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1d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Westernblot法检测PPAR-γ、NF-κB-p65、I-κB、Bax、Bcl-2、Caspase-3mRNA及蛋白表达。结果:和对照组相比,缺氧6h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。
Objective:To explore the effects of Ginkgo biloba extract EGB761 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 from1-day-old neonatal rats,and divided into control group,model group,EGB761-5μg/ml group,EGB761-10μg/ml group and EGB761-20μg/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-2and Caspase-3were tested by PCR and western blot respectively.Results:The decreased cell viability was tested when cells induced by6h after hypoxia.Increased secretion of LDH,reduced expression of intracellular SOD and enhanced apoptosis induced by hypoxia were inversed significantly in the10μg/ml and20μg/ml EGB761group(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 EGB761(P<0.05).In terms of apoptosis related gene,increased expression of Bax,Caspase-3and decreased expression of Bcl-2induced by hypoxia were also reversed by the varying dose of EGB761(P<0.05).Conclusion:Ginkgo biloba extract EGB761regulated 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.
作者
李鹏盈
卢成志
周小波
王桂贤
LI Pengying;LU Chengzhi;ZHOU Xiaobo(Department of Emergency, Dongli Hospital of Dongli District in Tianjin ,Tianjin 300300)
出处
《陕西中医》
2019年第1期3-8,共6页
Shaanxi Journal of Traditional Chinese Medicine
