CAJ | 학술논문

背景:目前对抗氧化基因硫氧还蛋白的研究逐步受到重视,但从基因治疗的角度对其研究较少.目的:采用硫氧还蛋白转染 Neuro-2A 细胞,观察细胞内表达相应蛋白因子对细胞的具体保护效果,并分析其发挥保护作用的可能机制.方法:以质粒 PIRES2-EGFP-TRX 转染 Neuro-2A 细胞,经 RT-PCR 鉴定,建立能够稳定表达硫氧还蛋白的细胞株.利用不同浓度的 H2O2处理正常细胞及转染细胞,建立氧化应激模型.观察受到氧化损伤后两组细胞的形态学、存活率、还原型谷胱甘肽的浓度及细胞内 DNA 链断裂情况.结果与结论:经 H2O2作用后,两组细胞形态均出现损伤,但转染组细胞比正常组细胞损伤减轻、细胞存活率升高、细胞内还原型谷胱甘肽水平增高、DNA 链断裂程度减轻.说明人类硫氧还蛋白基因可以在 Neuro-2A 细胞中被重组并顺利表达,对细胞具有一定的保护作用;这一作用可能是通过清除氧自由基,维持细胞内还原型谷胱甘肽水平,从而保护细胞 DNA 免受氧化性损伤来实现的.
배경:목전대항양화기인류양환단백적연구축보수도중시,단종기인치료적각도대기연구교소.목적:채용류양환단백전염 Neuro-2A 세포,관찰세포내표체상응단백인자대세포적구체보호효과,병분석기발휘보호작용적가능궤제.방법:이질립 PIRES2-EGFP-TRX 전염 Neuro-2A 세포,경 RT-PCR 감정,건립능구은정표체류양환단백적세포주.이용불동농도적 H2O2처리정상세포급전염세포,건립양화응격모형.관찰수도양화손상후량조세포적형태학、존활솔、환원형곡광감태적농도급세포내 DNA 련단렬정황.결과여결론:경 H2O2작용후,량조세포형태균출현손상,단전염조세포비정상조세포손상감경、세포존활솔승고、세포내환원형곡광감태수평증고、DNA 련단렬정도감경.설명인류류양환단백기인가이재 Neuro-2A 세포중피중조병순리표체,대세포구유일정적보호작용;저일작용가능시통과청제양자유기,유지세포내환원형곡광감태수평,종이보호세포 DNA 면수양화성손상래실현적.
BACKGROUND: Studies on antioxidant genes Thioredoxin (TRX) have attracted more attention gradual y, but relevant studies from the aspect of gene therapy are fewer. OBJECTIVE: To study the protective effect of cells expressing the corresponding proteins in TRX transfected Neuro-2A cells and to analyze the possible mechanism underlying the protective effect. METHODS: Plasmids PIRES2-EGFP-TRX were used to transfect Neuro-2A cells and to establish a cel line which could stably express TRX proteins identified by reverse transcription-PCR. Different concentrations of hydrogen peroxide were used to treat normal cells and transfected cells for the establishment of oxidative stress models. Cel morphology, cellsurvival, glutathione concentration and DNA strand breaks situation were observed after oxidative damage. RESULTS AND CONCLUSION: Normal cells and transfected cells were both damaged by hydrogen peroxide; however, the transfected cells were superior to the normal cells in cel damage, cellsurvival, intracel ular glutathione level and degree of DNA strand breaks. These findings indicate that TRX gene in Neuro-2A cells can be reconstructed and expressed successful y, which plays a certain protective. This effect may be achieved through scavenging oxygen free radicals, maintaining intracel ular glutathione levels, thereby protecting cel ular DNA against oxidative damage.