首页 | 本学科首页   官方微博 | 高级检索  
     

过表达ODC抑制氧化应激诱导大鼠心肌细胞凋亡的研究
引用本文:臧书文,刘彬,周迎春,孙学刚,黄平,邱锦帆,刘紫庭. 过表达ODC抑制氧化应激诱导大鼠心肌细胞凋亡的研究[J]. 热带医学杂志, 2012, 12(5): 510-513,522,639
作者姓名:臧书文  刘彬  周迎春  孙学刚  黄平  邱锦帆  刘紫庭
作者单位:1. 南方医科大学中医药学院,广东广州,510515
2. 南方医科大学中医药学院,广东广州510515;南方医科大学中西医结合医院心内科,广东广州510450;南方医科大学南方医院中医科,广东广州510515
3. 南方医科大学中西医结合医院心内科,广东广州,510450
4. 南方医科大学南方医院中医科,广东广州,510515
基金项目:国家自然科学基金(30973850、81173459);广东省科技计划项目(2010B060500009)
摘    要:目的构建过表达鸟氨酸脱羧酶(ODC)基因慢病毒载体,并研究其对氧化应激诱导大鼠H9C2心肌细胞凋亡的影响。方法构建大鼠ODC基因pHIV-ODC过表达质粒,与包装质粒psPAX2、pMD2G共转染293FT细胞,检测其侵染效率;包装慢病毒并侵染H9C2心肌细胞;72 h后观察其侵染效率,RT-PCR法检测细胞ODC mRNA的表达,利用CCK-8、Hochest33258染色检测ODC对H2O2诱导H9C2心肌细胞凋亡的影响。结果酶切及测序鉴定ODC序列正确插入慢病毒过表达载体。侵染H9C2细胞72 h后ODC mRNA水平较阴性慢病毒感染组显著升高。过表达ODC能抑制H2O2诱导的细胞活力下降,减轻H2O2诱导的心肌细胞凋亡,与H2O2组比较差异均有统计学意义(P<0.001)。结论成功构建的过表达ODC慢病毒,上调H9C2细胞中ODC的表达,过表达ODC能显著抑制氧化应激诱导的H9C2心肌细胞凋亡。

关 键 词:鸟氨酸脱羧酶  心肌细胞凋亡  氧化应激

Overexpression of ornithine decarboxylase attenuates oxidative stress-induced apoptosis in H9C2 cells
ZANG Shu-wen , LIU Bin , ZHOU Ying-chun , SUN Xue-gang , HUANG Ping , QIU Jin-fan , LIU Zi-ting. Overexpression of ornithine decarboxylase attenuates oxidative stress-induced apoptosis in H9C2 cells[J]. Journal Of Tropical Medicine, 2012, 12(5): 510-513,522,639
Authors:ZANG Shu-wen    LIU Bin    ZHOU Ying-chun    SUN Xue-gang    HUANG Ping    QIU Jin-fan    LIU Zi-ting
Affiliation:1(1.School of Traditional Chinese Medicine,Southern Medical University,Guangdong,Guangzhou 510515; 2.Cardiology Department of Integrated Traditional Chinese Medical and Western Medicine Hospital,Southern Medical University,Guangdong,Guangzhou 510450;3.Department of Traditional Chinese Medicine,Nanfang Hospital,Southern Medical University,Guangdong,Guangzhou 510515,China)
Abstract:Objective To construct the lentiviral overexpression vector of ornithine decarboxylase(ODC) and investigate its role in oxidative stress-induced apoptosis in H9C2 cells.Methods The lentiviral vector(pHIV) was constructed by inserting the lentiviral vectors with the ODC gene fragment.The recombination lentiviral particles were produced by the packaging 293T cell,the culture supernatant was harvested and the titration of them was determined by the limiting dilution.H9C2 cells were infected with recombinant lentivirus overexpressing ODC,after 72 hours.The expression level of ODC in cells was examined by the RT-PCR.The effects of ODC overexpression on oxidative stress-induced apoptosis in H9C2 cell were investigated by CCK-8 assay and Hoechst33258 staining.Results The transduction efficiency of H9C2 cell line with pHIV-ODC lentivirus was 95%.72 h after virus infection,ODC mRNA levels were higher than those of negative control group.Compare with H2O2-induced apoptotic cells,cell viability and the amount of apoptotic cells in the ODC overexpressed H9C2 cells was significantly decreased(P<0.001).Conclusion ODC can be overexpressed in H9C2 cells by recombinant lentivirus infection,which leads to a significant inhibition of oxidative stress-induced apoptosis in these cells.
Keywords:ornithine decarboxylase  apoptosis  oxidative stress
本文献已被 CNKI 万方数据 等数据库收录!
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号