|本期目录/Table of Contents|

[1]禹博,祁琳,金鑫,等.芹菜素对化学法氧糖剥夺/复氧复糖损伤大鼠心肌细胞保护作用研究[J].天津医科大学学报,2023,29(03):258-264.
　YU Bo,QI Lin,JIN Xin,et al.The protective effect of apigenin on cardiomyocytes of rats injured by chemical oxygen-glucose deprivation/restoration[J].Journal of Tianjin Medical University,2023,29(03):258-264.
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芹菜素对化学法氧糖剥夺/复氧复糖损伤大鼠心肌细胞保护作用研究(PDF)
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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:
29卷
期数:
2023年03期
页码:
258-264
栏目:
基础医学
出版日期:
2023-05-20

文章信息/Info

Title:
The protective effect of apigenin on cardiomyocytes of rats injured by chemical oxygen-glucose deprivation/restoration
文章编号:
1006－8147（2023）03-0258-07
作者:
禹博1; 祁琳2; 金鑫3; 曹清文4; 田晨4; 李佳颖4; 王越4; 周建妹5; 陈康寅1
（1.天津医科大学第二医院心脏科，天津市心血管病离子与分子机能重点实验室，天津心脏病学研究所，天津300211；2.武警特色医学中心药剂科，天津300162；3.武警后勤学院军事药学教研室，天津300309；4.天津中医药大学中西医结合学院，天津301617；5.浙江医院干部保健科，杭州310013）
Author(s):
YU Bo1; QI Lin2; JIN Xin3; CAO Qing-wen4; TIAN Chen4; LI Jia-ying4; WANG Yue4; ZHOU Jian-mei5; CHEN Kang-yin1
关键词:
芹菜素; 氧糖剥夺/复氧复糖; 氯化钴; 心肌细胞
Keywords:
apigenin; oxygen-glucose deprivation/restoration; cobalt chloride; cardiomyocytes
分类号:
R541
DOI:
-
文献标志码:
A
摘要:
目的：研究芹菜素（API）对化学法氧糖剥夺/复氧复糖（OGD/R）大鼠H9c2心肌细胞损伤的保护作用及分子机制。方法：首先给予H9c2细胞不同浓度的氯化钴（CoCl2）或1%O2处理24 h，通过测定细胞活力和低氧诱导因子-1α（HIF-1α）的表达水平，确定模拟1%O2的等效CoCl2浓度；然后给予H9c2细胞CoCl2或1%O2处理合并无糖培养16 h，再正常培养（复氧复糖）4 h，测定细胞活力和HIF-1α的表达水平，确定OGD/R模型等效CoCl2浓度。MTT法测定不同浓度API对OGD/R大鼠H9c2细胞活力的影响；ELISA法检测细胞Caspase-3含量；超氧化物歧化酶（SOD）和乳酸脱氢酶（LDH）试剂盒测定细胞SOD和LDH含量；采用Western 印迹方法检测细胞Bcl-2、Bax、SIRT1、p62和LC3Ⅱ/Ⅰ表达水平。结果：筛选1%O2的等效CoCl2浓度结果显示，与对照组相比，CoCl2（0.8 mmol/L）组细胞活力明显降低（P<0.05），HIF-1α表达明显上调（P<0.05）且与1%O2组无显著差异（P>0.05），因此选取CoCl2（0.8 mmol/L）用于OGD/R模型。与对照组相比，OGD/R组的细胞活力及SOD含量明显下降（P<0.05），LDH漏出量和Caspase-3含量明显提高（P<0.05），SIRT1、LC3Ⅱ/Ⅰ及Bcl-2表达量均明显下降（均P<0.05），Bax及p62的表达量明显增加（P<0.05）；与OGD/R组相比，API（0.01～10 μmol/L）组细胞活力有显著提高（P<0.05）；10 μmmol/L API能显著提高细胞SOD含量（P<0.05），显著降低LDH漏出量和Caspase-3含量（均P<0.05），明显提高SIRT1、LC3Ⅱ/Ⅰ及Bcl-2的表达量（均P<0.05），降低Bax及p62的表达量（均P<0.05）。结论：0.8 mmol/L CoCl2合并剥糖16 h后恢复4 h可构建稳定的H9c2细胞OGD/R损伤模型，API可通过调节SIRT1增强自噬以抵抗H9c2细胞OGD/R损伤和凋亡。
Abstract:
Objective：To investigate the protective effect and molecular mechanism of apigenin（API） on H9c2 cardiomyocytes of rats injured by chemical glucose-oxygen deprivation/restoration（OGD/R）. Methods：Firstly，H9c2 cells were treated with different concentrations of cobalt chloride（CoCl2） or 1%O2 for 24 h. The cell viability and the expression level of hypoxia-inducible factor-1α（HIF-1α） were measured to determine the equivalent CoCl2 concentration to simulate 1%O2. Then H9c2 cells were treated with CoCl2 or 1%O2 and cultured without glucose for 16 h，and then cultured normally（restoration）for 4 h. Cell viability and HIF-1α expression level were measured to determine the equivalent CoCl2 concentration of the OGD/R model. MTT assay was used to determine the effects of different concentrations of API on the viability of H9c2 cells injured by OGD/R. The content of Caspase-3 was detected by ELISA. Superoxide dismutase（SOD） and lactate dehydrogenase（LDH） kits were used to determine the contents of SOD and LDH. The expression levels of Bcl-2，Bax，SIRT1，p62 and LC3Ⅱ/Ⅰ were detected by Western blotting. Results：The results of screening the equivalent CoCl2 concentration to simulate 1%O2 showed that compared with the control group，the cell viability of CoCl2（0.8 mmol/L） group was significantly decreased and the HIF-1α expression was significantly up-regulated（P<0.05）；and there was no significant difference between 1% O2 and the above results. Therefore，CoCl2（0.8 mmol/L） was selected for OGD/R model. Compared with the control group，the cell viability and SOD content of the OGD/R group were significantly decreased（P<0.05）；LDH leakage and Caspase-3 content in the OGD/R group were significantly increased（P<0.05）；the expressions of SIRT1，LC3Ⅱ/Ⅰ and Bcl-2 in OGD/R group were significantly decreased（all P<0.05），the expressions of Bax and p62 were significantly increased（both P<0.05）. Compared with the OGD/R group，the API（0.01-10 μmol/L） group significantly increased cell viability（P<0.05），10 μmol/L API could significantly increase the SOD content of cells（P<0.05），decrease the LDH leakage and Caspase-3 content（P<0.05），increase the expression of SIRT1，LC3Ⅱ/Ⅰ and Bcl-2（all P<0.05） and decrease the expression of Bax and p62（both P<0.05）.Conclusion：The stable OGD/R injury model of H9c2 cells can be established by 0.8 mmol/L CoCl2 combined with depriving glucose for 16 h and oxygen-glucose recovery for 4 h. API can protect H9c2 cells from OGD/R injury and apoptosis by regulating SIRT1 to enhance autophagy.

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备注/Memo

备注/Memo:
基金项目 浙江省医药卫生科技计划项目（2021KY013）；天津市医学重点学科（专科）建设资助项目（TJYXZDXK-029A）；天津中医药大学研究生创新基金（ZXYCXLX202119）；天津中医药大学研究生创新基金（ZXYCXLX202018）；武警部队创新研究项目（ZZKY20222419）；武警后勤学院基础研究项目（WHJ202007）
作者简介 禹博（1995-），男，硕士在读，研究方向：心血管内科介入治疗；通信作者：陈康寅，E-mail：chenkangyin@vip.126.com；周建妹，E-mail：zhou1306928@163.com。
更新日期/Last Update: 1900-01-01