Xie Ying, Ruan Yuanyuan, Zou Huimei, Wang Yixin, Wu Xin, Li Xiaoying, Lai Jiao, Shi Mingjun, Xiao Ying, Wang Yuanyuan, Zhou Yuxia, Guo Bing, Zhang Fan
Department of Pathophysiology, Guizhou Medical University, Guiyang, China.
Key Laboratory of Pathogenesis and Drug Research of Common Chronic Diseases in Guizhou Province, Guizhou Medical University, Guiyang, China.
Pathobiology. 2021;88(6):412-423. doi: 10.1159/000517575. Epub 2021 Aug 3.
The goal of the present study was to determine the expression of yes-associated protein 1 (YAP1) in renal tissues of mice with lupus nephritis (LN) and elucidate its role in the progression of renal fibrosis.
C57BL/6 mice and MRL/lpr mice were selected for experimental comparison. Mouse kidney tissues were removed and sectioned for hematoxylin and eosin staining, Masson's trichome staining, Sirius staining, and immunohistochemistry. The mRNA and protein levels of YAP1 in mouse kidney tissues were detected, and the correlation between YAP1 and fibronectin (FN) mRNA levels was analyzed. Mouse renal epithelial cells were used for in vitro experiments. After transfection and stimulation, the cells were divided into 4 groups, namely the C57BL/6 serum group (group 1), the MRL/lpr serum group (group 2), the MRL/lpr serum + siRNA-negative control group (group 3), and the MRL/lpr serum + siRNA-YAP1 group (group 4). Epithelial-mesenchymal transition (EMT) markers in each group were detected by Western blotting and immunofluorescence staining. Serum creatinine, blood urea nitrogen, and urinary protein levels were detected and assessed for their correlation with YAP1 mRNA levels by Spearman's analysis.
Compared to C57BL/6 mice, MRL/lpr mice exhibited obvious changes in fibrosis in renal tissues. In addition, YAP1 expression was significantly higher in the renal tissues of MRL/lpr mice than in those of C57BL/6 mice, and YAP1 mRNA levels were positively correlated with those of FN. YAP1 silencing in lupus serum-stimulated cells could effectively relieve serum-induced EMT. Finally, we observed that YAP1 mRNA levels in mouse kidney tissue were significantly and positively correlated with the degree of renal function injury.
YAP1 expression in the kidney tissues of LN mice was higher than that observed in normal mice, indicating that YAP1 may play an important role in the occurrence and development of LN.
本研究旨在确定Yes相关蛋白1（YAP1）在狼疮性肾炎（LN）小鼠肾组织中的表达，并阐明其在肾纤维化进展中的作用。
选取C57BL/6小鼠和MRL/lpr小鼠进行实验对比。取小鼠肾脏组织，进行切片，用于苏木精-伊红染色、Masson三色染色、天狼星染色和免疫组织化学。检测小鼠肾组织中YAP1的mRNA和蛋白水平，并分析YAP1与纤连蛋白（FN）mRNA水平之间的相关性。采用小鼠肾上皮细胞进行体外实验。转染和刺激后，将细胞分为4组，即C57BL/6血清组（第1组）、MRL/lpr血清组（第2组）、MRL/lpr血清+siRNA阴性对照组（第3组）和MRL/lpr血清+siRNA-YAP1组（第4组）。通过蛋白质印迹法和免疫荧光染色检测各组上皮-间质转化（EMT）标志物。检测血清肌酐、血尿素氮和尿蛋白水平，并通过Spearman分析评估其与YAP1 mRNA水平的相关性。
与C57BL/6小鼠相比，MRL/lpr小鼠肾组织纤维化有明显变化。此外，MRL/lpr小鼠肾组织中YAP1表达明显高于C57BL/6小鼠，且YAP1 mRNA水平与FN mRNA水平呈正相关。狼疮血清刺激细胞中YAP1沉默可有效缓解血清诱导的EMT。最后，我们观察到小鼠肾组织中YAP1 mRNA水平与肾功能损伤程度呈显著正相关。
LN小鼠肾组织中YAP1表达高于正常小鼠，表明YAP1可能在LN的发生发展中起重要作用。
