Wang Shixuan, Kwon Sang-Ho, Su Yunchao, Dong Zheng
Department of Cellular Biology and Anatomy, Medical College of Georgia, Augusta University , Augusta, Georgia.
Department of Pharmacology and Toxicology, Medical College of Georgia, Augusta University, and Charlie Norwood Veterans Affairs Medical Center , Augusta, Georgia.
Am J Physiol Renal Physiol. 2019 Jul 1;317(1):F116-F123. doi: 10.1152/ajprenal.00139.2019. Epub 2019 May 15.
Stress granules (SGs) are a type of cytoplasmic structures formed in eukaryotic cells upon cell stress, which mainly contain RNA-binding proteins and RNAs. The formation of SGs is generally regarded as a mechanism for cells to survive a harsh insult. However, little is known about SG formation and function in kidneys. To address this, we applied different kinds of stressors to cultured proximal tubular cells as well as a short period of ischemia-reperfusion to mouse kidneys. It was found that glycolytic inhibitors such as 2-deoxy-d-glucose and 3-(3-pyridinyl)-1-(4-pyridinyl)-2-propen-1-one induced SG formation within 30 min in these cells. Similarly, SGs were induced by inhibitors of mitochondrial respiration such as sodium azide and CCCP. Renal ischemia-reperfusion induced SG formation in the cells of proximal tubules. To test the role of SGs, we stably knocked down G3bp1, a SG core protein, in renal tubular cells by shRNA viral transduction. As expected, knockdown of G3bp1 largely disrupted the assembly of SGs. After azide or cisplatin treatment, more dead cells were found in knockdown cells compared with controls, accompanied by increases in cleaved/active caspase-3. Reintroduction of exogenous G3bp1 into knockdown cells could rescue the cell death phenotype. Taken together, our data provide the first evidence of SG formation in renal tubular cells during metabolic stress and acute kidney injury. SGs are formed to protect proximal tubular cells under these conditions. Modulation of SG biogenesis may provide a novel approach to lessen the severity of renal diseases.
应激颗粒(SGs)是真核细胞在受到细胞应激时形成的一种细胞质结构,主要包含RNA结合蛋白和RNA。SGs的形成通常被认为是细胞在遭受严重损伤时存活的一种机制。然而,关于SGs在肾脏中的形成和功能知之甚少。为了解决这个问题,我们对培养的近端肾小管细胞施加了不同类型的应激源,并对小鼠肾脏进行了短时间的缺血再灌注。结果发现,糖酵解抑制剂如2-脱氧-D-葡萄糖和3-(3-吡啶基)-1-(4-吡啶基)-2-丙烯-1-酮在30分钟内就能诱导这些细胞形成SGs。同样,线粒体呼吸抑制剂如叠氮化钠和CCCP也能诱导SGs形成。肾脏缺血再灌注可诱导近端小管细胞形成SGs。为了测试SGs的作用,我们通过shRNA病毒转导在肾小管细胞中稳定敲低了SG核心蛋白G3bp1。正如预期的那样,G3bp1的敲低极大地破坏了SGs的组装。在叠氮化钠或顺铂处理后,与对照组相比,敲低细胞中发现更多的死亡细胞,同时裂解/活性半胱天冬酶-3增加。将外源性G3bp1重新引入敲低细胞中可以挽救细胞死亡表型。综上所述,我们的数据首次证明了在代谢应激和急性肾损伤期间肾小管细胞中SGs的形成。在这些条件下,SGs的形成是为了保护近端肾小管细胞。调节SG生物发生可能为减轻肾脏疾病的严重程度提供一种新方法。