Preclinical Investigations on Anti-fibrotic Potential of Long-Term Oral Therapy of Sodium Astragalosidate in Animal Models of Cardiac and Renal Fibrosis

Figures

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  Figure 1. Effect of SA on ISO-induced myocardial injury and fibrosis rats. (A) Outline of experimental protocol to evaluate the effect of SA against ISO-induced cardiac fibrosis (n = 8). (B) H&E and Masson staining shows the cardiac structural damage of myocardial fibrosis induced by ISO in rats (100× and 400× magnification). (C) Collagen volume fraction quantified from the stained sections. (D–F) Effect of SA on serum levels of IL-6, TNF-α, and IL-1β of myocardial fibrosis in rats. All data are represented as the mean ± SEM. *p < 0.05, **p < 0.01, and ***p < 0.001 compared with the control group; #p < 0.05, ##p < 0.01, and ###p < 0.001 compared with the ISO group.

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  Figure 2. Effect of SA on ISO-induced fibrotic gene expression in heart tissues. (A) qRT-PCR analysis of α-SMA and collagen I/III mRNA expression in ISO-induced myocardial fibrosis in rats. (B–G) qRT-PCR analysis of MMP-2/9 and TIMP-1/2 mRNA expression in ISO-induced myocardial fibrosis in rats. All data are represented as the mean ± SEM. *p < 0.05, **p < 0.01, and ***p < 0.001 compared with the control group; #p < 0.05, ##p < 0.01, and ###p < 0.001 compared with the ISO group.

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  Figure 3. Oral administration of SA inhibited lipid metabolism impairment in db/db mice. (A) Outline of experimental protocol to evaluate the effect of SA against renal fibrosis in db/db mice (n = 8). (B, C) Effects of SA on body weight and blood glucose in db/db mice (n = 8). (D–G) The levels of TC, TG, LDL, and HDL in serum were analyzed in the experiment (n = 8). (H) Illustration of H&E staining of the liver tissue in different groups. Magnification 400×. All data are represented as the mean ± SEM. *p < 0.05, **p < 0.01, and ***p < 0.001 compared with the db/m group; #p < 0.05, ##p < 0.01, and ###p < 0.001 compared with the db/db mice.

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  Figure 4. Effects of SA on kidney structure and function in db/db mice. (A) Representative pictures of the right kidneys from each group. (B) Ratio of kidney weight/body weight. (C–E) Serum levels of BUN, MAU and ACR in db/db mice (n = 8). (F) H&E staining (100× magnification) and Masson staining (400× magnification) showed the pathological changes in the kidneys of db/db mice. db/m and db/db, normal saline; MET, MET 200 mg/kg; SA 5, SA 5 mg/kg; SA 10, SA 10 mg/kg; SA 20, SA 20 mg/kg. (G) Collagen volume fraction quantified from the stained sections. All data are represented as the mean ± SEM. *p < 0.05, **p < 0.01, and ***p < 0.001 compared with the db/m group; #p < 0.05, ##p < 0.01, and ###p < 0.001 compared with the db/db mice.

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  Figure 5. SA regulated the TGF-β1/Smads signaling in the renal tissue (A) Representative Western blot of TGF-β1 in kidney tissues and quantitative analysis of band intensity. (B) Representative Western blot of Smad2/3 in kidney tissues and quantitative analysis of band intensity. (C) Representative Western blot of Smad7 in kidney tissues and quantitative analysis of band intensity. (D) Representative immunohistochemistry images of TGF-β1 and Smad2/3 in the kidney tissues. db/m and db/db, normal saline; MET, MET 200 mg/kg; SA 5, SA 5 mg/kg; SA 10, SA 10 mg/kg; SA 20, SA 20 mg/kg. (E, F) Mean density of TGF-β1 and Smad2/3. All data are represented as the mean ± SEM (n = 3). *p < 0.05, **p < 0.01, and ***p < 0.001 compared with the db/m group; #p < 0.05, ##p < 0.01, and ###p < 0.001 compared with the db/db mice.

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  Figure 6. Illustration of the derivative synthesis of SA.