Inhibition of HSP90 protects cultured neurons from oxygen-glucose deprivation induced necroptosis by decreasing RIP3 expression
Inhibition of HSP90 protects cultured neurons from oxygen-glucose deprivation induced necroptosis by decreasing RIP3 expression
复制标题
HSP90α 的抑制通过降低 RIP3 表达来保护培养的神经元免受缺氧葡萄糖诱导的坏死性凋亡
DOI：
10.1002/jcp.26294
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发表时间：
2018-06-01
影响因子：
5.6
通讯作者：
Xiong, Kun
中科院分区：
文献类型：
--
作者：
Wang, Zhen;Guo, Li-min;Xiong, Kun
热休克蛋白90 （HSP90）分别维持细胞稳定和调节细胞死亡。最近的研究表明，HSP90参与了受体相互作用蛋白3 （RIP3）介导的HT29细胞坏死坏死。已知氧和葡萄糖剥夺（OGD）可诱导坏死下垂，这是由RIP3在神经元中调节的。然而，HSP90是否通过调控RIP3参与ogd诱导的培养神经元坏死下垂过程尚不清楚。我们的研究发现，在暴露于OGD损伤后，原代培养的皮质神经元和PC-12细胞发生坏死性下垂。此外，通过免疫荧光（IF）染色、免疫印迹（WB）和免疫沉淀（IP）测定，RIP3/p-RIP3、MLKL/p-MLKL和RIP1/RIP3复合物（坏死体）的表达在OGD后显著增加。此外，计算机模拟和IP分析的数据表明，HSP90与RIP3相互作用。此外，通过WB和IF染色，在培养的神经元中，HSP90在OGD后过表达。使用特异性抑制剂格尔达霉素（GA）和HSP90的siRNA/shRNA抑制培养神经元中的HSP90，可保护培养神经元免于坏死。我们的研究表明，HSP90抑制剂GA不仅通过降低总RIP3/MLKL的表达，而且通过降低p-RIP3/p-MLKL和RIP1/RIP3坏死体的表达来拯救培养的神经元。在这项研究中，我们发现抑制HSP90可以通过调节RIP3的表达来保护原代培养的皮质神经元和PC-12细胞免受ogd诱导的坏死性凋亡。
Heat shock protein 90 (HSP90) maintains cell stabilization and regulates cell death, respectively. Recent studies have shown that HSP90 is involved in receptor interacting protein 3 (RIP3)-mediated necroptosis in HT29 cells. It is known that oxygen and glucose deprivation (OGD) can induce necroptosis, which is regulated by RIP3 in neurons. However, it is still unclear whether HSP90 participates in the process of OGD-induced necroptosis in cultured neurons via the regulation of RIP3. Our study found that necroptosis occurs in primary cultured cortical neurons and PC-12 cells following exposure to OGD insult. Additionally, the expression of RIP3/p-RIP3, MLKL/p-MLKL, and the RIP1/RIP3 complex (necrosome) significantly increased following OGD, as measured through immunofluorescence (IF) staining, Western blotting (WB), and immunoprecipitation (IP) assay. Additionally, data from computer simulations and IP assays showed that HSP90 interacts with RIP3. In addition, HSP90 was overexpressed following OGD in cultured neurons, as measured through WB and IF staining. Inhibition of HSP90 in cultured neurons, using the specific inhibitor, geldanamycin (GA), and siRNA/shRNA of HSP90, protected cultured neurons from necrosis. Our study showed that the inhibitor of HSP90, GA, rescued cultured neurons not only by decreasing the expression of total RIP3/MLKL, but also by decreasing the expression of p-RIP3/p-MLKL and the RIP1/RIP3 necrosome. In this study, we reveal that inhibition of HSP90 protects primary cultured cortical neurons and PC-12 cells from OGD-induced necroptosis through the modulation of RIP3 expression.