CAJ | 학술논문

目的探讨3-硝基丙酸(3-NP)多次化学预处理对多巴胺能神经元的保护作用及可能机制.方法应用MPTP(30 mg/kg)在C57BL小鼠上复制帕金森病模型,以3-NP(20 mg/kg)行预处理,检测小鼠中脑黑质凋亡率和转录因子c-Jun的阳性细胞数量及c-Jun的蛋白水平;应用MPP+(0.25 mmol/L)在SH-SY5Y细胞制作帕金森病模型,以3-NP(0.2 mmol/L)进行预处理,并将携带显性突变体c-JuncDNA片段的真核表达载体质粒pcDNA3(HA)-Jun-dn转染SH-SY5Y细胞,检测各组细胞的c-Jun表达水平及凋亡率.结果小鼠中脑黑质凋亡率:MPTP组较对照组明显升高(P＜0.01),3-NP单次、多次预处理后均明显降低(P＜0.05,P＜0.01);c-Jun阳性细胞数:MPTP组较对照组明显增加(P＜0.05),3-NP单次预处理组与MPTP组比较无明显差异,3-NP多次预处理后明显降低(P＜0.05);c-Jun蛋白水平:与其阳性细胞数变化一致;细胞凋亡率:MPP+组较对照组明显升高,3-NP单次、多次预处理组细胞凋亡率明显降低(P＜0.05,P＜0.01);c-Jun蛋白水平变化与中脑黑质一致;经pcDNA3(HA)-Jun-dn转染的细胞,其c-Jun的表达较未转染细胞明显降低(P＜0.01),其凋亡率也下降(P＜0.01).结论3-NP单次、多次预处理对多巴胺能神经元确有保护作用,多次预处理保护效果更强,其机制与抑制转录因子c-Jun的表达,降低其蛋白水平有关.
목적탐토3-초기병산(3-NP)다차화학예처리대다파알능신경원적보호작용급가능궤제.방법응용MPTP(30 mg/kg)재C57BL소서상복제파금삼병모형,이3-NP(20 mg/kg)행예처리,검측소서중뇌흑질조망솔화전록인자c-Jun적양성세포수량급c-Jun적단백수평;응용MPP+(0.25 mmol/L)재SH-SY5Y세포제작파금삼병모형,이3-NP(0.2 mmol/L)진행예처리,병장휴대현성돌변체c-JuncDNA편단적진핵표체재체질립pcDNA3(HA)-Jun-dn전염SH-SY5Y세포,검측각조세포적c-Jun표체수평급조망솔.결과소서중뇌흑질조망솔:MPTP조교대조조명현승고(P＜0.01),3-NP단차、다차예처리후균명현강저(P＜0.05,P＜0.01);c-Jun양성세포수:MPTP조교대조조명현증가(P＜0.05),3-NP단차예처리조여MPTP조비교무명현차이,3-NP다차예처리후명현강저(P＜0.05);c-Jun단백수평:여기양성세포수변화일치;세포조망솔:MPP+조교대조조명현승고,3-NP단차、다차예처리조세포조망솔명현강저(P＜0.05,P＜0.01);c-Jun단백수평변화여중뇌흑질일치;경pcDNA3(HA)-Jun-dn전염적세포,기c-Jun적표체교미전염세포명현강저(P＜0.01),기조망솔야하강(P＜0.01).결론3-NP단차、다차예처리대다파알능신경원학유보호작용,다차예처리보호효과경강,기궤제여억제전록인자c-Jun적표체,강저기단백수평유관.
Objective To investigate the protective effect of 3-NP repetitive preconditioning on dopaminergic neurons.Methods Parkinson' s disease model were made on C57BL mouse with MPTP ( 30 mg/kg). 3-NP were administered to produce preconditioning before MPTP used. The apoptosis ratio and expression of c-Jun in the substantia nigra were assayed. MPP + were used to made parkinson' s disease model in vitro. 3-NP were incubated to produce preconditioning;The fragment of dominant mutant c-JuncDNA carried by eukaryon plasmid pcDNA3 (HA)-jun-dn was transfected into SHSY5Y cells. The expression of c-Jun and the apoptosis ratio of SH-SY5Y cells were assayed. Results In MPTP group,the ratio of apoptosis was increased significantly. After 3-NP preconditioning, apoptosis was decreased significantly and the effect of repetitive preconditioning was better than that of single preconditioning. The expression of c-Jun was increased in MPTP group and no obviously changes were observed after 3-NP single preconditioning, but decreased after 3-NP repetitive preconditioning. In the transfection group, the expression of c-Jun was decreased and the apoptosis ratio was lower than that without transfection group. 3-NP repetitive preconditioning could decrease the expression of c-Jun and apoptosis ratio. Conclusion 3-NP preconditioning exerts protective effect on dopaminergic neurons and the effect of repetitive preconditioning was better than single preconditioning.