中国全科医学
中國全科醫學
중국전과의학
CHINESE GENERAL PRACTICE
2010年
5期
494-497
,共4页
宋晓聪%杨芳芳%胡丹%曲彦
宋曉聰%楊芳芳%鬍丹%麯彥
송효총%양방방%호단%곡언
热休克蛋白70%腺病毒载体%过氧化氢%神经元
熱休剋蛋白70%腺病毒載體%過氧化氫%神經元
열휴극단백70%선병독재체%과양화경%신경원
Heat shock proteins 70%Adenovirus%Hydrogen peroxide%Neuronal cells
目的 探讨重组腺病毒介导的HSP70表达对过氧化氢(H_2O_2)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、Western 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H_2O_2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.H_2O_2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H_2O_2损伤,具有明确的细胞保护作用.
目的 探討重組腺病毒介導的HSP70錶達對過氧化氫(H_2O_2)誘導的神經元和膠質細胞損傷的保護作用.方法 用攜帶全長HSP70基因的重組腺病毒vAd-HSP70感染體外培養的神經元和膠質細胞,RT-PCR、Western blotting檢測靶細胞中外源性HSP70的錶達.vAd-HSP70感染組、vAd-GFP感染組和未感染組細胞經0.5 mmol/L的H_2O_2處理後,MTT檢測細胞活力,乳痠脫氫酶(LDH)檢測試劑盒檢測細胞培養上清液中LDH活力,透射電鏡觀察細胞超微結構變化.結果 vAd-HSP70感染組的細胞可檢測到外源性HSP70基因錶達.H_2O_2處理後,vAd-HSP70感染組細胞活力較其他組明顯增彊(P<0.01),vAd-HSP70感染組、vAd-GFP感染組、未感染組細胞培養上清液中LDH活力分彆為(976±106)、(1 332±197)、(1 380±121),差異有統計學意義(P<0.01).電鏡結果顯示,vAd-HSP70感染組細胞膜較vAd-GFP感染組和未感染組完整清晰,無明顯線粒體腫脹及細胞覈溶解等不可逆損傷情況.結論 腺病毒介導的外源性HSP70錶達可保護神經元和膠質細胞牴抗H_2O_2損傷,具有明確的細胞保護作用.
목적 탐토중조선병독개도적HSP70표체대과양화경(H_2O_2)유도적신경원화효질세포손상적보호작용.방법 용휴대전장HSP70기인적중조선병독vAd-HSP70감염체외배양적신경원화효질세포,RT-PCR、Western blotting검측파세포중외원성HSP70적표체.vAd-HSP70감염조、vAd-GFP감염조화미감염조세포경0.5 mmol/L적H_2O_2처리후,MTT검측세포활력,유산탈경매(LDH)검측시제합검측세포배양상청액중LDH활력,투사전경관찰세포초미결구변화.결과 vAd-HSP70감염조적세포가검측도외원성HSP70기인표체.H_2O_2처리후,vAd-HSP70감염조세포활력교기타조명현증강(P<0.01),vAd-HSP70감염조、vAd-GFP감염조、미감염조세포배양상청액중LDH활력분별위(976±106)、(1 332±197)、(1 380±121),차이유통계학의의(P<0.01).전경결과현시,vAd-HSP70감염조세포막교vAd-GFP감염조화미감염조완정청석,무명현선립체종창급세포핵용해등불가역손상정황.결론 선병독개도적외원성HSP70표체가보호신경원화효질세포저항H_2O_2손상,구유명학적세포보호작용.
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 (H_2O_2),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 H_2O_2 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 H_2O_2 injuries,with obvious cell protective effects.
