylase content in the cell supernatant of the high glucose and glycemic variability groups was higher than that in the control group (P < 0.05). (4) After 3 and 5 days of culture, the protein and mRNA expression of histone deacetylase 4, Bax, and Caspase-3 in the high glucose and glycemic variability groups were higher than those in the control group (P < 0.05), while the protein and mRNA expression of silent information regulator 1 and Bcl-2 were lower than those in the control group (P < 0.05), and Cleaved Caspase-3 protein expression was higher than that in the control group (P < 0.05). Overall, these findings indicate that glycemic variability may induce HT-22 cell apoptosis by upregulating histone deacetylase 4 expression and downregulating silent information regulator 1 expression.

Key words: glycemic variability, establishment of a neuronal model for glycemic variability, high glucose, apoptosis, histone deacetylase, mouse hippocampal neuronal cells, silent information regulator

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