| Fas对镉激活PC12细胞线粒体凋亡通路的影响 |
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| 引用本文： | 闻双全,陈洁,王莉,邹辉,顾建红,刘学忠,卞建春,刘宗平,袁燕.Fas对镉激活PC12细胞线粒体凋亡通路的影响[J].畜牧兽医学报,2021,52(8):2326-2333. |
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| 作者姓名： | 闻双全 陈洁 王莉 邹辉 顾建红 刘学忠 卞建春 刘宗平 袁燕 |
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| 作者单位： | 1. 扬州大学兽医学院, 扬州 225009;2. 江苏高校动物重要疫病与人兽共患病防控协同创新中心, 扬州 225009;3. 江苏省南通市动物疫病预防控制中心, 南通 226011 |
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| 基金项目： | 国家自然科学基金资助项目（31772808）；江苏高校优势学科建设工程资助项目（PAPD）；扬州大学优秀青年骨干教师资助项目；扬州大学校教改课题（YZUJX2019-51C） |
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| 摘 要： | 旨在探究死亡受体Fas在镉致大鼠肾上腺嗜铬细胞瘤细胞（PC12）凋亡中的作用及其对线粒体通路的调控机制，用10 μmol·L-1镉处理Fas基因沉默的PC12细胞株12 h，通过Western blot检测BH3相互作用域死亡激动剂（BID）、半胱氨酸蛋白酶-9（caspase-9）、半胱氨酸蛋白酶-3（caspase-3）、多聚二磷酸腺苷核糖聚合酶（PARP）的活化情况，Bcl-2相关X蛋白（Bax）、B细胞淋巴瘤/白血病-2基因（Bcl-2）、凋亡诱导因子（AIF）、核酸内切酶G（Endo G）的表达情况，以及细胞色素C（Cyt C）在细胞内的分布情况，免疫荧光染色检测AIF核转位。结果显示，镉极显著上调tBID/BID比值和Bax/Bcl-2比值，诱导Cyt C从线粒体释放到细胞质，激活caspase-9、caspase-3和PARP，增加AIF和Endo G蛋白表达水平（P<0.01），并诱导AIF核转位；沉默Fas极显著抑制镉引起的tBID/BID比值和Bax/Bcl-2比值升高，Cyt C从线粒体释放到胞浆，caspase-3、PARP蛋白活化和AIF、Endo G蛋白表达水平极显著升高（P<0.01），显著抑制镉激活的caspase-9（P<0.05），并抑制AIF核转位。综上表明，Fas通过调控线粒体通路参与镉致PC12细胞凋亡。
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| 关 键 词： | 镉 Fas PC12细胞 凋亡 线粒体通路 |
| 收稿时间： | 2020-12-14 |
Effects of Fas on Mitochondrial Apoptotic Pathway Activated by Cadmium in PC12 Cells |
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| WEN Shuangquan,CHEN Jie,WANG Li,ZOU Hui,GU Jianhong,LIU Xuezhong,BIAN Jianchun,LIU Zongping,YUAN Yan.Effects of Fas on Mitochondrial Apoptotic Pathway Activated by Cadmium in PC12 Cells[J].Acta Veterinaria et Zootechnica Sinica,2021,52(8):2326-2333. |
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| Authors: | WEN Shuangquan CHEN Jie WANG Li ZOU Hui GU Jianhong LIU Xuezhong BIAN Jianchun LIU Zongping YUAN Yan |
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| Institution: | 1. College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China;2. Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China;3. Nantong Animal Disease Prevention and Control Center, Nantong 226011, China |
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| Abstract: | In order to explore the role of Fas death receptor in rat pheochromocytoma cell line (PC12) apoptosis induced by cadmium (Cd) and its regulation mechanism on the mitochondrial pathway, Fas gene silencing PC12 cells were treated with 10 μmol·L-1 Cd for 12 hours. The activation of BH3 interacting domain death agonist (BID), cysteine aspartate-specific protease-9 (caspase-9), cysteine aspartate-specific protease-3 (caspase-3), poly ADP ribose polymerase (PARP), the expression of Bcl-2 associated X protein (Bax), B-cell lymphoma/leukemia-2 (Bcl-2), apoptosis inducing factor (AIF), endonuclease G (Endo G), and the distribution of cytochrome C (Cyt C) in cells were detected by Western blot. Nuclear translocation of AIF was detected by immunofluorescence staining. The results showed that Cd significantly increased the tBID/BID and Bax/Bcl-2 ratios, the release of Cyt C from mitochondria into the cytosol, the activation of caspase-9, caspase-3, PARP, and the expression of AIF, Endo G (P<0.01). Meanwhile, Cd induced the nuclear translocation of AIF. Fas silencing significantly inhibited Cd-induced increase of the tBID/BID and Bax/Bcl-2 ratios, the release of Cyt C from mitochondria into the cytosol, the activation of caspase-3, PARP, the expression of AIF, Endo G (P<0.01). Cd-activated caspase-9 was significantly inhibited by Fas silencing (P<0.05). Cd-induced nuclear translocation of AIF was abated by Fas silencing. The above results indicate that PC12 cells apoptosis induced by Cd via the mitochondrial pathway is mediated by Fas. |
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| Keywords: | cadmium Fas PC12 cells apoptosis mitochondrial pathway |
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