Nutrition deficiency promotes apoptosis of cartilage endplate stem cells in a caspase-independent manner partially through upregulating BNIP3
Nutrition deficiency promotes apoptosis of cartilage endplate stem cells in a caspase-independent manner partially through upregulating BNIP3
复制标题
营养缺乏部分通过上调 BNIP3 以不依赖 caspase 的方式促进软骨终板干细胞凋亡
DOI：
10.1093/abbs/gmw111
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发表时间：
2017-01-01
影响因子：
3.7
通讯作者：
Wang, Jian
中科院分区：
文献类型：
--
作者：
He, Zhiliang;Pu, Luqiao;Wang, Jian
营养缺乏可导致兔软骨细胞凋亡和软骨终板(CEP)变性。软骨终板干细胞对软骨终板干细胞的结构和功能的完整性具有重要意义。BCL-2/腺病毒E1B19-kDa相互作用蛋白3(BNIP3)被报道调节细胞凋亡、自噬和细胞保护。在本研究中，我们旨在确定营养缺乏是否诱导CESCs的凋亡，以及BNIP3相关途径在营养缺乏时是否在CESCs中被激活。从变性的人CEP中分离出CESCs，在正常和营养缺乏的条件下进行培养。用流式细胞仪检测细胞凋亡率。采用实时定量聚合酶链式反应、Western印迹分析和免疫荧光法分别检测BNIP3的表达和细胞内定位。用JC-1染色和caspase-3活性检测线粒体膜电位和caspase-3活性。我们的结果表明营养缺乏促进了CESCs的凋亡和BNIP3的表达。值得注意的是，BNIP3基因的敲除可以部分减少营养缺乏诱导的CESCs的凋亡。此外，营养缺乏还可诱导CESCs中BNIP3的表达上调，导致BNIP3的线粒体易位和基质金属蛋白酶的丢失，并呈时间依赖性。然而，营养缺乏对CESCs的caspase-3活性没有影响。综上所述，营养缺乏可能部分通过上调BNIP3来促进CESC的凋亡，这可能导致BNIP3相关途径的激活，并以caspase不依赖的方式诱导CESCs的凋亡。
Nutrition deficiency is reported to induce apoptosis of chondrocytes and degeneration of cartilage endplate (CEP) in rabbit. Cartilage endplate stem cells (CESCs) are important for the integrity of structure and function of CEP. Bcl-2/adenovirus E1B 19-kDa-interacting protein 3 (BNIP3) has been reported to regulate apoptosis, autophagy, and cytoprotection. In this study, we aimed to determine whether nutrition deficiency induces apoptosis of CESCs, and whether or not the BNIP3-related pathway is activated in CESCs during nutrition deficiency. CESCs isolated from degenerated human CEP were cultured under normal or nutrition-deficient condition. Then, apoptosis was analyzed by flow cytometry. The expression and intracellular localization of BNIP3 were detected by quantitative real-time polymerase chain reaction, western blot analysis, and immunofluorescence assay, respectively. Mitochondrial membrane potential (MMP) and caspase-3 activity were measured by JC-1 staining and caspase-3 activity assay. Our results showed that nutrition deficiency promotes apoptosis and BNIP3 expression in CESCs. Notably, knockdown of BNIP3 could partially decrease nutrition deficiency-induced apoptosis of CESCs. In addition, nutrition deficiency could also induce upregulation of BNIP3, resulting in mitochondrial translocation of BNIP3 and loss of MMP in CESCs in a time-dependent manner. However, nutrition deficiency showed no effects on caspase-3 activity in CESCs. In summary, nutrition deficiency may promote CESC apoptosis partially through upregulating BNIP3, which might lead to activation of the BNIP3-related pathway and apoptosis of CESCs in a caspase-independent manner.