摘要
目的 :设计、合成Caspase 3mRNA反义寡核苷酸 ,并筛选出能阻抑Aβ2 5～ 3 5诱导的PC 12细胞凋亡的有效片段。方法 :针对Caspase 3mRNA不同区域设计出一系列 17～ 18 mer反义寡核苷酸 ,用脂质体导入PC 12细胞内 ,并用Aβ2 5～ 3 5诱导细胞凋亡。用AnnexinⅤ FITC/PI复染、流式细胞术检测观察凋亡早期细胞变化情况。结果 :针对Caspase 3mRNA编码起始区的反义寡核苷酸片段 (ASODN2 ,-6～ 12 ,5’ GTTGTTGTCCATGGTCAC 3’)导入PC 12细胞后 ,在Aβ2 5～ 3 5诱导下 ,AnnexinⅤ /PI复染、流式细胞仪检测见早期凋亡细胞较对照组及其它反义寡核苷酸组明显减少 (P <0 .0 5 ) ;且导入后Aβ2 5～ 3 5诱导凋亡细胞数与无诱导组相比未见明显增多 (P >0 .0 5 )。结论 :1.Caspase 3在Aβ2 5～ 3 5诱导的PC 12细胞凋亡中起重要作用 ;2 .Caspase 3mRNA的反义寡核苷酸片段 5’ GTTGTTGTCCATGGTCAC 3’能有效阻止Aβ2 5～ 3 5诱导的PC
Objective: To design and synthesize antisense oligodexynucleotides (ASODNs) targeting Caspase-3 mRNA, and to screen the effective ASODNs to prevent PC-12 cells apoptosis induced by Aβ 25～35. Methods: 17～18-mer ASODNs targeting different regions of Caspase-3 mRNA were designed and transfected into PC-12 cells by Lipofectimine followed by induction with Aβ 25～35. The effective ASOSNs preventing PC-12 cells apoptosis were screened according to the results of Flow Cytometry (FCM) analysis with Annexin V-FITC/Propidium Iodie (PI) dual staining. Results: FCM analysis with Annexin V/PI staining showed that the ratio of apoptotic cells decreased significantly than other ODNS after the ASODN 2 (5'-GTT GTT GTC CAT GGT CAC-3'), which targets bases - 6～12 of Caspase-3 mRNA, were transfected into PC-12 cells and then cells were expose to Aβ 25～35. Apoptotic cells transfected with ASODN 2 treatment with Aβ 25～35 were non-differentiable statistically to Aβ 25～35 group. Conclusions: Caspase-3 plays an important role in the PC-12 cells apoptosis treatment with Aβ 25～35. The ASODN (5'-GTT GTT GTC CAT GGT CAC-3') aimed at Caspase-3 mRNA is identified as an effective fragment to prevent the PC-12 cells apoptosis treatment with Aβ 25～35.
出处
《中国临床解剖学杂志》
CSCD
北大核心
2003年第6期617-619,626,共4页
Chinese Journal of Clinical Anatomy
基金
国家自然科学基金 (39880 0 0 8)
陕西省自然科学基金(98sm61 )资助项目
