CAJ | 학술논문

目的探讨亲环素A(CyPA)对Aβ25-35诱导的PC12细胞凋亡影响及可能的作用机制.方法将PC12细胞分为正常对照组(0 μmol/L Aβ25-35)、Aβ25-35诱导组(10 μmol/L Aβ25-35)和药物保护组(0.1、1、10、100 nmol/L CyPA+10 μmoi/Lβ25-35),药物保护组采用相应浓度CyPA预处理PC12细胞30 min,再加入Aβ25-35继续培养.MTT法分析细胞存活率,Hoechst33258染色法检测细胞凋亡,RT-PCR检测Bcl-2和Bax mRNA表达,Western blotting检测Bcl-2和Bax蛋白表达.结果 1、10和100 nmol/L CyPA可提高细胞的存活率,减少Aβ25-35引起的细胞凋亡,增加Bcl-2mRNA表达以及Bcl-2蛋白表达,降低Bax mRNA表达以及Bax蛋白表达,与Aβ25-35诱导组比较差异有统计学意义(P<0.05),且CyPA剂量越高效果越明显.结论 CyPA可剂量依赖性对抗Aβ25-35对PC12细胞的毒性作用,减少细胞凋亡,其机制与上调抗凋亡基因Bcl-2表达和下调凋亡基因Bax表达有关.
목적 탐토친배소A(CyPA)대Aβ25-35유도적PC12세포조망영향급가능적작용궤제.방법 장PC12세포분위정상대조조(0 μmol/L Aβ25-35)、Aβ25-35유도조(10 μmol/L Aβ25-35)화약물보호조(0.1、1、10、100 nmol/L CyPA+10 μmoi/Lβ25-35),약물보호조채용상응농도CyPA예처리PC12세포30 min,재가입Aβ25-35계속배양.MTT법분석세포존활솔,Hoechst33258염색법검측세포조망,RT-PCR검측Bcl-2화Bax mRNA표체,Western blotting검측Bcl-2화Bax단백표체.결과 1、10화100 nmol/L CyPA가제고세포적존활솔,감소Aβ25-35인기적세포조망,증가Bcl-2mRNA표체이급Bcl-2단백표체,강저Bax mRNA표체이급Bax단백표체,여Aβ25-35유도조비교차이유통계학의의(P<0.05),차CyPA제량월고효과월명현.결론 CyPA가제량의뢰성대항Aβ25-35대PC12세포적독성작용,감소세포조망,기궤제여상조항조망기인Bcl-2표체화하조조망기인Bax표체유관.
Objective To explore the effect of cyclophilin A (CyPA) on apoptosis of PC 12 cells induced by Aβ25-35 and its potential mechanism. Methods PC 12 cells were divided into normal control group (0 μmol/L Aβ25-35), Aβ25-35 inducement group (10 μmol/L Aβ25-35) and drug protection groups (0.1, 1,10 and 100 nmol/L CyPA+10 μmol/L Aβ25-35). Cells in the drug protection groups were pretreated by CyPA of different concentrations for 30 min, and then co-cultured with Aβ25-35 We evaluated the survival rate of PC12 cells with MTT assay, analyzed the apoptosis of PC12 cells with Hoechst33258 staining, and detected the mRNA expressions of Bcl-2 and Bax with PT-PCR and the protein levels of Bcl-2 and Bax with Western blotting. Results Cells pretreated wth CyPA of 1, 10 and 100 nmol/L enjoyed an obvious elevation of survival rate of PC 12 cells, a significant reduction of apoptosis induced by Aβ25-35,an obvious increase of mRNA expression of Bcl-2 and protein level of Bcl-2, and a statistical decrease of mRNA expression of Bax and protein level of Bax as compared with those cells of the Aβ25-35 inducement group (P<0.05);and these effects were dose-dependent. Conclusion CyPA could resist the toxic role of Aβ25-35 on PC 12 cells and reduce the apoptosis in a dose-dependent manner by up-regulation of anti-apoptosis gene Bcl-2 and down-regulation of apoptosis gene Bax.