基于“肠-肾轴”探索Pellino1促进肾小管上皮细胞胞外转运对甲酚葡糖苷酸(p-CG)抑制脓毒症急性肾损伤发生发展的作用及机制

肠道微生物菌群稳态近年来被发现与急性肾损伤（AKI）密切相关，其中肾小管上皮细胞内尿毒症毒素对甲酚葡糖苷酸（p-CG）的沉积是导致肾小管不可逆损伤的重要因素。我们前期研究发现Pellino1在脓毒症AKI小鼠肾小管上皮细胞中表达下调并与p-CG胞外转运受体ABCC4和ABCG2正相关。Pellino1敲除能显著抑制ABCC4和ABCG2基因转录及启动子招募CREBBP、ARNTL、PRDM4、NPAS2和CEBPα等5个目标转录因子（TF）。本项目拟通过DLR、EMSA、Co-IP、GST-pulldown、质谱-泛素化和CRISPR-Cas9基因编辑等技术剖析Pellino1调控该5个目标TF促进ABCC4和ABCG2转录的分子机制及各目标TF在Pellino1抑制脓毒症AKI发展中的作用，结果有望阐明Pellino1调控p-CG胞外转运的机制并为脓毒症AKI治疗提供新靶点，意义重大。

Gut microbiota homeostasis has been found to be tightly associated with acute kidney injury (AKI) in recent years, and deposition of p-cresyl-glucuronide (p-CG) in tubular epithelial cells was identified as an important factor for the irreversible tubular damage. Our previous study unveiled that Pellino1 was downregulated in tubular epithelial cells of mice with septic AKI and such downregulation was positively correlated with two efflux transporters of p-CG, ABCC4 and ABCG2. Pellino1 knockout was able to repress transcription of ABCC4 and ABCG2 as well as their promoters to recruit five target transcription factor (TF) including CREBBP, ARNTL, PRDM4, NPAS2 and CEBPα. The current project aims at depicting the molecular mechanisms of how Pellino1 regulate the five target TF to promote transcription of ABCC4 and ABCG2 and clarifying their influences in the anti-septic AKI roles of Pellino1 through DLR, EMSA, Co-IP, GST-pulldown, Mass spectrometry-ubiquitination and CRISPR-Cas9 gene editing technologies. Our project is in hope of demonstrating the mechanism whereby Pellino1 facilitates efflux transportation of p-CG in tubular epithelial cells and offering novel target for septic AKI management.