[1]李鹏盈,卢成志△,周小波,等.银杏叶提取物EGB761对缺氧心肌细胞凋亡的保护作用及分子机制研究[J].陕西中医,2019,(1):3-8.
　LI Pengying,LU Chengzhi,ZHOU Xiaobo,et al.The effect of Ginkgo biloba extract 761 on the apoptosis related genes of myocardial cells induced by hypoxia and it’s molecular mechanism[J].,2019,(1):3-8.
点击复制

银杏叶提取物EGB761对缺氧心肌细胞凋亡的保护作用及分子机制研究
分享到：

《陕西中医》[ISSN:1000-7369/CN:61-1281/TN]

卷:
期数:
2019年1期
页码:
3-8
栏目:
基础研究
出版日期:
2019-01-05

文章信息/Info

Title:
The effect of Ginkgo biloba extract 761 on the apoptosis related genes of myocardial cells induced by hypoxia and it’s molecular mechanism
文章编号:
DOI:10.3969/j.issn.10007369.2019.01.001
作者:
李鹏盈1; 卢成志2△; 周小波1; 王桂贤1
1.天津市东丽区东丽医院（天津 300300）;2.天津市第一中心医院心内科（天津 300192）
Author(s):
LI Pengying;  LU Chengzhi; ZHOU Xiaobo; et al.
Department of Emergency, Dongli Hospital of Dongli District in Tianjin (Tianjin 300300)
关键词:
银杏叶; EGB761; 凋亡; 心肌缺氧; PPARγ; NFκBp65; 大鼠
Keywords:
Key words Ginkgo; EGB761; Apoptosis; Myocardial anoxia; PPARγ; NFκBp65; Rats
分类号:
R542.2
文献标志码:
A
摘要:
摘 要 目的：研究银杏叶提取物EGB761对缺氧原代乳鼠心肌细胞凋亡相关基因的影响及分子机制。方法：大鼠乳鼠1 d提取心肌原代细胞培养，分对照组、缺氧组、缺氧+EGB761 5μg/ml组、缺氧+EGB761 10μg/ml组、缺氧+EGB761 20μg/ml组，MTT法检测细胞存活率、ELISA法检测细胞上清LDH及细胞匀浆中SOD的活性变化、Hoechst染色检测细胞凋亡情况、PCR及Westernblot法检测PPARγ、NFκBp65、IκB、Bax、Bcl2、Caspase3 mRNA及蛋白表达。结果：和对照组相比，缺氧6 h时细胞存活率明显下降，且随着加入EGB761浓度增加，细胞存活率逐渐升高。EGB761（10μg/ml和20μg/ml浓度）可有效降低（P<0.05）缺氧造成的LDH漏出和凋亡增加，不同浓度EGB761均可升高细胞内SOD活性（P<0.05）。缺氧造成PPARγ、Ikb、抗凋亡因子Bcl2表达下降；NFκBp65基因、促凋亡因子Bax、Caspase3表达上升，不同浓度的EGB761可逆转缺氧引起的相关因子表达变化（P<0.05）。结论:银杏叶提取物EGB761可调控PPARγ、NFκBp65及凋亡关键基因的表达，并逆转缺氧引起的心肌细胞凋亡增加、存活率下降等损伤效应。
Abstract:
Abstract Objective: To explore the effects of Ginkgo biloba extract EGB761 on the apoptosis related genes of neonatal rat cardiomyocyte induced by hypoxia and it’s molecular mechanism. Methods: Neonatal rat cardiomyocytes were isolated and cultured from 1 –dayold neonatal rats, and divided into control group, model group, EGB7615 μg/ml group, EGB761  10 μg/ml group and EGB761  20 μg/ml group. Cell viability was detected by MTT. The expression of LDH in cellular supernatant and the expression of SOD in cell homogenate were detected by elisa. The effect on apoptosis was detected by Hoechst33342. The mRNA and protein expression of PPARγ, NFκBp65, IκB, Bax, Bcl2 and Caspase3 were tested by PCR and western blot respectively. Results: The decreased cell viability was tested when cells induced by 6 h after hypoxia. Increased secretion of LDH, reduced expression of intracellular SOD and enhanced apoptosis induced by hypoxia were inversed significantly in the 10 μg/ml and 20 μg/ml EGB761 group (P<0.05). Decreased expression of PPARγ, IκB and increased expression of NFκB induced by hypoxia were inversed in different extent by the varying dose of EGB761 (P<0.05). In terms of apoptosis related gene, increased expression of Bax, Caspase3 and decreased expression of Bcl2 induced by hypoxia were also reversed by the varying dose of EGB761 (P<0.05). Conclusion:Ginkgo biloba extract EGB761 regulated the expression of PPARγ, NFκBp65, and apoptosis related genes, and reversed both the excessive myocardial cell apoptosis and the decreased survival rates caused by hypoxia.

参考文献/References:

［1］ Shen J, Lee W, Gu Y,et al. Ginkgo biloba extract (EGB761) inhibits mitochondriadependent caspase pathway and prevents apoptosis in hypoxiareoxygenated cardiomyocytes[J] . Chin Med, 2011,11(6):8.〖ZK)〗 ［2］ 闵宁斌, 唐治国, 黎 璞, 等. 银杏叶提取物对新生SD乳鼠心肌细胞缺氧损伤的影响[J] . 现代生物医学进展, 2012,12(18):34643468. ［3］ MahmoudAwny M, Attia AS, AbdEllah MF,et al. Mangiferin Mitigates Gastric Ulcer in Ischemia/ Reperfused Rats: Involvement of PPARγ, NFκB and Nrf2/HO1 Signaling Pathways[J] . PLoS One, 2015,10(7):e0132497. ［4］ 李 志. 银杏叶提取物金纳多(EGB761)[J] . 中国神经免疫学和神经病学杂志, 1994,1(2):121122. ［5］ 李淑琴, 朱嘉宝, 武宇洲. 银杏叶提取物防治心脑血管疾病的研究进展[J] . 中国新药杂志, 2016,25(1):7681. ［6］ Shi H, Zhang X, He Z,et al. Metabolites of Hypoxic Cardiomyocytes Induce the Migration of Cardiac Fibroblasts[J] . Cell Physiol Biochem, 2017,41(1):413421. ［7］ 郭长青, 郭 妍, 陈相健, 等. 银杏叶提取物EGB761对D半乳糖诱导的心肌细胞老化的保护作用[J] . 中国临床药理学与治疗学, 2010,15(4):376380. ［8］ Zhang Q, Shang M, Zhang M,et al. Microvesicles derived from hypoxia/reoxygenationtreated human umbilical vein endothelial cells promote apoptosis and oxidative stress in H9c2 cardiomyocytes[J] . BMC Cell Biol, 2016,17(1):25. ［9］ Li H, Song F, Duan LR,et al. Paeonol and danshensu combination attenuates apoptosis in myocardial infarcted rats by inhibiting oxidative stress: Roles of Nrf2/HO1 and PI3K/Akt pathway[J] . Sci Rep, 2016,6:23693.〖ZK)〗 ［10］ 赵艳荣, 张国斌, 邴飞虹. 银杏黄酮对心肌缺血再灌注损伤大鼠的心肌保护作用及其作用机制研究[J] . 实用心脑肺血管病杂志, 2016, 24(3):3842. ［11］ Domitrovi R, Cvijanovi O, Uni V,et al. Renoprotective mechanisms of chlorogenic acid in cisplatininduced kidney injury[J] . Toxicology, 2014,324:98107. ［12］ Yao J, Cao X, Zhang R,et al. Protective Effect of Baicalin Against Experimental Colitis via Suppression of Oxidant Stress and Apoptosis[J] . Pharmacogn Mag, 2016,12(47):225234. ［13］ Kiran KKM, Naveen KM, Patil RH,et al. Cadmium induces oxidative stress and apoptosis in lung epithelial cells[J] . Toxicol Mech Methods, 2016,26(9):658666. ［14］ Liu Y, Yi L, Wang L,et al. Ginsenoside Rg1 protects human umbilical cord bloodderived stromal cells against tertButyl hydroperoxideinduced apoptosis through AktFoxO3aBim signaling pathway[J] . Mol Cell Biochem, 2016,421(12):7587. ［15］ 陈春莲. NO,SOD,MDA与缺血性脑血管病的关系研究[J] . 广西医科大学学报, 2000,17(5):835836. ［16］ Zhang C, Wang DF, Zhang Z,et al. EGb 761 Protects Cardiac Microvascular Endothelial Cells against Hypoxia/Reoxygenation Injury and Exerts Inhibitory Effect on the ATM Pathway[J] . J Microbiol Biotechnol, 2017,27(3):584590. ［17］ Kukreja RC, Xi L. eNOS phosphorylation: a pivotal molecular switch in vasodilation and cardioprotection[J] . J Mol Cell Cardiol, 2007,42(2):280282. ［18］ Qiao ZY, Huang JH, Ma JW,et al. Ginkgo biloba extract reducing myocardium cells apoptosis by regulating apoptotic related proteins expression in myocardium tissues[J] . Mol Biol Rep, 2014,41(1):347353. ［19］ Lee NJ, Oh JH, Ban JO,et al. 4Omethylhonokiol, a PPARγ agonist, inhibits prostate tumour growth: p21mediated suppression of NFκB activity[J] . Br J Pharmacol, 2013,168(5):11331145. ［20］ Lützen U, Zhao Y, Lucht K,et al. Pioglitazone induces cell growth arrest and activates mitochondrial apoptosis in human uterine leiomyosarcoma cells by a peroxisome proliferatoractivated receptor γindependent mechanism[J] . Naunyn Schmiedebergs Arch Pharmacol, 2017,390(1):3748. ［21］ Zeng XC, Li L, Wen H,et al. MicroRNA128 inhibition attenuates myocardial ischemia/reperfusion injuryinduced cardiomyocyte apoptosis by the targeted activation of peroxisome proliferatoractivated receptor gamma[J] . Mol Med Rep, 2016,14(1):129136. ［22］ Suzuki T, Saiki Y, Horii A,et al. Atrial natriuretic peptide induces peroxisome proliferator activated receptor γ during cardiac ischemiareperfusion in swine heart[J] . Gen Thorac Cardiovasc Surg, 2017,65(2):8595. ［23］ Han J, Wang D, Ye L,et al. Rosmarinic Acid Protects against Inflammation and Cardiomyocyte Apoptosis during Myocardial Ischemia/Reperfusion Injury by Activating Peroxisome ProliferatorActivated Receptor Gamma[J] . Front Pharmacol, 2017,25(8):456. ［24］ Neri T, Armani C, Pegoli A,et al. Role of NFkappaB and PPARgamma in lung inflammation induced by monocytederived microparticles[J] . Eur Respir J, 2011,37(6):14941502. ［25］ 杨靖宇. 沉默PPARγ通过上调bcl2表达抑制A549细胞凋亡[J] . 中国肺癌杂志, 2013,16(3):125130.

更新日期/Last Update: 2019-03-13