紫外诱导刺参自溶过程中细胞凋亡现象的研究
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- 英文论文题名：Analysis of apoptosis in ultraviolet-induced sea cucumber(Stichopus Japonicus) melting
- 论文作者：衣萌 ; 李鹏飞 ; 杨静峰
- 英文论文作者：Yi Meng ; Li Pengfei ; Yang Jingfeng ; School offoodscience and technology ; Dalian Polytechnic University ; National Engineering Research Center of Seafood
- 年：2016
- 作者机构：大连工业大学食品学院;国家海洋食品工程技术研究中心;
- 论文关键词：细胞凋亡 ; 自溶 ; TUNEL ; caspase-3
- 英文论文关键词：apoptosis ; melting ; TUNEL ; caspase-3
- 会议召开时间：2016-11-09
- 会议录名称：中国食品科学技术学会第十三届年会论文摘要集
- 英文会议录名称：Abstracts of the 13th Annual Meeting of CIFST
- 语种：中文
- 分类号：TS254.4
- 学会代码：ZGSP
- 会议名称：中国食品科学技术学会第十三届年会
- 会议地点：中国北京
- 主办单位：中国食品科学技术学会
- 学会名称：中国食品科学技术学会
- 页数：2
- 文件大小：106k
- 原文格式：O
- 会议级别：全国
摘要
刺参是一种营养及药用价值极高的海洋棘皮类水产食品。但是刺参具有极强的自溶能力,其分子机制至今仍然不清楚。新鲜刺参在紫外光下以0.056mW/cm~2的光强度下照射0.5小时引发自溶,于室温下静置6小时,采用TUNEL法研究刺参体壁细胞DNA片段化现象、以免疫组织切片技术Caspase-3进行免疫标记。同时检测了Caspase-9和Caspase-3的酶活变化情况。采用western blot技术对细胞色素C的迁移现象进行了研究。TUNEL实验结果表明,与新鲜刺参对照组相比,经过紫外诱导的刺参体壁细胞从0小时至6小时发生了显著的DNA片段化,在诱导后6小时时体壁中超过50%的细胞发生了DNA片段化,DNA片段化是细胞凋亡发生的标志性事件;紫外诱导后Caspase-3酶发生了极显著的升高,Caspase-3是细胞凋亡的效应分子,一旦被激活即发生不可逆性凋亡,也是细胞凋亡发生的标志性事件;采用western blot技术对细胞色素C的研究表明,线粒体外细胞质中的细胞色素C含量随自溶时间的延长发生了显著的升高。Caspase-9和Caspase-3的比活力也发生了显著升高,这表明,刺参细胞凋亡是通过线粒体凋亡途径实现的。
Sea cucumber(Stichopus japonicus) belongs to echinoderms and it is a kind of marine aquatic food with high nutritional and medicinal value.However,it shows strong ability of melting under severe case,and the mechanism of this phenomenon remains unclear.Fresh sea cucumbers were exposed to UV radiation for half an hour at an intensity of 0.056mW/cm and then held at room temperature for 6h to induce melting.The samples were analyzed with terminal deoxynucleotidyl-transferasemediated dUTP nick end-labeling(TUNEL) assay to detect the DNA fragmentation.Cleaved Caspase-3 was also detected with Immunohistochemistry method.The activity changes of Caspase-9 and Caspase-3 was detected too.Western-blot was performed to study the release of Cytochrome C transferfrom the inter-mitochondrial membrane space into the cytosol.The result of TUNEL indicated that the DNA fragmentation phenomenon was gradually grow from 0 to 6h compared with the fresh sea cucumber control group after exposuring to UV radiation.The body wall cells occurs DNA fragmentation phenomenon increased to greater than 50%after 6h exposure to UV radiation.The DNA fragmentation was the hallmark of apoptosis.The enzyme activity of Caspase-3 significandy increased after UV irradiation.Caspase-3 was the effector of apoptosis,which once activated irreversibly apoptosis will occur.It is also the hallmark of apoptosis.The determination of Cytochrome C indicates that the content of Cytochrome C outside mitochondria significantly increased with melting time goes on.The specific activity of Caspase-9 and Caspase-3 significantly increased.The results indicate that the apoptosis of sea cucumber was achieved by mitochondrial pathway.
Sea cucumber(Stichopus japonicus) belongs to echinoderms and it is a kind of marine aquatic food with high nutritional and medicinal value.However,it shows strong ability of melting under severe case,and the mechanism of this phenomenon remains unclear.Fresh sea cucumbers were exposed to UV radiation for half an hour at an intensity of 0.056mW/cm and then held at room temperature for 6h to induce melting.The samples were analyzed with terminal deoxynucleotidyl-transferasemediated dUTP nick end-labeling(TUNEL) assay to detect the DNA fragmentation.Cleaved Caspase-3 was also detected with Immunohistochemistry method.The activity changes of Caspase-9 and Caspase-3 was detected too.Western-blot was performed to study the release of Cytochrome C transferfrom the inter-mitochondrial membrane space into the cytosol.The result of TUNEL indicated that the DNA fragmentation phenomenon was gradually grow from 0 to 6h compared with the fresh sea cucumber control group after exposuring to UV radiation.The body wall cells occurs DNA fragmentation phenomenon increased to greater than 50%after 6h exposure to UV radiation.The DNA fragmentation was the hallmark of apoptosis.The enzyme activity of Caspase-3 significandy increased after UV irradiation.Caspase-3 was the effector of apoptosis,which once activated irreversibly apoptosis will occur.It is also the hallmark of apoptosis.The determination of Cytochrome C indicates that the content of Cytochrome C outside mitochondria significantly increased with melting time goes on.The specific activity of Caspase-9 and Caspase-3 significantly increased.The results indicate that the apoptosis of sea cucumber was achieved by mitochondrial pathway.
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