Al-Bataineh Mohammad M, Kinlough Carol L, Poland Paul A, Pastor-Soler Núria M, Sutton Timothy A, Mang Henry E, Bastacky Sheldon I, Gendler Sandra J, Madsen Cathy S, Singh Sucha, Monga Satdarshan P, Hughey Rebecca P
Renal-Electrolyte Division, Department of Medicine, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania;
Division of Nephrology and Hypertension, Department of Medicine, University of Southern California/UKRO Kidney Research Center, Keck School of Medicine of the University of Southern California, Los Angeles, California;
Am J Physiol Renal Physiol. 2016 Mar 15;310(6):F569-79. doi: 10.1152/ajprenal.00520.2015. Epub 2016 Jan 6.
The hypoxia-inducible factor (HIF)-1 and β-catenin protective pathways represent the two most significant cellular responses that are activated in response to acute kidney injury. We previously reported that murine mucin (Muc)1 protects kidney function and morphology in a mouse model of ischemia-reperfusion injury (IRI) by stabilizing HIF-1α, enhancing HIF-1 downstream signaling, and thereby preventing metabolic stress (Pastor-Soler et al. Muc1 is protective during kidney ischemia-reperfusion injury. Am J Physiol Renal Physiol 308: F1452-F1462, 2015). We asked if Muc1 regulates the β-catenin protective pathway during IRI as 1) β-catenin nuclear targeting is MUC1 dependent in cultured human cells, 2) β-catenin is found in coimmunoprecipitates with human MUC1 in extracts of both cultured cells and tissues, and 3) MUC1 prevents β-catenin phosphorylation by glycogen synthase kinase (GSK)3β and thereby β-catenin degradation. Using the same mouse model of IRI, we found that levels of active GSK3β were significantly lower in kidneys of control mice compared with Muc1 knockout (KO) mice. Consequently, β-catenin was significantly upregulated at 24 and 72 h of recovery and appeared in the nuclear fraction at 72 h in control mouse kidneys. Both β-catenin induction and nuclear targeting were absent in Muc1 KO mice. We also found downstream induction of β-catenin prosurvival factors (activated Akt, survivin, transcription factor T cell factor 4 (TCF4), and its downstream target cyclin D1) and repression of proapoptotic factors (p53, active Bax, and cleaved caspase-3) in control mouse kidneys that were absent or aberrant in kidneys of Muc1 KO mice. Altogether, the data clearly indicate that Muc1 protection during acute kidney injury proceeds by enhancing both the HIF-1 and β-catenin protective pathways.
缺氧诱导因子(HIF)-1和β-连环蛋白保护途径是急性肾损伤时被激活的两种最重要的细胞反应。我们之前报道过,小鼠黏蛋白(Muc)1通过稳定HIF-1α、增强HIF-1下游信号传导,从而预防代谢应激,在缺血再灌注损伤(IRI)小鼠模型中保护肾功能和形态(Pastor-Soler等人,《Muc1在肾脏缺血再灌注损伤中具有保护作用》。《美国生理学杂志:肾脏生理学》308:F1452-F1462,2015年)。我们提出疑问,Muc1在IRI期间是否调节β-连环蛋白保护途径,原因如下:1)在培养的人类细胞中,β-连环蛋白的核定位依赖于MUC1;2)在培养细胞和组织的提取物中,β-连环蛋白与人类MUC1存在于共免疫沉淀复合物中;3)MUC1可防止糖原合酶激酶(GSK)3β介导的β-连环蛋白磷酸化,进而防止β-连环蛋白降解。使用相同的IRI小鼠模型,我们发现与Muc1基因敲除(KO)小鼠相比,对照小鼠肾脏中活性GSK3β的水平显著降低。因此,在恢复24小时和72小时时,对照小鼠肾脏中的β-连环蛋白显著上调,并在72小时时出现在核组分中。在Muc1 KO小鼠中,β-连环蛋白的诱导和核定位均不存在。我们还发现,对照小鼠肾脏中β-连环蛋白促生存因子(活化的Akt、生存素、转录因子T细胞因子4(TCF4)及其下游靶点细胞周期蛋白D1)的下游诱导以及促凋亡因子(p53、活化的Bax和裂解的半胱天冬酶-3)的抑制在Muc1 KO小鼠的肾脏中不存在或异常。总之,数据清楚地表明,急性肾损伤期间Muc1的保护作用是通过增强HIF-1和β-连环蛋白保护途径来实现的。