Receptor interacting protein 3-induced RGC-5 cell necroptosis following oxygen glucose deprivation.
Receptor interacting protein 3-induced RGC-5 cell necroptosis following oxygen glucose deprivation.
复制标题
氧糖剥夺后受体相互作用蛋白 3 诱导 RGC-5 细胞坏死性凋亡
DOI：
10.1186/s12868-015-0187-x
复制
发表时间：
2015-08-04
期刊：
影响因子：
2.4
通讯作者：
Xiong K
中科院分区：
文献类型：
--
作者：
Ding W;Shang L;Huang JF;Li N;Chen D;Xue LX;Xiong K
坏死性凋亡是一种受调控的细胞死亡形式，与多种疾病的发病机制有关。受体相互作用蛋白3（Receptor-interacting protein 3，RIP 3）是RIP家族的一员，在心肌缺血、炎症性肠病、缺血性脑损伤等多种人类疾病中起重要的调控作用。我们之前的研究表明，RIP 3在大鼠视网膜神经节细胞（RGC）中表达，在急性高眼压的早期阶段显着上调。此外，RIP 3表达与碘化丙啶（PI）阳性染色（坏死细胞）共定位。提示RIP 3表达上调可能参与了损伤后RGCs的坏死过程。在这项研究中，我们的目的是揭示RIP 3可能参与氧葡萄糖剥夺（OGD）诱导的视网膜神经节细胞-5（RGC-5）坏死性凋亡。将RGC-5细胞在Dulbecco改良的基本培养基中培养，并通过8 h OGD诱导坏死性凋亡。PI染色和流式细胞术检测RGC-5坏死。Western blot检测RIP 3的表达，流式细胞术检测RIP 3对RGC-5凋亡的影响。丙二醛（MDA）脂质过氧化测定，以确定氧化应激的程度。PI染色显示OGD诱导的RGC-5细胞死亡的早期阶段存在坏死。使用坏死性凋亡抑制剂necrostatin-1通过流式细胞术检测OGD后RGC-5坏死性凋亡的存在。Western blot结果表明RIP 3表达上调可能参与了RGC-5的坏死性凋亡。流式细胞术显示，在rip 3敲低后，OGD诱导的坏死RGC-5细胞的数量减少。此外，正常RGC-5细胞中的MDA水平远高于OGD后rip 3敲低细胞中的MDA水平。我们的研究结果表明，RGC-5细胞坏死凋亡OGD后介导的RIP 3诱导的氧化应激增加。
Necroptosis is a type of regulated form of cell death that has been implicated in the pathogenesis of various diseases. Receptor-interacting protein 3 (RIP3), a member of the RIP family of proteins, has been reported as an important necroptotic pathway mediator in regulating a variety of human diseases, such as myocardial ischemia, inflammatory bowel disease, and ischemic brain injury. Our previous study showed that RIP3 was expressed in rat retinal ganglion cells (RGCs), where it was significantly upregulated during the early stage of acute high intraocular pressure. Furthermore, RIP3 expression was co-localized with propidium iodide (PI)-positive staining (necrotic cells). These results suggested that RIP3 up-regulation might be involved in the necrosis of injured RGCs. In this study, we aimed to reveal the possible involvement of RIP3 in oxygen glucose deprivation (OGD)-induced retinal ganglion cell-5 (RGC-5) necroptosis. RGC-5 cells were cultured in Dulbecco’s-modified essential medium and necroptosis was induced by 8 h OGD. PI staining and flow cytometry were performed to detect RGC-5 necrosis. RIP3 expression was detected by western blot and flow cytometry was used to detect the effect of RIP3 on RGC-5 necroptosis following OGD in rip3 knockdown cells. Malondialdehyde (MDA) lipid peroxidation assay was performed to determine the degree of oxidative stress. PI staining showed that necrosis was present in the early stage of OGD-induced RGC-5 cell death. The presence of RGC-5 necroptosis after OGD was detected by flow cytometry using necrostatin-1, a necroptosis inhibitor. Western blot demonstrated that RIP3 up-regulation may be involved in RGC-5 necroptosis. Flow cytometry revealed that the number of OGD-induced necrotic RGC-5 cells was reduced after rip3 knockdown. Furthermore, MDA levels in the normal RGC-5 cells were much higher than in the rip3-knockdown cells after OGD. Our findings suggest that RGC-5 cell necroptosis following OGD is mediated by a RIP3-induced increase in oxidative stress.