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Heat shock protein 70 gene transfection protects rat myocardium cell against anoxia-reoxygeneration injury

Chinese Medical Journal 2007年第7期120卷 578-583页

作者： LIU Ji-chun HE Ming WAN Li CHENG Xiao-shu Department of Cardiovascular Surgery First Affiliated Hospital of Nanchang University Nanchang 330006 China Department of Pharmacology Medical College of Nanchang University Nanchang 330006 China Department of Cardiovascular Medicine Second Affiliated Hospital of Nanchang University Nanchang 330006 China

Background A number of studies suggest that the expression of heat shock protein 70 （hsp70） induced by heat stress are associated with protection against ischemia-reperfusion injury. But the protective effects may be contaminated by other factors in the same stress. This study was conducted to explore the protective role of hsp70 expression in acute myocardial anoxia/reoxygeneration （A/R） injury with a liposome-mediated gene transfer technique for the introduction of pCDNA hsp70 into the neonatal rat myocardial cells. In addition, heat shock stress cytoprotection was also investigated for comparison. Methods The cultured primary neonatal rat myocardiocytes with an acute myocardial A/R injury model and the HS-treated rat myocardiocyte model were used. Three-day cultured myocardiocytes were randomly divided into four groups （n=8）： control group, A/R group, HS＋A/R group and pCDNA hsp70 ＋A/R group. A liposome-coated hsp70 pCDNA plasmid was transfected into the primary neonatal rat myocardiocytes; hsp70 mRNA and its protein were confirmed by reverse transcriptase polymerase chain reaction （RT-PCR） and Western blotting. The cell viability was assayed by monotetrazolium （MTT） and the lactate dehydrogenase （LDH） and creatine phosphokinase （CPK） activity of cells during incubation and the changes in cells ultrastructure were examined. NF-κB activity in the primary neonatal rat myocardiocytes was measured with flow cytometry. Results Compared with viability in the A/R group （（35.4±6.9）%） the cell viability in the HS＋A/R group （（72.8±11.6）%） and the pCDNA hsp70 ＋ A/R group （（76.3±12.2）%） was improved significantly （P〈0.05）. The activity of LDH and CPK was significantly elevated in the A/R group. However, in the HS＋A/R group and pCDNA hsp70 ＋A/R group, significant decreases in activity were observed. The cell ultrastructure of the A/R group cells was abnormal, whereas nearly normal ultrastructure was observed in HS＋A/R group and pCDNA hsp70＋A/R group. hsp70 mRNA and protein

关键词： gene transfection hsp70 gene NF-κB cardiac myocyte anoxia-reoxygeneration injury

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