目的:探讨白藜芦醇(Res)对热刺激诱导的小鼠心肌细胞(MCM)凋亡的保护作用。方法:复制热刺激诱导MCM凋亡模型。首先采用实时无标记细胞分析技术检测Res对MCM存活率的影响;然后采用Annexin V-FITC/7-AAD双染法检测不同浓度Res给药组对MCM...目的:探讨白藜芦醇(Res)对热刺激诱导的小鼠心肌细胞(MCM)凋亡的保护作用。方法:复制热刺激诱导MCM凋亡模型。首先采用实时无标记细胞分析技术检测Res对MCM存活率的影响;然后采用Annexin V-FITC/7-AAD双染法检测不同浓度Res给药组对MCM凋亡率的影响。JC-1探针法检测线粒体膜电位,初步确证MCM凋亡依赖于线粒体凋亡途径。利用全自动荧光酶标仪检测荧光探针(Ac-LEHD)2-R110标记的Caspase-9相对表达量;利用流式细胞仪检测荧光探针TF2-DEVD-FMK标记的Caspase-3/7的相对表达量;Western Blot法检测Bax、Bcl-2、COXⅣ蛋白的表达;ELISA方法检测细胞色素C(Cyt-C)含量、比色法测定Cyt-C氧化酶活性。结果:与模型组比较,Res高剂量及中剂量组细胞凋亡率显著降低(P<0.05,P<0.01),各组均可显著改善线粒体膜电位(P<0.01),Res高剂量及低剂量组Cyt-C释放量显著降低(P<0.05,P<0.01),Res各组均可显著降低Caspase-9活性(P<0.01),R es高剂量组可显著降低Ca s p a s e-3/7活性(P<0.01),B c l-2/B ax比值显著升高(P<0.01,P<0.05),Res干预后Cyt-C氧化酶活性均升高但差异无统计学意义。结论:Res通过抑制线粒体凋亡途径缓解热刺激诱导的MCM凋亡。展开更多
Disruption of mitochondrial reactive oxygen species （mitoROS） plays a major role in cancer cell apoptosis. Here, we designed a core/shell-structured mitochondriatargeting upconversion-based nano-photosensitizer （TP...Disruption of mitochondrial reactive oxygen species （mitoROS） plays a major role in cancer cell apoptosis. Here, we designed a core/shell-structured mitochondriatargeting upconversion-based nano-photosensitizer （TPP-UC（PS）） with a lanthanidedoped upconversion nanoparticle （UCNP） core coated by a photosensitizer （PS）-incorporated dense silica shell. Following irradiation with external nearinfrared laser （NIR）, TPP-UC（PS） in mitochondria caused serious mitochondrial matrix swelling for the activated upconversion-based photodynamic therapy （UC-PDT）, and the mobilization of cytochrome c （cyt c） was amplified in response to the increased mitoROS. Specifically, this heme-containing cyt c could be monitored by varying TPP-UC（PS）＇s upconversion luminescence signal （UCL）, which may facilitate the in situ detection of cyt c for apoptosis research. As a proof of concept, our designed TPP-UC（PS） may provide significant opportunities for controlling cancer cell apoptosis under NIR stimulation and for studying apoptosis using the dynamic UCL, which is influenced by local cyt c.展开更多
文摘目的:探讨白藜芦醇(Res)对热刺激诱导的小鼠心肌细胞(MCM)凋亡的保护作用。方法:复制热刺激诱导MCM凋亡模型。首先采用实时无标记细胞分析技术检测Res对MCM存活率的影响;然后采用Annexin V-FITC/7-AAD双染法检测不同浓度Res给药组对MCM凋亡率的影响。JC-1探针法检测线粒体膜电位,初步确证MCM凋亡依赖于线粒体凋亡途径。利用全自动荧光酶标仪检测荧光探针(Ac-LEHD)2-R110标记的Caspase-9相对表达量;利用流式细胞仪检测荧光探针TF2-DEVD-FMK标记的Caspase-3/7的相对表达量;Western Blot法检测Bax、Bcl-2、COXⅣ蛋白的表达;ELISA方法检测细胞色素C(Cyt-C)含量、比色法测定Cyt-C氧化酶活性。结果:与模型组比较,Res高剂量及中剂量组细胞凋亡率显著降低(P<0.05,P<0.01),各组均可显著改善线粒体膜电位(P<0.01),Res高剂量及低剂量组Cyt-C释放量显著降低(P<0.05,P<0.01),Res各组均可显著降低Caspase-9活性(P<0.01),R es高剂量组可显著降低Ca s p a s e-3/7活性(P<0.01),B c l-2/B ax比值显著升高(P<0.01,P<0.05),Res干预后Cyt-C氧化酶活性均升高但差异无统计学意义。结论:Res通过抑制线粒体凋亡途径缓解热刺激诱导的MCM凋亡。
基金This work has been financially supported by the National Natural Science Foundation of China （Nos. 51372260, 51132009, and 81471714）, and the Shanghai Excellent Academic Leaders Program （No. 16XD1404000）. Thanks to Linlin Zhang, Heliang Yao, Qingfeng Xiao,Huaiyong Xing, Wenpei Fan, Zhaowen Cui, Li Jiang, and Jianan Liu from Shanghai Institute of Ceramics, Chinese Academy of Sciences for useful discussions.
文摘Disruption of mitochondrial reactive oxygen species （mitoROS） plays a major role in cancer cell apoptosis. Here, we designed a core/shell-structured mitochondriatargeting upconversion-based nano-photosensitizer （TPP-UC（PS）） with a lanthanidedoped upconversion nanoparticle （UCNP） core coated by a photosensitizer （PS）-incorporated dense silica shell. Following irradiation with external nearinfrared laser （NIR）, TPP-UC（PS） in mitochondria caused serious mitochondrial matrix swelling for the activated upconversion-based photodynamic therapy （UC-PDT）, and the mobilization of cytochrome c （cyt c） was amplified in response to the increased mitoROS. Specifically, this heme-containing cyt c could be monitored by varying TPP-UC（PS）＇s upconversion luminescence signal （UCL）, which may facilitate the in situ detection of cyt c for apoptosis research. As a proof of concept, our designed TPP-UC（PS） may provide significant opportunities for controlling cancer cell apoptosis under NIR stimulation and for studying apoptosis using the dynamic UCL, which is influenced by local cyt c.
