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Abstract: PO0697

Metabolic Images Using Fluorescence Lifetime Imaging Reveals Metabolic Alteration in Proximal Tubular Epithelial Cells in Type 2 Diabetes

Session Information

Category: Diabetic Kidney Disease

  • 601 Diabetic Kidney Disease: Basic

Authors

  • Kwon, Woo Young, Division of Nephrology, Department of Internal Medicine, Kyung Hee University, College of Medicine, Seoul, Korea (the Republic of)
  • Jung, Gun Tae, Department of Biomedical Science and Technology, Kyung Hee Medical Science Research Institute, Kyung Hee University, Seoul, Korea (the Republic of)
  • Jung, Su Woong, Division of Nephrology, Department of Internal Medicine, Kyung Hee University, College of Medicine, Seoul, Korea (the Republic of)
  • Kim, Yang gyun, Division of Nephrology, Department of Internal Medicine, Kyung Hee University, College of Medicine, Seoul, Korea (the Republic of)
  • Lee, Sangho, Division of Nephrology, Department of Internal Medicine, Kyung Hee University, College of Medicine, Seoul, Korea (the Republic of)
  • Kim, Kwang Pyo, Department of Applied Chemistry, Institute of Natural Science, Global Center for Pharmaceutical Ingredient Materials, Kyung Hee University, Yongin, Korea (the Republic of)
  • Chae, Weon-Sik, Daegu Center, Korea Basic Science Institute, Daegu, Korea (the Republic of)
  • Moon, Ju young, Division of Nephrology, Department of Internal Medicine, Kyung Hee University, College of Medicine, Seoul, Korea (the Republic of)
Background

Although there is a massive metabolic alteration of the kidney in diabetes, it is difficult to detect and measure the single-cell nicotinamide adenine dinucleotide hydrogen (NADH), flavin adenine dinucleotide (FAD) production, and redox potential. In particular, proximal tubular epithelial cells (PTECs) are the most laborious and affected cells under high glucose environments. We investigated this study to evaluate quantitative PTECs-specific metabolic images in the diabetic kidney using fluorescence lifetime imaging (FLIM).

Methods

Kidney sections of 20 week-old db/db and db/m mice were used for FLIM. FLIM images are analyzed using the phasor approach. The FLIM image and phasor plot representing FLIM data in vector space were measured through Leica TCS SP8 SMD and LAS-X software. The NADH, FAD, and ATP levels in diabetic kidneys were measured using LC-MS analysis by Q-trap 5500.

Results

NADH and FAD located at the different subcellular levels in PTECs. The NADH phasor analysis of PTECs revealed a right-ward shift toward shorter lifetimes from the db/m to the db/db, while there was no significant alteration of FAD between the two groups. It could be indicative of an increase in the NADH-to-FAD ratio that alters metabolic flux. In addition, the levels of NADH in diabetic kidneys were significantly increased than db/m, while the levels of FAD were reduced in diabetic kidneys. Finally, ATP level decreased in the diabetic kidney compared to db/m.

Conclusion

NADH and FAD FLIM in PTECs is an optimal approach to characterize and monitor metabolism in diabetic kidneys. Quantitative metabolic imaging using FLIM enables to measure and analyze metabolic alteration with spatial information.