Both Specific Endothelial and Proximal Tubular Adam17 Deletion Protect against Diabetic Nephropathy

Figure 1. Glomerular tuft area, mesangial area, mesangial index and podocyte loss. (A) Glomerular tuft area from eAdam17 model. (B) Mesangial expansion from eAdam17 model. (C) Mesangial index from eAdam17 model. (D) Glomerular tuft area from tAdam17 model. (E) Mesangial expansion from tAdam17 model. (F) Mesangial index from tAdam17 model. (G) Representative images of the PAS staining for eAdam17 and tAdam17 mice. Original magnification, ×400. (H) Podocyte number is represented as the number of brown positive cells after WT-1 immunostaining in eAdam17 model. (I) Podocyte number is represented as the number of brown positive cells after WT-1 immunostaining in tAdam17 model. (J) Representative images of the WT-1 immunostaining for eAdam17 and tAdam17 mice. Original magnification, ×400. CONT, control; DB, diabetic; eAdam17, endothelial ADAM17; KO, knockout; tAdam17, proximal tubular ADAM17; WT, wild-type; WT-1, wilms tumor 1. * p < 0.05 DB vs. NoDB, $ p < 0.05 KO vs. WT; n = 6–9 in each group.

Figure 2. Effect of diabetes and endothelial or proximal-tubular Adam17 deletion on the Renin Angiotensin System. (A) Circulating ACE2 activity from eAdam17 model. (B) Circulating ACE2 activity from tAdam17 model. (C) Renal ACE2 activity from eAdam17 model. (D) Renal ACE2 activity from tAdam17 model. (E) At1r gene expression from eAdam17 model. (F) At1r gene expression from tAdam17 model. (G) At2r gene expression from eAdam17 model. (H) At2r gene expression from tAdam17 model. ACE2, angiotensin converting enzyme 2; AT1R, angiotensin II receptor type 1; AT2R, angiotensin II receptor type 2; CONT, control; DB, diabetic; eAdam17, endothelial ADAM17; KO, knockout; tAdam17, proximal tubular ADAM17; WT, wild-type. * p < 0.05 DB vs. NoDB, $ p < 0.05 KO vs. WT; n = 6–8 in each group.

Figure 3. Influence of diabetes and Adam17 deletion on circulating TNF-α, Tnfa, Tnfr1, and Ccl5 gene expression. (A) Circulating TNF-α from eAdam17 model. (B) Circulating TNF-α from tAdam17 model. (C) Tnfa gene expression from eAdam17 model. (D) Tnfa gene expression from tAdam17 model. (E) Tnfr1 gene expression from eAdam17 model. (F) Tnfr1 gene expression from tAdam17 model. (G) Ccl5 gene expression from eAdam17 model. (H) Ccl5 gene expression from tAdam17 model. CONT, control; DB, diabetic; eAdam17, endothelial ADAM17; KO, knockout; tAdam17, proximal tubular ADAM17; TNF-α, tumor necrosis factor α; WT, wild-type. * p < 0.05 DB vs. NoDB, $ p < 0.05 KO vs. WT; n = 6–8 in each group.

Figure 4. Influence of diabetes and Adam17 deletion on F4/80 immunostaining. (A) F4/80 immunostaining from eAdam17 model. (B) F4/80 immunostaining from tAdam17 model. Original magnification, ×200. CONT, control; DB, diabetic; eAdam17, endothelial ADAM17; KO, knockout; tAdam17, proximal tubular ADAM17; TNF-α, tumor necrosis factor α; WT, wild-type. * p < 0.05 DB vs. NoDB, $ p < 0.05 KO vs. WT; n = 6–8 in each group.

Figure 5. Influence of diabetes and Adam17 deletion on Tgfα, Tgfβ, and fibronectin gene expression and protein expression. (A) Tgfα gene expression from the eAdam17 model. (B) Tgfα gene expression from the tAdam17 model. (C) Tgfβ gene expression from the eAdam17 model. (D) Tgfβ gene expression from the tAdam17 model. (E) TGF-β protein expression from eAdam17 model. (F) TGF-β protein expression from tAdam17 model. (G) Fibronectin gene expression from the eAdam17 model. (H) Fibronectin gene expression from the tAdam17 model. CONT, control; DB, diabetic; eAdam17, endothelial ADAM17; FN, fibronectin; KO, knockout; tAdam17, proximal tubular ADAM17; TGF-α, transforming growth factor α, TGF-β, transforming growth factor β; WT, wild-type. * p < 0.05 DB vs. NoDB, $ p < 0.05 KO vs. WT; n = 6–8 in each group.

Figure 6. Influence of diabetes and Adam17 deletion on α-SMA and collagen staining and pAkt/Akt ratio. (A) α-SMA immunostaining from eAdam17 model. (B) α-SMA immunostaining from tAdam17 model. (C) Representative images of α-SMA immunostaining from eAdam17 and tAdam17 models. Original magnification, ×100. (D) Collagen staining from eAdam17 model. (E) Collagen staining from tAdam17 model. (F) Representative images of Sirius Red staining from eAdam17 and tAdam17 models. Original magnification, ×100. (G) pAkt/Akt ratio from eAdam17 model. (H) pAkt/Akt ratio from tAdam17 model. CONT, control; DB, diabetic; eAdam17, endothelial ADAM17; KO, knockout; pAkt, phosphorylated protein kinase B; α-SMA, alpha smooth muscle actin; tAdam17, proximal tubular ADAM17; WT, wild-type. * p < 0.05 DB vs. NoDB, $ p < 0.05 KO vs. WT; n = 6–8 in each group.

Figure 7. Influence of high glucose medium and Adam17 deletion on α-SMA and Collagen-IV immunofluorescence staining. (A) α-SMA immunofluorescence staining in HKC-8 spheroids. (B) Representative images of the α-SMA immunofluorescence staining. (C) Type IV collagen immunofluorescence staining in HKC-8 spheroids. (D) Representative images of the Type IV collagen immunofluorescence staining. LG, low glucose; HG, high glucose; M, mannitol. * p < 0.05 HG vs. LG, $ p < 0.05 KO vs. WT, # p < 0.05 M vs. HG; n = 15–21 spheroids per group. Scale bar 20 µm.

Figure 8. Schematic figure summarizing the findings regarding the effects of Adam17 deletion in endothelial and proximal tubular cells under STZ-diabetic mice model.