THE EFFECT OF GENISTEIN TO REDUCE ALBUMINURIA DUE TO ALBUMIN ENDOCYTOSIS THROUGH VACEOLAE INTO GLOMERULAR ENDOTHELIAL AND EPITHELIAL CELLS BY DECREASING SRC KINASE ACTIVITY

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https://storage.unitedwebnetwork.com/files/1099/f1b8809704632c2d66876565331eaf78.pdf

Abstract Title *

THE EFFECT OF GENISTEIN TO REDUCE ALBUMINURIA DUE TO ALBUMIN ENDOCYTOSIS THROUGH VACEOLAE INTO GLOMERULAR ENDOTHELIAL AND EPITHELIAL CELLS BY DECREASING SRC KINASE ACTIVITY

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Co-author 1

Takahito Moriyama moriyama.takahito.7j@tokyo-med.ac.jp Tokyo Medical University Nephrology Tokyo Japan *

Co-author 2

Suguru Saito ss1104ss@tokyo-med.ac.jp Tokyo Medical University Nephrology Tokyo Japan -

Co-author 3

Yuta Kimura yk615615@tokyo-med.ac.jp Tokyo Medical University Nephrology Tokyo Japan -

Co-author 4

Sho Rinno rinno1216@gmail.com Tokyo Medical University Nephrology Tokyo Japan -

Co-author 5

Rieko China rieko6_6@tokyo-med.ac.jp Tokyo Medical University NephrologyNephrology Tokyo Japan -

Co-author 6

Yoshitaka Miyaoka mitaku38@tokyo-med.ac.jp Tokyo Medical University Nephrology Tokyo Japan -

Co-author 7

Miho Nagai m-nagai@tokyo-med.ac.jp Tokyo Medical University Nephrology Tokyo Japan -

Co-author 8

Yoshihiko Kanno kannoyh@tokyo-med.ac.jp Tokyo Medical University Nephrology Tokyo Japan -

Co-author 9

Co-author 10

Co-author 11

Co-author 12

Co-author 13

Co-author 14

Co-author 15

Introduction

Previously, we have been reported about the new etiology of albuminuria by caveolar mediated intracellular trafficking pathway of albumin in glomerular endothelial cells (GEnCs) and podocytes (Pod), and also researched novel therapeutic approach to reduce albuminuria by decreasing caveolae. Src kinase plays an important role of internalization of caveolae coated pits into cytoplasm as caveolae endocytosis. In this study, we analyzed whether Genistein (Gen), which was the non-steroidal phytoestrogen belonged to potent soy isoflavone, inhibited Src activity in GEnCs and pod, and resulted in decreasing albuminuria of puromycin aminonucleoside (PAN) induced nephrotic syndrome modeled mice.

Methods

In vitro study, GEnCs or Pod were treated with 50, 100, or 200 µM of Gen for 60min, then each cell was co-incubated with albumin for 15, 30, and 60 min. After that, the expression of albumin, caveolin-1 (Cav-1), and p-Src were evaluated by immunofluorescence (IF) and western blots (WB) analysis. In vivo study, PAN mice were treated by Gen or DMSO (as vehicle control) for 7 days. Then, the amount of albuminuria at day 7 between both groups and between pre and post treatments in each group were compared, and p-Src and Cav-1 expressions in glomeruli by IF and immunohistochemistry (IHC) were also compared among normal control, vehicle control, and Gen treated group.

Results

In each cell, albumin, Cav-1, and p-Src expression levels were significantly decreased by dose dependent manner of Gen in IF and WB analysis. In the in vivo study, albuminuria in vehicle control was significantly increased from day 0 to 7 (p=0.021), but it was not increased by Gen treatment. Moreover, albuminuria at day 7 was significantly lower by Gen than vehicle control (p=0.021). The p-Src and Cav-1 expressions in glomeruli were significantly increased in vehicle control (p<0.001), but not increased in Gen treated group rather than cormal control.

Conclusion

Gen interfered with albumin endocytosis through caveolae by suppressing caveolae expression according to inhibit Src kinase and resulted in reduced albuminuria in PAN mice. These results indicated the possibility of Src kinase inhibitor, Gen as the new therapeutic approach to reduce albuminuria in Nephrotic syndrome. This study was already presented in ASN Kidney Week 2026 at Houston, USA.

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