摘要
目的探讨重组腺病毒介导的HSP70表达对过氧化氢(H2O2)诱导的神经元和胶质细胞损伤的保护作用。方法用携带全长HSP70基因的重组腺病毒vAd-HSP70感染体外培养的神经元和胶质细胞,RT-PCR、West-ern blotting检测靶细胞中外源性HSP70的表达。vAd-HSP70感染组、vAd-GFP感染组和未感染组细胞经0.5 mmol/L的H2O2处理后,MTT检测细胞活力,乳酸脱氢酶(LDH)检测试剂盒检测细胞培养上清液中LDH活力,透射电镜观察细胞超微结构变化。结果vAd-HSP70感染组的细胞可检测到外源性HSP70基因表达。H2O2处理后,vAd-HSP70感染组细胞活力较其他组明显增强(P<0.01),vAd-HSP70感染组、vAd-GFP感染组、未感染组细胞培养上清液中LDH活力分别为(976±106)、(1 332±197)、(1 380±121),差异有统计学意义(P<0.01)。电镜结果显示,vAd-HSP70感染组细胞膜较vAd-GFP感染组和未感染组完整清晰,无明显线粒体肿胀及细胞核溶解等不可逆损伤情况。结论腺病毒介导的外源性HSP70表达可保护神经元和胶质细胞抵抗H2O2损伤,具有明确的细胞保护作用。
Objective To investigate the protective effects of recombinant adenovirus-mediated heat shock protein 70（HSP70） on neurons and glial cell injury induced by oxidative stress.Methods Cultured neurons and glial cells were divided into groups A（vAd-HSP70 infected,carrying whole long human HSP70 gene）,B（vAd-GFP infected） and C（non-infected）.After cells were treated by 0.5 mmol / L hydrogen peroxide（H2O2）,cell viability was detected by MTT,lactate dehydrogenase（LDH） activity in cell culture supernatant by LDH detection kit,the changes of cell ultrastructure observed with transmission electron microscopy.Results The expression of ectogenous HSP 70 gene was detected by RT-PCR and Western blotting in group A.Cell viability of group A was significantly higher than that of groups B,C after H2O2 treatment（P〈0.01）.LDH activities in cell culture supernatant of groups A,B,C were 976±106,1 332±197,1 380±121,respectively,the difference was significant（P〈0.01）.The results of transmission electron microscopy showed that the cytomembrane of group A was more complete and clearer than the other 2 groups,without any irreversible injuries such as obvious mitochondrial swelling or nuclear dissolution.Conclusion The expression of adenovirus-induced exogenous HSP 70 gene can prevent neurons and glial cells from H2O2 injuries,with obvious cell protective effects.
出处
《中国全科医学》
CAS
CSCD
北大核心
2010年第5期494-497,共4页
Chinese General Practice
基金
山东省自然科学基金资助项目(Y2007C133)
青岛市市南区科学技术局资助项目
