FOXO3a Protects against Kidney Injury in Type II Diabetic Nephropathy by Promoting Sirt6 Expression and Inhibiting Smad3 Acetylation

Figure 5

Downregulation of FOXO3a leads to the deterioration of kidney injury in db/db mice. (a, b) Protein and mRNA were extracted from the kidneys of 20-week-old db/db and db/+ mice. (a) FOXO3a mRNA expression analyzed by qPCR. (b) FOXO3a protein expression assessed by western blot analysis. (c–j) The db/db or db/+ mice used in all experiments were 20 weeks old and were infected with sh-NC or sh-FOXO3a lentiviruses in the kidneys at 18 weeks old to knock down FOXO3a expression in vivo. (c–e) Graphs showing ACR, BUN, and proteinuria as analyzed by the ELISA kit. (f) Representative images of PAS staining of kidneys from mice. Scale bar, 40 μm. (g) Scoring of tubular injury. Proximal and distal tubules are marked with red and black arrows, respectively. “^” represents the brush border of proximal tubules. “” represents the cellular debris in the lumen of proximal tubules. “” represents the disorder proximal tubular structure, respectively. (h) IHC analysis of α-SMA, E-cadherin, and FOXO3a expression in kidney tissue. (i) α-SMA, E-cadherin, FOXO3a, and Collagen I mRNA expression analyzed by qPCR. (j) Protein expression of α-SMA, E-cadherin, FOXO3a, and Collagen I assessed by western blot analysis. (k) KIM-1 and NGAL mRNA expression analyzed by qPCR. (l) Protein expression of KIM-1 and NGAL assessed by western blot analysis. The data are presented as the . experiments in (a–e) and (h–l). experiments in (g). , , and .

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