Relaxin Reverses Cardiac and Renal Fibrosis in Spontaneously Hypertensive Rats

Abstract

The antifibrotic effects of the peptide hormone relaxin on cardiac and renal fibrosis were studied in 9- to 10-month-old male spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto rats (WKY). Rats (n=8 to 9 per group) were allocated into 3 groups: WKY controls, vehicle-treated SHR (SHR-V), and relaxin-treated SHR (SHR-R). Relaxin (0.5 mg/kg per day) was administered via subcutaneously implanted osmotic mini-pumps over 2 weeks before hearts and kidneys were harvested for analysis. Collagen content was analyzed by hydroxyproline assay, gel electrophoresis, and quantitative histology. Zymography was used to determine matrix metalloproteinase (MMP) expression and Western blotting to determine proliferating cell nuclear antigen (PCNA) expression and α-smooth muscle actin (α-SMA)/myofibroblast expression, whereas cardiac hypertrophy was assessed by myocyte size and real-time polymerase chain reaction of associated genes. The left ventricular (LV) myocardium of SHR-V contained increased collagen levels (by 25±1%, P <0.01 using biochemical analysis and 3-fold; P <0.01 using quantitative histology), enhanced expression of PCNA (by 70±8%; P <0.01), α-SMA (by 32±2%; P <0.05), and the collagen-degrading enzyme MMP-9 (by 70±6%; P <0.05) versus respective levels measured in WKY controls. The kidneys of SHR-V also contained increased collagen (25±2%, P <0.05 using biochemical analysis and 2.4-fold; P <0.01 using quantitative histology). Relaxin treatment significantly normalized collagen content in the LV ( P <0.01) and kidney ( P <0.05), completely inhibited cell proliferation ( P <0.01) and fibroblast differentiation ( P <0.05) in the LV, and increased MMP-2 expression (by 25±1%; P <0.05) without affecting MMP-9 in the LV compared with that measured in SHR-V. Thus, relaxin is a potent antifibrotic hormone with a rapid-occurring efficacy that may have therapeutic potential for hypertensive disease.