mTORC1 Regulation of Fibrogenesis, Amino Acid and Glucose Transport, Oxidative Stress, and Pro-inflammatory Genes
(A) Gene expression in kidney cortex extracts of 5-month-old RPTC-specific Tsc1-KO (Sglt2Cre;Tsc1fl/fl) mice compared with controls.
(B) 4E-BP1 phosphorylation.
(C and D) 4E-BP1 phosphorylation (C) and gene expression (D) in kidney cortex extracts of 5-month-old Akita mice and RPTC-specific Raptor-KO (Sglt2Cre;Raptorfl/+) Akita mice.
(E and F) Expression of collagen, amino acid transporters (E), and BCAA-degrading enzymes (F) in Akita mice treated with and without dapagliflozin for 3 months compared with wild-type (WT) controls.
(G) LLC-PK1 cells treated with 20 mM BCH or left untreated for 3 and 24 h. mTORC1 activity was analyzed using pS6 immunostaining. Quantification of fluorescence intensity is shown.
(H) Wild-type and Akita mice were intraperitoneally (i.p.) injected with BCH (2 μM/g body weight per day) for 5 days. mTORC1 activity was analyzed using immunostaining for pS6 and quantification of pS6 fluorescence intensity in RPTCs.
(I) A mechanistic model of the effects of diabetes and of treatment with SGLT2i on the development and progression of DKD (see text for details).
Scale bar, 50 μm. Data represent the mean ± SEM of three to five mice per group. ∗p < 0.05 and ∗∗p < 0.01 relative to the control wild-type or Akita groups; #p < 0.05 and ##p < 0.01 relative to the Akita control group; ππp < 0.01 relative to the BCH untreated control group.