肉豆蔻木酚素通过PI3K/Akt信号通路调节HT22细胞自噬和凋亡的作用机制  

Regulatory Mechanism of Macelignan on Autophagy and Apoptosis of HT22 Cells via PI3K/Akt Signaling Pathway

作　　者：齐正宇 邱新茹 楚新歌 李莎 延光海[1] 崔春爱[1] QI Zhengyu;QIU Xinru;CHU Xinge;LI Sha;YAN Guanghai;CUI Chun’ai(Medical College,Yanbian University,Yanji 133000,China;Songyuan Jilin Oilfield Hospital,Songyuan 138000,China)

机构地区：[1]延边大学医学院,吉林延吉133000 [2]松原吉林油田医院,吉林松原138000

出　　处：《食品科学》2025年第9期1-10,共10页Food Science

基　　金：国家自然科学基金地区科学基金项目(82060746,81660687);吉林省科技发展计划项目(YDZJ202501ZYTS235)。

摘　　要：为探究天然成分肉豆蔻木酚素对海马神经元凋亡与自噬的调控机制,利用谷氨酸诱导HT22细胞制备氧化应激细胞模型,研究肉豆蔻木酚素对HT22细胞存活率的影响。通过苏木精-伊红(hematoxylin-eosin,HE)染色、脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling,TUNEL)染色和流式细胞术评估细胞形态变化与凋亡情况;通过分子对接技术分析肉豆蔻木酚素与磷酸肌醇3-激酶(phosphatidylinositide 3-kinase,PI3K)结合情况;采用免疫印迹法分析肉豆蔻木酚素和PI3K抑制剂LY294002或自噬抑制剂3-甲基腺嘌呤(3-methyladenine,3-MA)对PI3K、蛋白激酶B(protein kinase B,Akt)的蛋白表达水平变化的影响以及凋亡与自噬相关蛋白活化半胱胺酸蛋白酶-3(Cleaved caspase-3)、微管相关蛋白轻链3β(microtubule associated protein light chain 3 beta,LC3B)、自噬相关基因5(autophagy related gene 5,Atg5)等相关蛋白表达的影响。结果显示,与氧化应激细胞模型相比,肉豆蔻木酚素可显著增强HT22细胞活性、减少凋亡细胞数量,且呈剂量依赖性,其能增强p-PI3K和p-Akt磷酸化、抑制Cleaved caspase-3表达,通过PI3K/Akt通路发挥抗凋亡作用;降低Atg5和LC3B表达、增加p62水平,有抑制自噬的作用。LY294002可阻断肉豆蔻木酚素促进PI3K和Akt磷酸化的作用,3-MA可增强其神经保护效果。综上,肉豆蔻木酚素通过促进PI3K/Akt信号通路激活、调控自噬抑制、减少谷氨酸诱导的HT22细胞凋亡从而发挥神经保护作用。To explore the regulatory mechanism of the natural compound macelignan on apoptosis and autophagy in hippocampal neurons,this study investigated the effect of macelignan on the survival rate of HT22 cell line under glutamateinduced oxidative stress.Morphological changes and apoptosis were assessed by hematoxylin-eosin(HE)staining,terminal deoxynucleotidyl transferase(TDT)-mediated dUTP nick-end labeling(TUNEL)staining and flow cytometry.The binding of macelignan with phosphatidylinositide 3-kinase(PI3K)was analyzed by molecular docking.Western blot was used to analyze the effects of macrolignan combined with the PI3K inhibitor LY294002 or the autophagy inhibitor 3-methyladenine(3-MA)on the protein expression levels of PI3K and protein kinase B(Akt),as well as apoptosis and autophagy-related proteins such as cleaved caspase-3,microtubule-associated protein light chain 3 beta(LC3B),and autophagy related gene 5(Atg5).The results demonstrated that macelignan promoted HT22 cell viability and suppressed apoptosis in a concentrationdependent manner.It could enhance the phosphorylation of p-PI3K and p-Akt,inhibit the expression of cleaved caspase-3,and exert an anti-apoptotic effect via the PI3K/Akt signaling pathway.Meanwhile,it could inhibit autophagy by reducing the expression of Atg5 and LC3B and increasing the level of p62.LY294002 could block the promoting effect of macelignan on the phosphorylation of PI3K and Akt,while 3-MA could enhance its neuroprotective effect.In conclusion,macelignan exerts a neuroprotective effect through activating the PI3K/Akt signaling pathway,inhibiting autophagy and attenuating glutamateinduced apoptosis in HT22 cells.

关 键 词：肉豆蔻木酚素 谷氨酸 神经退行性疾病 凋亡 自噬 

分 类 号：TS201.4[轻工技术与工程—食品科学]