The RIP1-RIP3 Complex Mediates Osteocyte Necroptosis after Ovariectomy in Rats.
The RIP1-RIP3 Complex Mediates Osteocyte Necroptosis after Ovariectomy in Rats.
复制标题
DOI：
10.1371/journal.pone.0150805
复制
发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Jiang D
中科院分区：
文献类型：
--
作者：
Cui H;Zhu Y;Jiang D
据报道，骨细胞凋亡在骨重建中起着核心作用。除细胞凋亡外，骨细胞丢失还可能涉及其他机制。本研究旨在探讨坏死性下垂对去卵巢(OVX)大鼠骨细胞的影响。雌性SD大鼠96只，随机分为去卵巢组和假手术组。分别于术后0、4、8、12周取材，每组12例。测量骨密度(BMD)和体重。用透射电子显微镜和显微CT观察雌激素缺乏引起的细胞形态和骨微结构的变化。通过原位末端标记法和免疫荧光技术检测caspase-3活性，观察骨细胞的凋亡和坏死性下垂。卵巢切除后8周，大量具有典型坏死形态特征的骨细胞呈TUNEL阳性，而活性caspase-3呈阴性。Western blotting、实时定量聚合酶链式反应和免疫荧光检测显示，去卵巢后8周，骨细胞中受体相互作用的丝氨酸/苏氨酸蛋白激酶1(RIP1)和RIP3的水平显著升高。这些数据首次表明，在雌激素缺乏引起的骨质疏松的条件下，坏死性下垂加速了骨细胞的丢失。这些发现提供了骨细胞坏死性下垂与绝经后骨质疏松症相关的潜在机制的证据。
Osteocyte apoptosis has been reported to play a central role in bone remodeling. In addition to apoptosis, other mechanisms may be involved in osteocyte loss. This study aimed to investigate the effect of necroptosis on osteocytes in ovariectomized (OVX) rats. Ninety-six female Sprague-Dawley rats were randomly divided into an OVX group and a sham group. At 0, 4, 8 and 12 weeks after surgery, specimens from each group (n = 12 each) were harvested. Bone mineral density (BMD) and body weight were measured. Transmission electron microscopy (TEM) and micro-CT were used to observe the changes in cellular morphology and bone microarchitecture induced by estrogen deficiency. Osteocyte apoptosis and necroptosis were evaluated via TUNEL and immunofluorescence staining for active caspase-3. At 8 weeks after ovariectomy, a greater number of osteocytes with typical necrotic morphological features were TUNEL positive but negative for active caspase-3. Western blotting, quantitative real-time PCR and immunofluorescence assessments demonstrated that the levels of receptor-interacting serine/threonine protein kinase 1 (RIP1) and RIP3 in osteocytes were significantly increased at 8 weeks after ovariectomy. These data are the first to suggest that necroptosis accelerates osteocyte loss under conditions of estrogen deficiency-induced osteoporosis in OVX rats. These findings provide evidence of a potential mechanism through which osteocyte necroptosis is associated with postmenopausal osteoporosis.