中华神经科杂志
中華神經科雜誌
중화신경과잡지
Chinese Journal of Neurology
2013年
8期
546-550
,共5页
鲍兵%陈志颖%柴竞艳%周君%吴丹%殷小平
鮑兵%陳誌穎%柴競豔%週君%吳丹%慇小平
포병%진지영%시경염%주군%오단%은소평
硫氰酸盐类%PC12细胞%NF-E2相关因子2%1-甲基-4-苯基吡啶%氧化性应激
硫氰痠鹽類%PC12細胞%NF-E2相關因子2%1-甲基-4-苯基吡啶%氧化性應激
류청산염류%PC12세포%NF-E2상관인자2%1-갑기-4-분기필정%양화성응격
Thiocyanates%PC12 cells%NF-E2-related factor 2%1-Methyl-4-phenylpyridinium%Oxidative stress
目的 探讨莱菔硫烷对体外1-甲基-4-苯基吡啶离子(MPP+)诱导的PC12细胞氧化损伤模型是否具有保护作用及其机制.方法 以MPP+损伤PC12细胞制备氧化损伤模型.不同浓度的莱菔硫烷加入培养基观察各组细胞生长情况.后续实验分成4组:正常对照组(A)、莱菔硫烷组(B)、MPP+损伤组(C)、莱菔硫烷预处理+MPP+损伤组(D).通过MTT比色法检测细胞活力,观察不同浓度莱菔硫烷预处理PC12细胞活力变化及不同分组PC12细胞活力变化；流式细胞术检测不同分组PC12细胞中细胞凋亡率的变化；免疫印迹法测定不同分组PC12细胞内转录因子相关因子2(Nrf2)、血红素加氧酶1(HO-1)及醌还原氧化酶1(NQ01)蛋白的表达变化.结果 A组细胞存活率(98.70％)与MPP+(500μmol/L)组(58.16％)相比差异有统计学意义(F =21.83,P＜0.05),D组细胞存活率明显提高.C组细胞凋亡率升高,D组细胞与C组相比差异有统计学意义,莱菔硫烷预处理后细胞凋亡率明显减低.C组Nrf2、HO-1、NQ01蛋白表达下降,D组Nrf2、HO-1、NQ01蛋白表达升高.结论 莱菔硫烷对MPP+诱导的PC12细胞损伤具有保护作用,莱菔硫烷对MPP+诱导的PC12细胞损伤的保护作用可能通过激活Nrf2-抗氧化反应元件通路途径实现.
目的 探討萊菔硫烷對體外1-甲基-4-苯基吡啶離子(MPP+)誘導的PC12細胞氧化損傷模型是否具有保護作用及其機製.方法 以MPP+損傷PC12細胞製備氧化損傷模型.不同濃度的萊菔硫烷加入培養基觀察各組細胞生長情況.後續實驗分成4組:正常對照組(A)、萊菔硫烷組(B)、MPP+損傷組(C)、萊菔硫烷預處理+MPP+損傷組(D).通過MTT比色法檢測細胞活力,觀察不同濃度萊菔硫烷預處理PC12細胞活力變化及不同分組PC12細胞活力變化；流式細胞術檢測不同分組PC12細胞中細胞凋亡率的變化；免疫印跡法測定不同分組PC12細胞內轉錄因子相關因子2(Nrf2)、血紅素加氧酶1(HO-1)及醌還原氧化酶1(NQ01)蛋白的錶達變化.結果 A組細胞存活率(98.70％)與MPP+(500μmol/L)組(58.16％)相比差異有統計學意義(F =21.83,P＜0.05),D組細胞存活率明顯提高.C組細胞凋亡率升高,D組細胞與C組相比差異有統計學意義,萊菔硫烷預處理後細胞凋亡率明顯減低.C組Nrf2、HO-1、NQ01蛋白錶達下降,D組Nrf2、HO-1、NQ01蛋白錶達升高.結論 萊菔硫烷對MPP+誘導的PC12細胞損傷具有保護作用,萊菔硫烷對MPP+誘導的PC12細胞損傷的保護作用可能通過激活Nrf2-抗氧化反應元件通路途徑實現.
목적 탐토래복류완대체외1-갑기-4-분기필정리자(MPP+)유도적PC12세포양화손상모형시부구유보호작용급기궤제.방법 이MPP+손상PC12세포제비양화손상모형.불동농도적래복류완가입배양기관찰각조세포생장정황.후속실험분성4조:정상대조조(A)、래복류완조(B)、MPP+손상조(C)、래복류완예처리+MPP+손상조(D).통과MTT비색법검측세포활력,관찰불동농도래복류완예처리PC12세포활력변화급불동분조PC12세포활력변화；류식세포술검측불동분조PC12세포중세포조망솔적변화；면역인적법측정불동분조PC12세포내전록인자상관인자2(Nrf2)、혈홍소가양매1(HO-1)급곤환원양화매1(NQ01)단백적표체변화.결과 A조세포존활솔(98.70％)여MPP+(500μmol/L)조(58.16％)상비차이유통계학의의(F =21.83,P＜0.05),D조세포존활솔명현제고.C조세포조망솔승고,D조세포여C조상비차이유통계학의의,래복류완예처리후세포조망솔명현감저.C조Nrf2、HO-1、NQ01단백표체하강,D조Nrf2、HO-1、NQ01단백표체승고.결론 래복류완대MPP+유도적PC12세포손상구유보호작용,래복류완대MPP+유도적PC12세포손상적보호작용가능통과격활Nrf2-항양화반응원건통로도경실현.
Objective To investigate the effect of sulforaphane on 1-methyl-4-phenylpyridinium (MPP +)-induced cell viability loss in cultured PC12 cells and to explore the possible mechanism.Methods MPP + induced damage in PC12 cells was prepared as oxidative damage model.Sulforaphane (0.5,1.O,2.5,5.0 and 10.0 μmol/L) was added in each group cell growth medium.Subsequent experiments were divided into 4 groups:(A) normal control group,(B) sulforaphane group,(C) MPP+ injury group,(D)sulforaphane pretreatment + MPP+ injury group.Cell viability was detected by MTT assay,and the sulforaphane pretreatment PC12 cell viability was observed in different concentrations.Flow cytometry was used to detect changes in the rate of apoptosis in different packet PC12 cells,and protein expression levels of nuclear factor erythroid2-related factor 2 (Nrf2),heme oxygenase (HO-1) and human NAD (P) H dehydrogenase,quinone 1 (NQO1) were detected by Western blot when the PC12 cells were incubated with sulforaphane (2.5 μmol/L) and (or) MPP+ (500 μmol/L) for 24 h in vitro.Results Compared to control group (cell survival rate was 98.70％),the survival percents of PC12 cells were significantly decreased in MPP+-treated group (58.16％).A significant difference was showed between group A and C (F =21.83,P ＜ 0.05),and the cell survival rate in group D was significantly improved.Compared to control group,the rate of apoptosis in MPP+ injury group was increased,and the rate of apoptosis after pretreatment of the sulforaphane was significantly reduced.Compared to MPP+ injury group,the levels of Nrf2,HO-1 and NQO1 protein expression were significantly increased in sulforaphane pretreatment group.Conclusion Sulforaphane have a protective effect against MPP+-induced PC12 cell model damage,and the protective effect may be achieved by activating the Nrf2-antioxidant response element pathway.
