CAJ | 학술논문

目的研究朊蛋白106-126肽段在细胞水平上的毒性作用机制.方法 PC12细胞常规培养后加入神经生长因子(NGF)诱导成神经元样细胞,再加入朊蛋白106-126肽段,检测细胞存活率,观察细胞生长和形态变化,检测细胞氧化应激现象和能量代谢的改变,检测细胞凋亡现象及其相关机制.结果细胞接触肽段后存活率由(98.1±1.9)%下降到(69.2±4.7)%,流式细胞仪检测细胞出现亚二倍体峰.氧化应激现象于接触肽段12 h后出现并且持续存在,细胞内钙离子浓度从(185.74±12.93)nmol:L增加到(493.00±58.71)nmol/L,线粒体膜电位下降至65%,Ca2+ATP酶活性从54.92±4.05降低至34.92±4.86,细胞能量代谢水平下降,最终出现细胞凋亡,Bcl-2/Bax系统稳态变化参与了诱导凋亡的过程.结论利用106-126肽段感染分化的PC12细胞可能是在细胞水平上研究朊蛋白毒性作用的理想模型.PrP106-126的毒性作用是通过持续的氧化应激、干扰细胞的能量代谢、钙超载、最终诱导宿主细胞的凋亡来实现的,并且氧化应激可能处于始动与核心的地位.
목적 연구원단백106-126태단재세포수평상적독성작용궤제.방법 PC12세포상규배양후가입신경생장인자(NGF)유도성신경원양세포,재가입원단백106-126태단,검측세포존활솔,관찰세포생장화형태변화,검측세포양화응격현상화능량대사적개변,검측세포조망현상급기상관궤제.결과 세포접촉태단후존활솔유(98.1±1.9)%하강도(69.2±4.7)%,류식세포의검측세포출현아이배체봉.양화응격현상우접촉태단12 h후출현병차지속존재,세포내개리자농도종(185.74±12.93)nmol:L증가도(493.00±58.71)nmol/L,선립체막전위하강지65%,Ca2+ATP매활성종54.92±4.05강저지34.92±4.86,세포능량대사수평하강,최종출현세포조망,Bcl-2/Bax계통은태변화삼여료유도조망적과정.결론 이용106-126태단감염분화적PC12세포가능시재세포수평상연구원단백독성작용적이상모형.PrP106-126적독성작용시통과지속적양화응격、간우세포적능량대사、개초재、최종유도숙주세포적조망래실현적,병차양화응격가능처우시동여핵심적지위.
Objective To investigate the cytotoxic effects of differentiated PC12 cells afterinfected by prion protein 106-126 peptide.Methods The PC12 cells were infected by prion protein 106-126peptide after differentiated by nerve growthfactor(NGF).Cell viability andthe morphological changes were observed.The energy metabolize and apoptosis was detected.Results Afterinfected by this peptide,cell viability decreasedfrom(98.1±1.9)% to (69.2±4.7)%,and apoptosis peak Was observed byflow cytometry.Aboutthe process of the cytotoxic effects,afterthe cells affected by PrP106-126,oxidative stress presented and existed continually,and then the intracellular free calcium concentrate increased from (185.74±12.93)nmol/L to (493.00±58.71)nmol/L subsequently,the activity of Ca2+ ATPase decreased from 54.92±4.05 to 34.92±4.86,the mitochondrial membrane potential decreasedto 65%,and also the energy metabolize disorder,the cells presented apoptosisinthe end.The changed Bcl-2/Bax system involvedinthe apoptosis.Conclusions Prion protein106-126 peptide caninduce apoptosisin differentiated PC12 cells and presented cellulartoxicity definitely.It might be a perfect model to study the cellular toxicity of prion protein.Continual oxidative stress could causetheintracellularfree calcium concentrate and disturb the energy metabolize,and the apoptosis might be the end-result.The oxidative stress of might play a startup and important role.