鈉-葡萄糖協同轉運蛋白2抑制劑對腎結石的抑制效果：從細胞及動物實驗探討分子機轉

劉展榮、何冠達、黃鶴翔、蔡曜聲¹

國立成功大學附設醫院 泌尿部¹

The Sodium-Glucose Cotransporter-2 Inhibitor Dapagliflozin Inhibit Kidney Stone Formation Through Modulating AMPK-Mediated Autophagy

Chan-Jung Liu, Kuan-Ta Ho², Ho-Shiang Huang¹, Yau-Sheng Tsai^2,3

Department of Urology, National Cheng Kung University Hospital, Tainan, Taiwan¹; Institute of Clinical Medicine, College of Medicine, National Cheng Kung University, Taiwan²; Center for Clinical Medicine Research, National Cheng Kung University Hospital, Tainan, Taiwan³

Purpose: Diabetes mellitus (DM) and metabolic syndrome are all associated with increased risk of urolithiasis. Despite the advance in surgical technique for urolithiasis, effective medical therapies remain lacking. Sodium-glucose cotransporter 2 inhibitor (SGLT2i), a novel antidiabetic drug, can not only control hyperglycemia via increasing glycosuria but also reduce oxidative stress with cardiorenal benefits independent of glucose control. Due to the antioxidative and diuretic effects, SGLT2i could potentially reduce the risk of urolithiasis. Although a recent cohort study found antilithic association with SGLT2i, the exact mechanism remains unclear. In the present study, we sought to investigate the effects of dapagliflozin (DAPA) on the kidney stone formation and study the mechanisms of DAPA-provided antilithic ability.

Materials and Methods: For in intro studies, human renal tubular (HK-2) cells were exposed to oxalate (0.2 mM) and calcium oxalate monohydrate (COM) (10 ug/cm²) for 12 hours, treated with and without DAPA (20uM). For in vivo investigations, we used glyoxylate (GOX) intraperitoneal injection on male mice for 7 days to establish hyperoxaluric urolithiasis model. Oral gavage with 10mg/kg/d DAPA for 7 days in the meantime.

Results: Results from in vitro experiments showed that ROS and apoptosis, as well as NLRP3 inflammasome pathway (phosphorylated NFkB, ASC, pro CASPASE 1) and autophagy activity (LC3B 1 conversion to LC3B2, AMPKα, p62), are significantly increased after co-exposure to Ox and COM. Furthermore, the expression of phosphorylated AMPK elevated in HK-2 cells following DAPA treatment under Ox + COM co-exposure. treatment of DAPA significantly suppressed ROS generation (demonstrated by CM-H2DCFDA staining), NLRP3 inflammasome activation (decreased phosphorylated NFkB), and autophagy activity (decreased p62 and LC3B2). Meanwhile, results from in vivo study also demonstrated the expression of AMPK, BECN1, LC3B1, and p62 all increased in GOX group. However, DAPA treatment diminished the expression of AMPK but increased phosphorylated AMPK. Furthermore, we demonstrated that DAPA decreased CaOx crystals deposits in kidney through polarized microscopy.

Conclusions: our results suggested that DAPA treatment holds the potential to decrease CaOx stone formation via restoring phosphorylated AMPK expression and modulating kidney autophagy activity.