[1]杨　龙,刘春明,王正飞,等.银杏叶提取物对大鼠心肌缺血再灌注损伤的保护作用及机制[J].新乡医学院学报,2021,38(11):1011-1016.[doi:10.7683/xxyxyxb.2021.11.003]
　YANG Long,LIU Chunming,WANG Zhengfei,et al.Protective role and mechnism of Ginkgo biloba extract on myocardial ischemia-reperfusion injury in rats[J].Journal of Xinxiang Medical University,2021,38(11):1011-1016.[doi:10.7683/xxyxyxb.2021.11.003]
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银杏叶提取物对大鼠心肌缺血再灌注损伤的保护作用及机制
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
38
期数:
2021年11
页码:
1011-1016
栏目:
基础研究
出版日期:
2021-11-05

文章信息/Info

Title:
Protective role and mechnism of Ginkgo biloba extract on myocardial ischemia-reperfusion injury in rats
作者:
杨　龙1; 刘春明1; 王正飞1; 崔　聪2
（1.郑州市第七人民医院心血管外科，河南　郑州　450000；2.河南省胸科医院心血管外科，河南　郑州　450000）
Author(s):
YANG Long1; LIU Chunming1; WANG Zhengfei1; CUI Cong2
(1.Department of Cardiovascular Surgery,the 7th People′s Hospital of Zhengzhou,Zhengzhou 450000,Henan Province,China2.Department of Cardiovascular Surgery,Henan Provincial Chest Hospital,Zhengzhou 450000,Henan Province,China)
关键词:
银杏叶提取物; 心肌缺血再灌注损伤; 微RNA
Keywords:
ginkgo biloba extract; myocardial ischemia-reperfusion injury; microRNA
分类号:
R285.5
DOI:
10.7683/xxyxyxb.2021.11.003
文献标志码:
A
摘要:
目的　探讨银杏叶提取物（GBE）对大鼠心肌缺血再灌注损伤（MIRI）的保护作用及其机制。方法　将48只Sprague Dawley大鼠随机分为假手术组MIRI组、低剂量GBE组（L-GBE组）和高剂量GBE组（H-GBE组），每组12只。假手术组和MIRI组大鼠给予2 mL生理盐水灌胃，L-GBE组和H-GBE组大鼠分别给予50、100 mg·kg-1的GBE溶液灌胃，连续给药7 d；MIRI组、H-GBE组和L-GBE组大鼠采用栓线法制备MIRI模型；假手术组大鼠仅开脑、暴露心脏不予冠状动脉结扎。观察大鼠心肌缺血发生前10 min（T0）、缺血后30 min（T1）、再灌注30 min（T2）、再灌注60 min（T3）、再灌注120 min（T4）时心率和T4时J点抬高值。取各组大鼠心肌组织，应用苏木精-伊红染色法检测大鼠心肌组织病理形态学变化。取MI /RI组和H-GBE组大鼠心肌梗死组织，应用Illumina Hiseq 2500测序平台进行微RNA（miRNA）测序，应用实时荧光定量聚合酶链式反应(qRT-PCR)法检测差异表达miRNAs。利用miRwalk数据库对差异表达miRNAs进行靶基因预测和功能注释，通过基因本体（GO）和京都基因组百科全书数据库（KEGG）对差异表达miRNAs进行生物学过程、细胞组分、分子功能和相关信号通路预测。结果　4组大鼠T0时心率比较差异无统计学意义（F=6.659，P>0.05）。MIRI组、L-GBE组和H-GBE组大鼠T1、T2、T3、T4时心率显著低于假手术组，J点抬高值显著大于假手术组（P<0.05）；L-GBE组和H-GBE组大鼠T1、T2、T3和T4时心率显著高于MIRI组，J点抬高值显著小于MIRI组（P<0.05）；H-GBE组大鼠T1、T2、T3和T4时心率显著高于L-GBE组，J点抬高值显著小于L-GBE组（P<0.05）。假手术组大鼠心肌结构完整，细胞间质未见明显炎症浸润和水肿，心肌细胞未发生显著的变性和坏死；MIRI组大鼠心肌组织结构紊乱，心肌细胞肿胀，出现明显的空泡样变性坏死，炎症浸润程度明显增加；与MIRI组相比，H-GBE组和L-GBE组大鼠心肌组织紊乱程度降低，炎症浸润减少，水肿程度降低，H-GBE组大鼠心肌组织损伤程度较L-GBE组更低。MIRI组和H-GBE组大鼠心肌梗死组织基因测序共鉴定出1 290个miRNAs，其中miR-144、miR-200b、miR-21、miR-34、miR-146、miR-155和miR-200c 7个miRNAs呈差异表达。测序和qRT-PCR结果均显示，H-GBE组大鼠心肌组织miR-144、miR-200b显著高于MIRI组，miR-21、miR-34、miR-155、miR-146和miR-200c显著低于MIRI组（P<0.05）。7个差异表达miRNAs主要参与叉头框O信号通路、自噬、Hippo信号通路、轴突导向、肿瘤坏死因子信号通路、神经营养蛋白、哺乳动物雷帕霉素靶蛋白等信号通路和生物学过程。结论　MIRI可造成大鼠心功能损伤，GBE通过调控关键miRNAs的表达对MIRI导致的心功能损伤起保护作用。
Abstract:
Objective　To investigate the protective role of Ginkgo biloba extract (GBE) on myocardial ischemia-reperfusion injury (MIRI) in rats and its mechanism.Methods　Forty-eight Sprague Dawley rats were randomly divided into sham operation group,MIRI group,low-dose GBE group (L-GBE group)and high-dose GBE group (H-GBE group) ,with 12 rats in each group.Rats in the sham operation group and MIRI group were given 2 mL normal saline by gavagerats in the L-GBE group and H-GBE group were given 50,100 mg·kg-1 GBE solution by gavage,respectively,continuous administration for 7 daysMIRI models of rats were established by suture method in the MIRI group,L-GBE group and H-GBE groupthe rats in the sham operation group were only opened the chest and exposed the heart without coronary artery ligation.The heart rate of rats at 10 min before myocardial ischemia (T0),30 min after ischemia (T1),30 min after reperfusion (T2),60 min after reperfusion (T3),120 min after reperfusion (T4) and the J-point elevation value at T4 were observed.The myocardial tissue of rats in each group was taken and the pathomorphological changes of myocardial tissue were detected by hematoxylin-eosin staining.The myocardial infarction tissues of rats in the MIRI group and H-GBE group were taken,and microRNA(miRNA) was sequenced by Illumina Hiseq 2500 sequencing platform,the differentially expressed miRNAs were detected by real-time fluorescence quantitative polymerase chain reaction(qRT-PCR).The miRwalk database was used for target gene prediction and functional annotation of differentially expressed miRNAs,and the biological process,cell composition,molecular function and related signal pathways of differentially expressed miRNAs were predicted through gene ontology (GO) and Kyoto genome encyclopedia database (KEGG).Results　There was no significant difference in heart rate of rats at T0 among the four groups (F=6.659,P>0.05).The heart rate of rats at T1,T2,T3 and T4 in the MIRI group,L-GBE group and H-GBE group was significantly lower than that in the sham operation group,and the J-point elevation value was significantly higher than that in the sham operation group (P<0.05)the heart rate at T1,T2,T3 and T4 in the L-GBE group and H-GBE group was significantly higher than that in MIRI group,and the J-point elevation value was significantly lower than that in the MIRI group (P<0.05)the heart rate at T1,T2,T3 and T4 in the H-GBE group was significantly higher than that in the L-GBE group,and the J-point elevation value was significantly lower than that in the L-GBE group (P<0.05).In the sham operation group,the myocardial structure of rats was intact,there was no obvious inflammatory infiltration and edema in the intercellular matrix,and there was no significant degeneration and necrosis of cardiomyocytesin the MIRI group,myocardial tissue structure of rats was disordered,myocardial cells swelled,obvious vacuolar degeneration and necrosis appeared,and the degree of inflammatory infiltration increased significantlycompared with the MIRI group,the degrees of myocardial tissue disorder,inflammatory infiltration and edema were decreased in the L-GBE group and H-GBE groupthe degree of myocardial tissue injury in the H-GBE group was significantly lower than that in the L-GBE group.A total of 1 290 miRNAs in myocardial infarction tissues of rats were identified by gene sequencing in MIRI group and H-GBE group,in which the miR-144,miR-200b,miR-21,miR-34,miR-146,miR-155 and miR-200c were differentially expressed.The sequencing and qRT-PCR results showed that the levels of miR-144 and miR-200b in myocardial tissue of rats in the H-GBE group were significantly higher than those in the MIRI group,and the levels of miR-21,miR-34,miR-155,miR-146 and miR-200c were significantly lower than those in the MIRI group(P<0.05).The seven differentially expressed miRNAs were mainly involved in forkhead box O signal pathway,autophagy,Hippo signal pathway,axon guidance,tumor necrosis factor signal pathway,neurotrophic protein,mammalian rapamycin target protein and other signal pathways and biological processes.Conclusion　MIRI can cause cardiac function injury of rats,GBE can protect cardiac function injury caused by MIRI by regulating the expression of key miRNAs.

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更新日期/Last Update: 2021-11-05