Christian Garbar

Cyproheptadine prevents pergolide-induced valvulopathy in rats: an echocardiographic and histopathological study

Serotonergic drugs, such as pergolide, have been associated with the development of cardiac valvular myxoid thickening and regurgitation in humans and more recently in rats. These effects are potentially mediated by the 5-hydroxytryptamine (5-HT)(2B) receptor (5-HT(2B)R). Therefore, we sought to determine whether cyproheptadine, a 5-HT(2B)R antagonist, might prevent toxic valvulopathy in an animal model of pergolide-induced valvular heart disease. For this purpose, 50 male Wistar rats received daily intraperitoneal injections of pergolide (0.5 mg/kg, n = 14), pergolide (0.5 mg/kg) combined with cyproheptadine (10 mg/kg, n = 12), cyproheptadine (10 mg/kg, n = 12), or no injections (control, n = 12) for 20 wk. Echocardiography was performed blindly at baseline and at 10 and 20 wk followed by pathology. At baseline, no differences between groups were found with echocardiography. At 20 wk, aortic regurgitation was present in all pergolide-treated animals, whereas it was less frequently observed in the other groups (P < 0.0001). For the other valves, this difference was less pronounced. On histopathology, not only aortic but also mitral valves were thicker, myxoid, and exhibited more 5-HT(2B)R-positive cells in pergolide-treated animals compared with the other groups. Moreover, regurgitant aortic and mitral valves were thicker than nonregurgitant aortic and mitral valves. In conclusion, we found that cyproheptadine prevented pergolide-induced valvulopathy in rats, which was associated with a reduced number of 5-HT(2B)R-positive valvular cells. This may have important clinical implications for the prevention of serotonergic drug-induced valvular heart disease.

Effect of streptozotocin-induced diabetes on myocardial blood flow reserve assessed by myocardial contrast echocardiography in rats

AbstractThe role of structural and functional abnormalities of small vessels in diabetes cardiomyopathy remains unclear. Myocardial contrast echocardiography allows the quantification of myocardial blood flow at rest and during dipyridamole infusion. The aim of the study was to determine the myocardial blood flow reserve in normal rats compared with Streptozotocin-induced diabetic rats using contrast echocardiography.MethodsWe prospectively studied 40 Wistar rats. Diabetes was induced by intravenous streptozotocin in 20 rats. All rats underwent baseline and stress (dipyridamole: 20 mg/kg) high power intermittent imaging in short axis view under anaesthesia baseline and after six months. Myocardial blood flow was determined and compared at rest and after dipyridamole in both populations. The myocardial blood flow reserve was calculated and compared in the 2 groups. Parameters of left ventricular function were determined from the M-mode tracings and histological examination was performed in all rats at the end of the study.ResultsAt six months, myocardial blood flow reserve was significantly lower in diabetic rats compared to controls (3.09 ± 0.98 vs. 1.28 ± 0.67 ml min-1 g-1; p < 0.05). There were also a significant decrease in left ventricular function and a decreased capillary surface area and diameter at histology in the diabetic group.ConclusionIn this animal study, diabetes induced a functional alteration of the coronary microcirculation, as demonstrated by contrast echocardiography, a decrease in capillary density and of the cardiac systolic function. These findings may offer new insights into the underlying mechanisms of diabetes cardiomyopathy.

Evaluation of contractile function and inotropic reserve with tissue velocity, strain and strain rate imaging in streptozotocin-induced diabetes

AIMS The aim of the present study was to evaluate left ventricular (LV) function and contractile reserve (CR) with Doppler myocardial imaging (DMI) in a small animal model for type 1 diabetes. METHODS AND RESULTS Cardiac function was assessed in anaesthetized Wistar rats 6 and 8 weeks after injection of 60 mg/kg of streptozotocin. At 6 weeks of diabetes, colour DMI echocardiography was performed at rest and during incremental dosages of dobutamine (5, 10, 20 microg/kg/min). Left ventricular fractional shortening was decreased after 8 weeks of follow-up [36 +/- 5 (D) vs. 41 +/- 4% (C); P = 0.049]. After 6 weeks of diabetes, DMI measurements were reduced in the diabetic rats in the inferolateral wall at rest [systolic velocity: 2.5 +/- 0.4 (D) vs. 4.4 +/- 0.3 (C) cm/s; P < 0.001; systolic strain rate: 12.2 +/- 3.4 (D) vs. 17.4 +/- 3.2 (C) 1/s; P = 0.012] and during inotropic stimulation [delta velocity (cm/s): 0.2 +/- 0.1 (D) vs. 0.5 +/- 0.3 (C)/5 microg dobutamine; P = 0.002; delta strain rate (1/s): 1.4 +/- 0.9 (D) vs. 3.3 +/- 2.2 (C)/5 microg dobutamine; P = 0.049]. Furthermore, the intraventricular delay in time-to-peak systolic strain was larger in diabetes [20 +/- 18 (D) vs. 10 +/- 7 (C) ms; P= 0.01]. Systolic mitral annular velocity was also lower in the diabetic rats at rest [2.7 +/- 0.4 (D) vs. 3.5 +/- 0.4 (C) cm/s; P < 0.001] and in response to dobutamine [delta velocity (cm/s): 0.1 +/- 0.1 (D) vs. 0.3 +/- 0.2 (C)/5 microg dobutamine; P = 0.013]. CONCLUSION In experimental diabetes, a reduction in radial and longitudinal LV function and CR can be detected with DMI before the onset of a reduced global LV function.

Echocardiographic and histological assessment of age-related valvular changes in normal rats.

Aging is associated with morphologic and functional alterations of the rats left ventricle. However, the time-course of valvular function and morphology in normal aging rats has not yet been studied. For this purpose, 30 male Wistar rats (318 +/- 5g, 10 weeks old) underwent serial echocardiograms for 58 weeks under sodium pentobarbital 50 mg/kg IP anesthetization followed by necropsy. Histopathology was also performed in two additional groups of 10 rats at 10 and 30 weeks of age. Regurgitations were considered as any retrograde flow on 2-D or M-mode color Doppler echocardiography. Tricuspid regurgitation was already found at 10 weeks of age and became more frequent with age. Pulmonary, mitral and aortic regurgitation was seldom observed at 10 weeks but became more frequent after 30 weeks. For the mitral and aortic valve, this was also associated with an increase in valvular thickness because of nodular or segmental myxoid leaflet changes. The severity of valvular regurgitations did not increase with age. In conclusion, aging leads to morphologic and functional valvular changes in normal rats. This is important when investigating models of valvular heart disease in small animals.

EFFECT OF HIGH-INTENSITY ULTRASOUND-TARGETED MICROBUBBLE DESTRUCTION ON PERFUSION AND FUNCTION OF THE RAT HEART ASSESSED BY PINHOLE-GATED SPECT

Although ultrasound-targeted microbubble destruction (UTMD) has been shown to induce bioeffects, UTMD is still desirable for therapeutic applications. Therefore, we studied the effects of UTMD on perfusion and function of the rat heart, assessed by (99m)Tc-MIBI pinhole-gated SPECT (Ph-gSPECT) compared with biomarker release and histopathology. Fifty-two male Wistar rats were studied. UTMD was performed using SonoVue, with a mechanical index of 1.0 or 1.6. Controls were treated without microbubbles or without ultrasound application. At baseline, day 1, day 7 and day 30, 35 rats were imaged with (99m)Tc-MIBI Ph-gSPECT to quantify left ventricular perfusion and function. In addition, troponin release and histopathology were investigated. No significant differences were observed for left ventricular ejection fractions, end-systolic and end-diastolic volumes, regional perfusion and functional scores up to 30 days after UTMD compared with controls. UTMD induced mild troponin release and early erythrocyte extravasation without necrosis, inflammation or fibrosis. Although UTMD has the potential to induce microlesions of the heart in small animals, these effects were transient without histological evidence of irreversible damage. Furthermore, UTMD does not induce abnormalities on perfusion or function of the heart, as assessed by Ph-gSPECT, which is reassuring concerning the use of SonoVue for potential therapeutic applications. (E-mail: sophie.hernot@gmail.com).