Pyridostigmine Treatment Significantly Alleviates Isoprenaline-Induced Chronic Heart Failure in Rats

Figures

• 

  Figure 1. Effects of pyridostigmine treatment on acetylcholinesterase (AChE) activity. Control—0.9% NaCl s.c. for 7 days + 28 days observation + tap water p.o. 14 days; Pyr—0.9% NaCl s.c. for 7 days + 28 days observation + 22 mg/kg/day of pyridostigmine p.o. 14 days; Iso—5 mg/kg/day of isoprenaline s.c. for 7 days + 28 days observation + tap water p.o. 14 days; Iso + Pyr—5 mg/kg/day of isoprenaline s.c. for 7 days + 28 days observation + 22 mg/kg/day of pyridostigmine p.o. 14 days; ** p < 0.01 versus Control; *** p < 0.001 versus Control; †† p < 0.01 versus Iso; ††† p < 0.001 versus Iso.

• 

  Figure 2. Effects of pyridostigmine treatment on echocardiogram (ECHO) parameters. (a) IVSd—interventricular septum thickness end diastole; (b) IVSs—interventricular septum thickness end systole; (c) LVIDd—left ventricle internal diameter end diastole; (d) LVIDs—left ventricle internal diameter end systole; (e) PWDd—posterior wall diameter end diastole; (f) PWDs—posterior wall diameter end systole; (g) EDV—end-diastolic volume; (h) ESV—end-systolic volume; (i) SV—stroke volume; (j) EF—ejection fraction; ∆IVSd/IVSs—interventricular septum thickness end diastole and end systole change; ∆LVIDd/LVIDs—left ventricle internal diameter end diastole and end systole change; ∆PWDd/PWDs—posterior wall diameter end diastole and end systole change; ∆EDV—end-diastolic volume change; ∆ESV—end-systolic volume change; ∆SV—stroke volume change; ∆EF—ejection fraction change; 2—between first and second recording, after CHF induction; 3—between second and third recording, after treatment; Control—0.9% NaCl s.c. for 7 days + 28 days observation + tap water p.o. 14 days; Pyr—0.9% NaCl s.c. for 7 days + 28 days observation + 22 mg/kg/day of pyridostigmine p.o. 14 days; Iso—5 mg/kg/day of isoprenaline s.c. for 7…

• 

  Figure 3. Effects of pyridostigmine treatment on prooxidative and antioxidative markers in myocardial tissue samples. (a) Thiobarbituric acid reactive substances (TBARS); (b) nitrite levels (NO2−); (c) superoxide dismutase (SOD); (d) reduced glutathione (GSH); (e) catalase (CAT); Control—0.9% NaCl s.c. for 7 days + 28 days observation + tap water p.o. 14 days; Pyr—0.9% NaCl s.c. for 7 days + 28 days observation + 22 mg/kg/day of pyridostigmine p.o. 14 days; Iso—5 mg/kg/day of isoprenaline s.c. for 7 days + 28 days observation + tap water p.o. 14 days; Iso + Pyr—5 mg/kg/day of isoprenaline s.c. for 7 days + 28 days observation + 22 mg/kg/day of pyridostigmine p.o. 14 days; * p < 0.05 versus Control; ** p < 0.01 versus Control; *** p < 0.001 versus Control; †† p < 0.01 versus Pyr, ††† p < 0.001 versus Pyr, ‡‡ p < 0.01 versus Iso, and ‡‡‡ p < 0.001 versus Iso.

• 

  Figure 4. Effects of pyridostigmine treatment on histological structure of cardiac muscle, septal region of the heart, H&E staining magnification ×200, scale bar 50 µm. (a) Control—normal histological structure of cardiac muscle; (b) Iso—extensive myocardial fibrosis (asterisk), cardiomyocyte damage (black arrow), and erythrocyte extravasation in the endomysium (blue arrow); (c) Pyr—mild hypertrophy of cardiomyocytes is noted (black arrow), with erythrocyte extravasation in the endomysium (blue arrow); (d) Iso + Pyr—largely preserved tissue architecture, with an increased number of inflammatory cells and fibroblasts observed in the endomysium (black arrow). (e) Effects of pyridostigmine treatment on the tissue damage score. Control—0.9% NaCl s.c. for 7 days + 28 days observation + tap water p.o. 14 days; Pyr—0.9% NaCl s.c. for 7 days + 28 days observation + 22 mg/kg/day of pyridostigmine p.o. 14 days; Iso—5 mg/kg/day of isoprenaline s.c. for 7 days + 28 days observation + tap water p.o. 14 days; Iso + Pyr—5 mg/kg/day of isoprenaline s.c. for 7 days + 28 days observation + 22 mg/kg/day of pyridostigmine p.o. 14 days. *** p < 0.001 versus Control; ††† p < 0.001 versus Pyr; ‡‡‡ p < 0.001…

• 

  Figure 5. Effects of pyridostigmine treatment on histological structure of cardiac muscle, septal region of the heart, Masson’s trichrome stain, magnification ×200, scale bar 50 µm. (a) Control—representative image showing the normal histological structure of cardiac muscle. Cardiomyocytes are stained red; (b) Iso—extensive myocardial fibrosis is observed, with fibrotic areas stained blue (asterisk). Cardiomyocyte damage is indicated (red arrow), along with erythrocyte extravasation in the endomysium (black arrow); (c) Pyr—cardiomyocytes are stained red, with mild hypertrophy noted (red arrow). Erythrocyte extravasation is present in the endomysium (black arrow); (d) Iso + Pyr—largely preserved tissue architecture with erythrocyte extravasation in the endomysium (black arrow). Fibrosis is present in a small area, with collagen fibres stained blue (asterisk). Dilated capillaries are visible in the endomysium. Control—0.9% NaCl s.c. for 7 days + 28 days observation + tap water p.o. 14 days; Pyr—0.9% NaCl s.c. for 7 days + 28 days observation + 22 mg/kg/day pyridostigmine p.o. 14 days; Iso—5 mg/kg/day isoprenaline s.c. for 7 days + 28 days observation + tap water p.o. 14 days; Iso + Pyr—5…

• 

  Figure 6. Study design. Control group (Control; n = 6); pyridostigmine group (Pyr; n = 6); isoprenaline group (Iso; n = 8); pyridostigmine group (Iso + Pyr; n = 8); NS—normal saline; ISO—isoprenaline; TW—tap water; PYR—pyridostigmine; CHF—chronic heart failure.