摘要
目的 :探讨一氧化氮 (NO)和半胱氨酸蛋白酶 3 (caspase - 3 )在多巴胺诱导PC12细胞凋亡中的可能作用。方法 :流式细胞仪定量检测PC12细胞的凋亡率 ,原位末端标记法 (TUNEL)观察凋亡细胞的形态 ,Griess法测定NO-2 的浓度 ,荧光分光光度计法检测caspase - 3的活力 ,半定量RT -PCR法检测诱导型一氧化氮合酶 (iNOS)mRNA的表达水平。结果 :多巴胺 ( 0 .15 - 0 60mmol/L)可剂量依赖性地诱导PC12细胞凋亡 ,表现为凋亡细胞的TUNEL染色阳性 ;iNOSmRNA的表达、NO的合成及caspase - 3的活力均有明显的增加 (P <0 .0 1) ;特异性的iNOS抑制剂aminoguanidine和caspase - 3抑制剂Ac -DEVD -CHO通过减少NO的生成和抑制caspase- 3的激活阻断PC12细胞凋亡。结论 :NO的生成可能是多巴胺诱导PC12细胞凋亡的触发因子 ,caspase- 3的激活是其中的效应因子。
AIM: To investigate the effects of nitric oxide(NO) and caspase-3 on dopamine-induced apoptosis in PC12 cells. METHODS: Flow cytometric assay was used to quantify the apoptotic cells. The morphological of apoptotic cells was evaluated by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL). Nitrite was quantified by Griess reaction. Inducible nitric oxide synthase(iNOS) mRNA was identified by semiquantitative reverse transcription polymerase chain reaction(RT-PCR). Caspase-3 activity was determined by fluorescent spectrofluorometer. RESULTS: Dopamine induced PC12 cells apoptosis in a concentration-dependent manner (0.15-0.60 mmol/L), with positive TUNEL staining. During the development of apoptosis, the expression of iNOS mRNA and the levels of NO increased markedly, so did caspase-3 activity(P<0.01). Specific inhibitor of iNOS aminoguanidine and inhibitor of caspase-3 Ac-DEVD-CHO blocked PC12 cells apoptosis by decreasing the production of NO and inhibiting the activation of caspase-3. CONCLUSION:NO might be a trigger and caspase-3 might be an executor during dopamine induced apoptosis in PC12 cells.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2001年第10期971-975,共5页
Chinese Journal of Pathophysiology
基金
福建省自然科学基金重点课题资助(No.C982 0 0 4 )
