Macrophage Depletion Lowered Blood Pressure and Attenuated Hypertensive Renal Injury and Fibrosis

Figures

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  FIGURE 1. Treatment with liposome-encapsulated clodronate (LEC) reduced systolic blood pressure (SBP) in the angiotensin (Ang) II mice (A) and DS rats (B) and attenuated the ratio of albuminuria/creatinine (C) in the Ang II mice. Quantification of monocyte, lymphocyte, and granulocyte population in the circulating peripheral blood smear stained with Giemsa in Ang II mice (D) and hypertensive DS rats (E). Ctr, control mice treated with PBS liposome; LEC, normal mice treated with LEC; Ang II, the mice treated with Ang II (1400 ng/min/mg body weight); Ang/LEC, the mice treated with Ang II plus LEC treatment; NS, DS rats fed a normal salt (0.5% NaCl) diet; HS, DS rats fed a HS (4% NaCl) diet. N = 8, ∗p < 0.05, vs. control group or NS group, #p < 0.05, vs. Ang II group or HS group.

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  FIGURE 2. LEC treatment effectively reduced renal macrophages in Ang II mice. (A) Representative images of MOMA2 expression evaluated by immunofluorescence staining. (B) Quantitative assessment of MOMA2 expression in each group of mice. Green color indicates MOMA2 and blue color indicates DAPI. N = 8, ∗p < 0.05, vs. control group, #p < 0.05 vs. Ang II group.

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  FIGURE 3. The macrophage depletion by liposome-encapsulated clodronate (LEC) attenuated Ang II-induced injury. (A) Representative images of kidney section stained with periodic acid-Schiff (PAS) for evaluation of glomerular injury. (B) Quantitative assessment of glomerular sclerosis in each group of mice. N = 8, ∗p < 0.05, vs. control group, #p < 0.05, vs. Ang II group.

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  FIGURE 4. The macrophage depletion by LEC attenuated Ang II-induced renal fibrosis. (A) Representative the images of Masson-Trichrome-stained kidney sections for evaluation of renal fibrosis. (B) Quantitative assessment of renal fibrosis in each group of mice. N = 8, ∗p < 0.05, vs. control group, #p < 0.05, vs. Ang II group.

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  FIGURE 5. The macrophage depletion by LEC protected against renal injury and fibrosis in hypertensive DS rats. (A) Representative images of kidney section stained with periodic acid-Schiff (PAS) for evaluation of glomerular injury. (B) Quantitative assessment of glomerular sclerosis in each group of mice. (C) Representative images of kidney section stained with PAS for evaluation of glomerular injury. (D) Quantitative assessment of glomerular sclerosis in each group of mice. NS, DS rats were fed a normal salt diet; HS, DS rats were fed a HS diet; HS/LEC, DS rats were fed a HS diet with LEC treatment. N = 8, ∗p < 0.05, vs. NS group, #p < 0.05, vs. HS group.

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  FIGURE 6. The macrophage depletion by LEC attenuated the protein expression of proinflammatory cytokines tumor necrosis factor (TNF) α (A) and interleukin β1 (B) in the kidney of the Ang II mice. N = 8, ∗p < 0.05, vs. control group, #p < 0.05, vs. Ang II group.

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  FIGURE 7. Macrophage depletion by LEC attenuated the protein expression of profibrotic factors tissue growth factor (TGF) β1 (A) and fibronectin (B) in the kidney of the Ang II mice. N = 8, ∗p < 0.05, vs. control group, #p < 0.05, vs. Ang II group.

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  FIGURE 8. The macrophage depletion by LEC reduced oxidative fluorescence densities in the Ang II mice. (A) Representative images of renal sections stained with DHE for the evaluation of oxidative fluorescence density evaluated by confocal fluorescence microscope. (B) Quantification of average fluorescence densities in each group of mice. N = 8, ∗p < 0.05, vs. control group, #p < 0.05, vs. Ang II group.

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  FIGURE 9. Effects of LEC treatment on NADPH oxidative activity (A), the protein expression of renal NADPH oxidase subunits gp91phox (B) and p22phox (C), and phospho-eNOS (Ser 1177, D) in Ang II mice. The macrophage depletion by LEC reduced renal NADPH oxidase activity and the expression of NADPH subunits gp91phox and p22phox, and preserved phospho-eNOS (Ser 1177) in the Ang II mice. N = 8, ∗p < 0.05, vs. control group, #p < 0.05, vs. Ang II group.