Critical contribution of RIPK1 mediated mitochondrial dysfunction and oxidative stress to compression-induced rat nucleus pulposus cells necroptosis and apoptosis
Critical contribution of RIPK1 mediated mitochondrial dysfunction and oxidative stress to compression-induced rat nucleus pulposus cells necroptosis and apoptosis
复制标题
RIPK1介导的线粒体功能障碍和氧化应激对压迫诱导的大鼠髓核细胞坏死性凋亡和凋亡的关键作用
DOI：
10.1007/s10495-018-1455-x
复制
发表时间：
2018-06-01
期刊：
影响因子：
7.2
通讯作者：
Shao, Zengwu
中科院分区：
文献类型：
--
作者：
Chen, Songfeng;Lv, Xiao;Shao, Zengwu
本研究的目的是探讨RIPK 1介导的线粒体功能障碍和氧化应激是否有助于压力诱导的髓核细胞（NP）坏死性凋亡和凋亡，以及坏死性凋亡和凋亡之间的相互作用关系。大鼠NP细胞经历了不同时期的1.0 MPa压缩。为了确定压缩是否影响线粒体功能，我们评估了线粒体膜电位，线粒体通透性转换孔（mPTP），线粒体超微结构和ATP含量。氧化应激相关指标活性氧、超氧化物歧化酶和丙二醛也进行了评估。为了验证氧化应激与坏死性凋亡以及细胞凋亡之间的相关性，利用RIPK 1抑制剂necrostatin-1（Nec-1）、mPTP抑制剂环孢素A（CsA）、抗氧化剂和小干扰RNA技术。结果确定，压缩引起时间依赖性线粒体功能障碍和氧化应激升高。Nec-1和CsA恢复线粒体功能，减少氧化应激，这对应于减少坏死性凋亡和细胞凋亡。CsA下调线粒体亲环素D的表达，但对RIPK 1的表达和pRIPK 1的激活影响不大。此外，我们发现Nec-1在很大程度上阻断了细胞凋亡;而细胞凋亡抑制剂Z-VAD-FMK增加了RIPK 1的表达和pRIPK 1的激活，并且协调调节坏死性凋亡和细胞凋亡使NP细胞更有效地存活。与Nec-1相反，SiRIPK 1加剧了线粒体功能障碍和氧化应激。综上所述，RIPK 1介导的线粒体功能障碍和氧化应激在压迫性损伤时NP细胞坏死性凋亡和凋亡中起着至关重要的作用。坏死性凋亡和凋亡的协同调节可能对髓核细胞的存活产生更有利的影响，并最终延缓甚至延缓椎间盘退变。
The aim of this study was to investigate whether RIPK1 mediated mitochondrial dysfunction and oxidative stress contributed to compression-induced nucleus pulposus (NP) cells necroptosis and apoptosis, together with the interplay relationship between necroptosis and apoptosis in vitro. Rat NP cells underwent various periods of 1.0 MPa compression. To determine whether compression affected mitochondrial function, we evaluated the mitochondrial membrane potential, mitochondrial permeability transition pore (mPTP), mitochondrial ultrastructure and ATP content. Oxidative stress-related indicators reactive oxygen species, superoxide dismutase and malondialdehyde were also assessed. To verify the relevance between oxidative stress and necroptosis together with apoptosis, RIPK1 inhibitor necrostatin-1(Nec-1), mPTP inhibitor cyclosporine A (CsA), antioxidants and small interfering RNA technology were utilized. The results established that compression elicited a time-dependent mitochondrial dysfunction and elevated oxidative stress. Nec-1 and CsA restored mitochondrial function and reduced oxidative stress, which corresponded to decreased necroptosis and apoptosis. CsA down-regulated mitochondrial cyclophilin D expression, but had little effects on RIPK1 expression and pRIPK1 activation. Additionally, we found that Nec-1 largely blocked apoptosis; whereas, the apoptosis inhibitor Z-VAD-FMK increased RIPK1 expression and pRIPK1 activation, and coordinated regulation of necroptosis and apoptosis enabled NP cells survival more efficiently. In contrast to Nec-1, SiRIPK1 exacerbated mitochondrial dysfunction and oxidative stress. In summary, RIPK1-mediated mitochondrial dysfunction and oxidative stress play a crucial role in NP cells necroptosis and apoptosis during compression injury. The synergistic regulation of necroptosis and apoptosis may exert more beneficial effects on NP cells survival, and ultimately delaying or even retarding intervertebral disc degeneration.