天津医药

天津医药 ›› 2022, Vol. 50 ›› Issue (6): 561-565.doi: 10.11958/20212403

• 细胞与分子生物学 •    下一篇

二苯乙烯苷、大黄素改善高糖诱导下小鼠海马神经元凋亡

陈钢   

  1. 1贵州医科大学附属医院贵州省产前诊断中心（邮编550004）；2贵州医科大学医学检验学院；3贵州医科大学公共卫生学院； 4贵州医科大学附属医院临床检验中心；5贵州中医药大学
  • 收稿日期:2021-10-25 修回日期:2022-02-24 出版日期:2022-06-15 发布日期:2023-12-20
  • 通讯作者: 陈钢 E-mail:397776980@qq.com
  • 基金资助:
    国家自然科学基金地区科学基金项目（81960151，81960822）；贵州省科技计划项目（黔科合基础-ZK［2021］366号） 

Tetrahydroxystilbene glucoside and emodin improves the hippocampal neuronal apoptosis in mice induced by high glucose

CHEN Gang1, 2, XU Yongjie1, 3, HUANG Changyudong1, 2, LIN Hairong1, 2, YANG Tingting1, 2, ZHU Liying4, #br# LI Xing5, PAN Wei1, 3△#br#   

  1. 1 The Affiliated Hospital of Guizhou Medical University, Guizhou Prenatal Diagnosis Center, Guiyang 550004, China; 
    2 School of Clinical Laboratory Science, Guizhou Medical University; 3 School of Public Health, Guizhou Medical 
    University; 4 Clinical Laboratory Center, the Affiliated Hospital of Guizhou Medical University; 
    5 Guizhou University of Traditional Chinese Medicine
  • Received:2021-10-25 Revised:2022-02-24 Published:2022-06-15 Online:2023-12-20

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摘要： 摘要：目的　探讨何首乌主要成分二苯乙烯苷（TSG）、大黄素（Emodin）对高糖诱导下海马神经元HT-22细胞凋亡的影响。方法　将小鼠海马神经元HT-22细胞分为4组：对照培养基组（NC组，葡萄糖浓度25 mmol/L）、高糖培养基组（HG组，葡萄糖浓度55 mmol/L）、HG+TSG组和HG+Emodin组。HG+TSG组和HG+Emodin组分别用TSG和Emodin以50、100、200 μmol/L的浓度作用细胞12、16、20、24、48 h，CCK-8法检测细胞存活率，确定细胞的最佳作用浓度和时间。流式细胞术检测细胞凋亡率；通过酶联免疫吸附试验（ELISA）检测4组细胞组蛋白乙酰化转移酶（HAT）和组蛋白去乙酰化酶（HDAC）活性；蛋白免疫印迹（Western blot）检测凋亡相关蛋白B淋巴细胞瘤-2（Bcl-2）、Bcl-2相关X蛋白（Bax）、半胱氨酸天冬氨酸蛋白酶3（Caspase-3）的表达。结果　TSG以200 μmol/L、48 h，Emodin以100 μmol/L、48 h作用细胞为最适条件。流式细胞术结果显示，TSG和Emodin干预细胞48 h后，细胞凋亡率显著降低。ELISA结果显示，HG+TSG组的HAT活性显著下调，HDAC活性上调；HG+Emodin组的HAT和HDAC活性显著下调。Western blot结果显示，TSG和Emodin作用细胞48 h后，Bax和Caspase-3的表达显著下调，Bcl-2的表达显著上调。结论　TSG和Emodin可能通过调节HAT和HDAC活性，从而改善在高糖诱导下的海马神经元细胞凋亡。

关键词: 海马, 神经元, 大黄素, 细胞凋亡, 组蛋白脱乙酰基酶类, 何首乌, bcl-2相关X蛋白质, 原癌基因蛋白质c-bcl-2, 半胱氨酸天冬氨酸蛋白酶3, 糖尿病脑病

Abstract: Abstract: Objective　To explore the effect of 2,3,5,4'-tetrahydroxystilbene-2-O-β-D- glucoside (TSG) and Emodin, the main components of polygonum multiflorum, on the apoptosis of hippocampal neuron HT-22 cells induced by high glucose. Methods　The mouse hippocampal neuron HT-22 cells were divided into 4 groups: the control medium group (NC group, glucose concentration 25 mmol/L), the high-glucose medium group (HG group, glucose concentration 55 mmol/L), the HG+TSG group and the HG+Emodin group. The HG+TSG group and the HG+Emodin group were treated with TSG and Emodin at the concentrations of 50, 100 and 200 μmol/L for 12, 16, 20, 24 and 48 h, respectively. The cell viability was detected by CCK-8 method after treatment. Histone acetyltransferase (HAT) and histone deacetylase (HDAC) enzyme activities were detected by enzyme-linked immunosorbent assay (ELISA). Flow cytometry was used to detect the apoptosis rate. Western blotting was used to detect the expression of apoptosis-related proteins: B-lymphoma-2 (Bcl-2), Bcl-2-related X protein (Bax) and Caspase-3. Results　The 200 μmol/L of TSG for 48 h, and 100 μmol/L of Emodin for 48 h were the optimal condition for culturing cells after testing. Flow cytometry results showed that TSG and Emodin significantly down-regulated cell apoptosis rate after 48 h of treatment. ELISA results showed that the activity of HAT was significantly down-regulated, HDAC activity was up-regulated in the HG+TSG group. The activities of HAT and HDAC were significantly down-regulated in the HG+Emodin group. Western blot results showed that the expressions levels of Bax and Caspase-3 were significantly down-regulated, and the expression of Bcl-2 was significantly up-regulated after TSG and Emodin treated cells for 48 h. Conclusion　TSG and Emodin may regulate the activities of HAT and HDAC, thereby down-regulate the apoptosis of hippocampal neurons induced by high glucose.

Key words: hippocampus, neurons, emodin, apoptosis, histone deacetylases, fallopia multiflora, bcl-2-associated X protein, proto-oncogene proteins c-bcl-2, caspase 3, diabetes encephalopathy