Ursolic Acid Treatment Alleviates Diabetic Kidney Injury By Regulating The ARAP1/AT1R Signaling Pathway

Figures

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  Figure 1. General characteristics of db/db mice after UA treatment. (A) Weekly changes in the body weights of mice. (B, C) Blood glucose levels were measured in blood samples collected from the tail vein. (D, E) Changes in the blood glucose levels and urinary albumin/creatinine ratio in 20-week-old mice. ap<0.05 compared with db/m mice, bp<0.05 compared with diabetic db/db mice, and cp<0.01 compared with diabetic db/db mice. #p<0.05 compared with db/m mice, *p<0.05 compared with diabetic db/db mice, and **p<0.01 compared with diabetic db/db mice. db/m group, n=10; db/db group, n=10; and db/db+UA group, n=10.

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  Figure 2. UA treatment reduces renal histological changes in db/db mice. (A) Images of Masson’s trichrome staining of kidney tissues from 20-week-old mice (×400). (B) Semiquantitative analysis of the GSI as determined by Masson’s trichrome staining. (C) Images of PAS staining of kidney tissues from 20-week-old mice (×400). (D) Semiquantitative analysis of glomerular mesangial matrix hypertrophy as determined by PAS staining. (E) Electron microscopy images of basement membrane thickness (blue arrow) and the degree of podocyte foot process effacement (yellow arrow). #p<0.01 compared with db/m mice and *p<0.05 compared with diabetic db/db mice. db/m group, n=10; db/db group, n=10; and db/db+UA group, n=10.

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  Figure 3. Changes in ARAP1, AT1R, NOX2, 8-OHdG, FN, IL-1β and IL-18 expression in the kidney tissues of 20-week-old mice were assessed by immunohistochemistry. (A-G) Immunohistochemistry for ARAP1, AT1R, NOX2, 8-OHdG, FN, IL-1β and IL-18 expression. (H-N) Semiquantitative analysis of the immunohistochemistry results. #p<0.05, ####p<0.0001 compared with db/m mice and *p<0.05, **p<0.01 compared with diabetic db/db mice.

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  Figure 4. Protein and mRNA expression in the renal cortex of 20-week-old mice as detected by Western blotting and RT-qPCR, respectively. (A) Changes in ARAP1, AT1R, NOX2, NOX4, FN and Col IV mRNA expression as detected by RT-qPCR. (B) Changes in the levels of ARAP1, AT1R, NOX2, NOX4, FN, Col IV, and TGF-β1 proteins were detected using Western blotting. #p<0.01 compared with db/m mice; *p<0.05 and **p<0.01 compared with diabetic db/db mice.

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  Figure 5. UA treatment and ARAP1 knockdown reduced mesangial cell inflammation, extracellular matrix protein expression, fibrosis and oxidative stress. (A) Changes in ARAP1, AT1R, Col IV, IL-1β and IL-18 protein levels detected by Western blotting. (B–F) Semiquantitative analysis of Western blotting results. (G) Changes in the expression of FN and NOX2 in each group as assessed using fluorescence microscopy (x200). #P<0.05, ##P<0.01, ####P<0.0001 compared with the NG group, and *p<0.05 ****p<0.0001 compared with the HG group.