Spirolactone provides protection from renal fibrosis by inhibiting the endothelial–mesenchymal transition in isoprenaline-induced heart failure in rats

Figures & data

Table 1 Effect of spironolactone on cardiac functional index

Group	No	LVSP (mmHg)	LVEDP (mmHg)	HR (beats/min)	+dp/dtmax (mmHg/s)	−dp/dtmax (mmHg/s)
Control	8	138±6.2	−5±6.5	428±19.3	9,242±1,945.7	−7,827±1,226.4
Iso	8	108±5.7*	16±5.0*	319±10.5*	3,618±1,201.6*	−2,790±873.0*
Iso + low-dose spironolactone	8	119±11.3**	7±4.7***	341±29.0	5,286±1,274.1**	−4,265±959.8**
Iso + high-dose spironolactone	7	131±10.2***	−2±6.9***	411±17.1***	5,935±1,820.9***	−5,719±1,180.3***

Notes:

* P<0.01 vs control;

** P<0.05,

*** P<0.01 vs Iso. Data shown as mean ± standard deviation.

Abbreviations: LVSP, left ventricle mean systolic pressure; LVEDP, left ventricle end diastolic pressure; HR, heart rate; Iso, isoprenaline; +dp/dt_max, the maximum rate of rise in LV pressure; LV, left ventricle.

Table 2 Effect of spironolactone on ventricular weight and kidney weight in an Iso-induced heart failure rat model

Group	Body weight (g)	LVWI (mg/g)	RVWI (mg/g)	KWI (mg/g)
Control	269±9.9	2.47±0.15	0.62±0.07	4.11±0.12
Iso	228±8.1*	3.25±0.22*	0.84±0.09*	4.75±0.10*
Iso + low-dose spironolactone	242±8.4**	2.89±0.15***	0.75±0.12**	4.52±0.26**
Iso + high-dose spironolactone	248±20.2***	2.63±0.22***	0.68±0.06***	4.34±0.10***

Notes: Data shown as mean ± standard deviation (n=10).

* P<0.01 vs control;

** P<0.05,

*** P<0.01 vs Iso.

Abbreviations: LVWI, left ventricle weight indices; RVWI, right ventricle weight indices; KWI, renal weight indices; SD, standard deviation; Iso, isoprenaline.

Figure 1 Hematoxylin and eosin staining of heart and kidney tissues in rats.

Notes: (A) Heart. (a) Control. (b) Iso. (c) Iso + low-dose spironolactone. (d) Iso + high-dose spironolactone. Magnification: ×200. (B) Kidney. (a) Control. (b) Iso. (c) Iso + low-dose spironolactone. (d) Iso + high-dose spironolactone. Magnification: ×400.
Abbreviation: Iso, isoprenaline.

Figure 2 Collagen areas of heart and kidney tissues in rats.

Notes: (A) Heart. (B) Kidney. (a) Control. (b) Iso. (c) Iso + low-dose spironolactone. (d) Iso + high-dose spironolactone. Magnification: ×200. *P<0.01 vs control; **P<0.01 vs Iso.
Abbreviation: Iso, isoprenaline.

Table 3 Expression of Type I and III collagen in heart and kidney

Group	Heart		Kidney
	Type I collagen (ng/mL)	Type III collagen (ng/mL)	Type I collagen (ng/mL)	Type III collagen (ng/mL)
Control	2.20±0.66	1.98±0.61	2.28±0.39	1.96±0.43
Iso	9.17±0.97*	5.57±0.90*	7.66±1.02*	4.78±0.66*
Iso + low-dose spironolactone	6.11±0.81**	4.06±0.50**	5.32±0.73**	3.19±0.66**
Iso + high-dose spironolactone	4.07±0.81**	3.07±0.76**	3.80±0.49**	2.88±0.37**

Notes: Data are mean ± SD.

* P<0.01 vs control;

** P<0.01 vs Iso.

Abbreviations: SD, standard deviation; Iso, isoprenaline.

Figure 3 Expression of CD31, α-SMA, and TGF-β1 proteins in rats.

Notes: (A) and (B) Immunofluorescence staining of effect of spironolactone in Iso-treated rats. (a) Control. (b) Iso. (c) Iso + low-dose spironolactone. (d) Iso + high-dose spironolactone. CD31 protein (endothelial phenotype) is stained red, α-SMA protein (myofibroblastic phenotype) is green, and nuclei are blue. Magnification: ×200. Western blot analysis of the protein levels (C) and (D) CD31 and α-SMA and (E) TGF-β1. *P<0.01 vs control, **P<0.01 vs Iso, ***P<0.05.
Abbreviations: CD31, cluster of differentiation-31; TGF-β1, transforming growth factor β1; α-SMA, α-smooth muscle actin; Iso, isoprenaline; GAPDH, glyceraldehyde- 3-phosphate dehydrogenase.