Deletion of Akt1 Promotes Kidney Fibrosis in a Murine Model of Unilateral Ureteral Obstruction

Figure 3

Akt1 deletion is associated with upregulation of TGFβ1. (a) Immunohistochemical staining (×200). Expression of TGFβ1 began to increase from day 1 after UUO in Akt1^−/− mice. TGFβ1 was expressed more strongly in Akt1^−/− mice than in wild type mice as UUO progressed. (b) Western blot analysis. TGFβ1 expression was higher in Akt1^−/− mice than in wild type mice on day 0 (U0 vs. A0, ) and day 1 (U1 vs. A1, ) but was not statistically different between the two groups on day 3 (U3 vs. A3, ) and day 7 (U7 vs. A7, ). (c) Western blot analysis. In HK2 cells, silencing of Akt1 promoted the expression of TGFβ1 and fibronectin in the unstimulated state, which was augmented by angiotensin II stimulation (1 μM). (d) Western blot analysis. In NRK-49F cells, silencing of Akt1 did not affect the expression of TGFβ1 and fibronectin regardless of angiotensin II stimulation. Abbreviations: ANGII: angiotensin II; GAPDH: glyceraldehyde 3-phosphate dehydrogenase; HK2 cells: immortalized human proximal tubular cells; NRK-49F cells: rat kidney fibroblasts; TGFβ1: transforming growth factor β1; UUO: unilateral ureteral obstruction; A0 (): Akt1^−/− sham; A1 (): day 1 Akt1^−/−; A3 (): day 3 Akt1^−/−; A7 (): day 7 Akt1^−/−; U0 (): wild type sham; U1 (): day 1 wild type; U3 (): day 3 wild type; U7 (): day 7 wild type.

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