Priyamvada Shubha, Jayawardena Dulari, Anbazhagan Arivarasu N, Kumar Anoop, Saksena Seema, Gill Ravinder K, Borthakur Alip, Alrefai Waddah A, Dudeja Pradeep K
Division of Gastroenterology & Hepatology, Department of Medicine, University of Illinois Chicago, Chicago, Illinois, USA.
Jesse Brown VA Medical Center, Chicago, Illinois, USA.
J Cell Mol Med. 2025 May;29(9):e70513. doi: 10.1111/jcmm.70513.
Putative anion transporter 1 (PAT-1) is the key oxalate-secreting transporter in the intestine and therefore, maintaining its steady-state levels is critical for oxalate homeostasis. Autophagy is known to modulate the expression of intestinal solute transporters; however, its role in regulating PAT-1 has not been examined. Autophagy in Caco-2 cells was induced by either rapamycin treatment or by nutrient deprivation and assessed by conventional autophagy marker proteins. ATG7 (autophagy-related 7) protein expression was attenuated by ATG7-siRNA in Caco-2 cells or by utilising ATG7KO mice. PAT-1 protein levels in Caco-2 cells were significantly reduced by rapamycin or by nutrient deprivation at 48 and 72 h. Concomitantly, the LC3II/I ratio was increased, and p62 levels were significantly decreased, confirming the induction of autophagy. Nutrient deprivation for 6 h also caused a significant decrease in the surface levels of PAT-1. PAT-1 protein levels were increased by the siRNA-mediated ATG7 knockdown in Caco-2 cells and in the ileum of ATG7KO mice. In summary, Autophagy in intestinal epithelial cells modulates the basal levels of PAT-1 protein and may play a critical role in the maintenance of oxalate homeostasis.
推定阴离子转运蛋白1(PAT-1)是肠道中关键的草酸盐分泌转运蛋白,因此,维持其稳态水平对于草酸盐稳态至关重要。已知自噬可调节肠道溶质转运蛋白的表达;然而,其在调节PAT-1中的作用尚未得到研究。通过雷帕霉素处理或营养剥夺诱导Caco-2细胞中的自噬,并通过传统的自噬标记蛋白进行评估。在Caco-2细胞中,通过ATG7-siRNA或利用ATG7基因敲除小鼠使ATG7(自噬相关7)蛋白表达减弱。在48小时和72小时时,雷帕霉素或营养剥夺显著降低了Caco-2细胞中PAT-1蛋白水平。同时,LC3II/I比率增加,p62水平显著降低,证实了自噬的诱导。6小时的营养剥夺也导致PAT-1表面水平显著降低。在Caco-2细胞和ATG7基因敲除小鼠的回肠中,siRNA介导的ATG7敲低使PAT-1蛋白水平升高。总之,肠道上皮细胞中的自噬调节PAT-1蛋白的基础水平,并可能在维持草酸盐稳态中起关键作用。