Renal Section, Department of Medicine, Boston University School of Medicine, Boston, Massachusetts, USA; Whitaker Cardiovascular Institute, Boston University, Boston, Massachusetts, USA.
Renal Section, Department of Medicine, Boston University School of Medicine, Boston, Massachusetts, USA.
Kidney Int. 2020 Mar;97(3):538-550. doi: 10.1016/j.kint.2019.09.029. Epub 2019 Oct 30.
Emerging evidence in animal models of chronic kidney disease (CKD) implicates Aryl Hydrocarbon Receptor (AHR) signaling as a mediator of uremic toxicity. However, details about its tissue-specific and time-dependent activation in response to various renal pathologies remain poorly defined. Here, a comprehensive analysis of AHR induction was conducted in response to discrete models of kidney diseases using a transgenic mouse line expressing the AHR responsive-promoter tethered to a β-galactosidase reporter gene. Following validation using a canonical AHR ligand (a dioxin derivative), the transgenic mice were subjected to adenine-induced and ischemia/reperfusion-induced injury models representing CKD and acute kidney injury (AKI), respectively, in humans. Indoxyl sulfate was artificially increased in mice through the drinking water and by inhibiting its excretion into the urine. Adenine-fed mice showed a distinct and significant increase in β-galactosidase in the proximal and distal renal tubules, cardiac myocytes, hepatocytes, and microvasculature in the cerebral cortex. The pattern of β-galactosidase increase coincided with the changes in serum indoxyl sulfate levels. Machine-learning-based image quantification revealed positive correlations between indoxyl sulfate levels and β-galactosidase expression in various tissues. This pattern of β-galactosidase expression was recapitulated in the indoxyl sulfate-specific model. The ischemia/reperfusion injury model showed increase in β-galactosidase in renal tubules that persisted despite reduction in serum indoxyl sulfate and blood urea nitrogen levels. Thus, our results demonstrate a relationship between AHR activation in various tissues of mice with CKD or AKI and the levels of indoxyl sulfate. This study demonstrates the use of a reporter gene mouse to probe tissue-specific manifestations of uremia in translationally relevant animal models and provide hypothesis-generating insights into the mechanism of uremic toxicity that warrant further investigation.
在慢性肾脏病 (CKD) 的动物模型中出现的新证据表明,芳香烃受体 (AHR) 信号作为尿毒症毒性的介质。然而,其对各种肾脏病变的组织特异性和时间依赖性激活的细节仍未得到明确界定。在这里,我们使用表达与 β-半乳糖苷酶报告基因相连的 AHR 反应启动子的转基因小鼠系,对针对不同肾脏疾病的特定模型进行了 AHR 诱导的全面分析。在用经典的 AHR 配体(二恶英衍生物)进行验证后,将转基因小鼠分别置于腺嘌呤诱导的和缺血/再灌注诱导的损伤模型中,分别代表人类的 CKD 和急性肾损伤 (AKI)。通过饮用水和抑制其向尿液中的排泄,人为地增加了小鼠体内的硫酸吲哚酚。腺嘌呤喂养的小鼠在近端和远端肾小管、心肌细胞、肝细胞和大脑皮质微血管中,β-半乳糖苷酶的表达明显增加。β-半乳糖苷酶增加的模式与血清硫酸吲哚酚水平的变化一致。基于机器学习的图像定量显示,血清硫酸吲哚酚水平与各种组织中的β-半乳糖苷酶表达之间存在正相关。这种β-半乳糖苷酶表达模式在硫酸吲哚酚特异性模型中得到了重现。缺血/再灌注损伤模型显示,尽管血清硫酸吲哚酚和血尿素氮水平降低,但肾小管中的β-半乳糖苷酶仍增加。因此,我们的结果表明,在 CKD 或 AKI 小鼠的各种组织中,AHR 的激活与硫酸吲哚酚的水平之间存在关联。本研究证明了使用报告基因小鼠在转化相关动物模型中探测尿毒症的组织特异性表现,并为尿毒症毒性的机制提供了假说生成的见解,值得进一步研究。