Objective To investigate the intervention effect of sacubitril/valsartan(Sac/Val)on doxorubicin-induced heart failure(HF)in rabbits and its regulative effect on the transforming growth factor β1(TGF-β1)/SMAD family member 3(Smad3)/connective tissue growth factor(CTGF)signaling pathway.Methods Thirty-eight male New Zealand rabbits were subjected,and 8 of them were randomly assigned into a control group.The other 30 rabbits were injected with doxorubicin to establish a rabbit HF model,and finally,there were 6 rabbits in a model group,7 in a valsartan group,and 7 in a Sac/Val group.After 8 weeks of intervention,echocardiography[left ventricular ejection fraction(LVEF),left ventricular fractional shortening(LVFS),left ventricular end-diastolic diameter(LVEDD),left ventricular end-systolic diameter(LVESD)],and myocardial histopathologic observation(HE staining,Masson staining)were performed,and collogen volume fraction(CVF)was calculated.The levels of N-terminal pro-B-type natriuretic peptide(NT-proBNP),soluble growth stimulation expressed gene 2(sST2),galectin-3(Gal-3)were detected.Activities of renin-angiotensin-aldosterone system(RAAS)and levels of atrial natriuretic peptide,B-type brain natriuretic peptide(BNP),cyclic guanosine monophosphate(cGMP)and protein kinase G(PKG)were measured.The expression of α-smooth muscle actin(α-SMA),typeⅠ collagen,TGF-β1,Smad3,recombinant SMAD family member 7(Smad7)and CTGF in the myocardial tissues were detected.Results Compared with the control group,the model group exhibited significantly lower LVEF and LVFS and decreased expression of Smad7,higher LVEDD and LVESD(P＜0.01).The CVF of each group was(7.15±0.82)％、(43.20±5.09)％、(29.53±4.05)％、(22.48±2.93)％.Valsartan and Sac/Val treatment resulted in obvious increases in LVEF and LVFS,up-regulation of Smad7,decreased in LVEDD,LVESD and CVF,reduced levels of NT-proBNP,sST2,Gal-3,AngⅡ,aldosterone,atrial natriuretic peptide,BNP,cGMP and PKG,and down-regulation of α-SMA,collagen I,TGF-β1,Smad3 and CTGF when compared with the model group(P＜0.01).Sac/Val treatment showed better effects in above indicators than simple valsartan treatment(P＜0.01).Conclusion Sac/Val can reduce myocardial fibrosis and improve cardiac function in doxorubicin-induced HF rabbits,which may be related to the dual inhibition of TGF-β1/Smad3/CTGF signaling pathway by upregulating atrial natriuretic peptide and BNP levels and blocking RAAS activation.

Objective To study the protective effects and mechanism of sacubitril(Sac)/valsartan(Val)on cardiomyocytes of rabbits with heart failure induced by doxorubicin(DOX).Methods Thirty New Zealand rabbits were selected to establish DOX-induced heart failure rabbit model.Twenty-five rabbits with successful modeling were randomly assigned to model group(DOX group,n=9),DOX+Val group(n=8),and DOX+Sac/Val group(n=8);and another 8 New Zealand rabbits were selected as blank group.The DOX+Val group was gavaged with 4.65 mg/kg Val suspension each time,the DOX+Sac/Val group was gavaged with 9.3 mg/kg Sac/Val suspension each time,and the blank group and DOX group were gavaged with equal volume of distilled water each time.Each group was gavaged twice a day for 8 weeks.After 8 weeks of administration,echocardiography was used to measure left ventricular end-diastolic diameter(LVDD),left ventricular end-systolic diameter(LVSD),left ventricular ejection fraction(LVEF),and left ventricular fractional shortening(LVFS).The heart mass index(HMI)and left ventricular mass index(LVMI)were calculated.The pathological morphology and myocardial fibrosis of myocardial tissue were observed by hematoxylin-eosin(H-E)and Masson staining.The ultrastructure of cardiomyocytes was observed by transmission electron microscope.Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)staining was used to observe cardiomyocytes apoptosis and apoptosis rate was calculated.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of N-terminal pro-brain natriuretic peptide(NT-proBNP),high-sensitivity cardiac troponin I(Hs-cTNI),angiotensin Ⅱ(Ang Ⅱ),aldosterone(ALD),atrial natriuretic peptide(ANP),brain natriuretic peptide(BNP),cyclic guanosine monophosphate(cGMP),and protein kinase G(PKG)in serum.Quantitative polymerase chain reaction(qPCR)was used to detect the expression of natriuretic peptide receptor A(NPR-A),cGMP-specific phosphodiesterase 5A(PDE5A[cGMP]),PKG,B-cell lymphoma 2(Bcl-2),Bcl-2 associated X protein(Bax),and cysteine aspartate protease 3(caspase 3)mRNA in myocardial tissue.Western blotting was used to detect the expression of phosphorylated cAMP response element-binding protein(p-CREB),phosphorylated Bcl-2 related death promoting factor(p-Bad),Bcl-2,Bax,and caspase 3 proteins in myocardial tissue.Results Compared with the blank group,the LVDD and LVSD in the DOX group were increased(both P<0.01),the LVEF and LVFS were decreased(both P<0.01)and the HMI and LVMI were increased(both P<0.01);the apoptosis and apoptosis rate of cardiomyocytes were increased(P<0.01);the levels of NT-proBNP,Hs-cTNI,Ang Ⅱ,ALD,ANP,BNP,cGMP and PKG and the expression of NPR-A,PDE5A(cGMP),PKG,p-CREB,Bax and caspase 3 were all increased(all P<0.01),while the expression of Bcl-2 was decreased(P<0.01),and the expression of p-Bad had no significant difference(P>0.05).Compared with the DOX group,the LVDD and LVSD of the DOX+Sac/Val group and DOX+Val group were decreased(all P<0.01),the LVEF and LVFS were increased(all P<0.01)and the HMI and the LVMI were decreased(all P<0.01);the apoptosis and apoptosis rate of cardiomyocytes were decreased(all P<0.01);the levels of NT-proBNP,Hs-cTNI,Ang Ⅱ,ALD,ANP,BNP,cGMP and PKG and the expression of NPR-A,PDE5A(cGMP),PKG,Bax and caspase 3 were all decreased(all P<0.01),while the expression of Bcl-2 was increased(P<0.01);and the expression of p-CREB and p-Bad was increased in the DOX+Sac/Val group(both P<0.01),but there was no significant difference in the DOX+Val group(both P>0.05).Compared with the DOX+Val group,the DOX+Sac/Val group showed a decrease in all indicators except for LVEF,LVFS,NPR-A,ANP,BNP,cGMP,PDE5A(cGMP),PKG,p-CREB,p-Bad,and Bcl-2,which were all elevated(all P<0.05).Myocardial pathology and transmission electron microscopy showed that Sac/Val effectively protected cardiomyocytes,reduced cardiomyocytes apoptosis and myocardial fibrosis,and these effects were significantly better than those of Val.Conclusion Sac/Val can effectively reduce cardiomyocytes apoptosis,improve cardiac function and reduce myocardial fibrosis in rabbits with heart failure,and these effects are superior to Val.Its mechanism may be related to activating the NPR-A/cGMP/PKG signaling pathway and inhibiting renin-angiotensin-aldosterone system.

Objective To investigate the intervention effect of sacubitril/valsartan(Sac/Val)on doxorubicin-induced heart failure(HF)in rabbits and its regulative effect on the transforming growth factor β1(TGF-β1)/SMAD family member 3(Smad3)/connective tissue growth factor(CTGF)signaling pathway.Methods Thirty-eight male New Zealand rabbits were subjected,and 8 of them were randomly assigned into a control group.The other 30 rabbits were injected with doxorubicin to establish a rabbit HF model,and finally,there were 6 rabbits in a model group,7 in a valsartan group,and 7 in a Sac/Val group.After 8 weeks of intervention,echocardiography[left ventricular ejection fraction(LVEF),left ventricular fractional shortening(LVFS),left ventricular end-diastolic diameter(LVEDD),left ventricular end-systolic diameter(LVESD)],and myocardial histopathologic observation(HE staining,Masson staining)were performed,and collogen volume fraction(CVF)was calculated.The levels of N-terminal pro-B-type natriuretic peptide(NT-proBNP),soluble growth stimulation expressed gene 2(sST2),galectin-3(Gal-3)were detected.Activities of renin-angiotensin-aldosterone system(RAAS)and levels of atrial natriuretic peptide,B-type brain natriuretic peptide(BNP),cyclic guanosine monophosphate(cGMP)and protein kinase G(PKG)were measured.The expression of α-smooth muscle actin(α-SMA),typeⅠ collagen,TGF-β1,Smad3,recombinant SMAD family member 7(Smad7)and CTGF in the myocardial tissues were detected.Results Compared with the control group,the model group exhibited significantly lower LVEF and LVFS and decreased expression of Smad7,higher LVEDD and LVESD(P＜0.01).The CVF of each group was(7.15±0.82)％、(43.20±5.09)％、(29.53±4.05)％、(22.48±2.93)％.Valsartan and Sac/Val treatment resulted in obvious increases in LVEF and LVFS,up-regulation of Smad7,decreased in LVEDD,LVESD and CVF,reduced levels of NT-proBNP,sST2,Gal-3,AngⅡ,aldosterone,atrial natriuretic peptide,BNP,cGMP and PKG,and down-regulation of α-SMA,collagen I,TGF-β1,Smad3 and CTGF when compared with the model group(P＜0.01).Sac/Val treatment showed better effects in above indicators than simple valsartan treatment(P＜0.01).Conclusion Sac/Val can reduce myocardial fibrosis and improve cardiac function in doxorubicin-induced HF rabbits,which may be related to the dual inhibition of TGF-β1/Smad3/CTGF signaling pathway by upregulating atrial natriuretic peptide and BNP levels and blocking RAAS activation.

Objective: To observe the protective effect of salidroside pretreatment on rabbit heart after limb ischemia/reperfusion (I/R). Methods: A total of 24 New Zealand rabbits were randomly and equally divided into sham operation group, I/R + placebo group （I/R group）and salidroside pretreatment group（salidroside group）. Before establishment of rabbit models of limb I/R, salidroside group received salidroside injection via ear marginal vein, and I/R group received saline injection, once a day for three days. After model establishment, echocardiography was used to evaluate rabbit cardiac function of each group 4h after reperfusion, including left ventricular end-systolic dimension (LVESd), left ventricular end-diastolic dimension (LVEDd), left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (FS). Blood sample was taken to measure serum cardiac troponin I (cTnI) and tumor necrosis factor (TNF)-α. Some ventricular tissue homogenates were taken to measure levels of superoxide dismutase (SOD) and malonyl diadehyde (MDA). Then heart sample was taken to receive pathological examination. Results: Compared with I/R group 4h after reperfusion, there were significant decrease in LVESd [(0.69±0.07) mm vs. (0.62±0.05) mm] and significant increase in LVEF [(64.6±3.4) % vs. (72.1±3.6) %], FS [(34.2±3.2) % vs. (41.7±3.4) %] (P<0.05 all), but these indexes of salidroside group were all no significant different than those of sham operation group (P>0.05). Compared with I/R group, there were significant decrease in cTnI [(5.24±0.34) μg/ml vs. (1.06±0.12) μg/ml], MDA [(8.92±2.18) μmol/L vs. (6.79±1.43) μmol/L] and TNF-α [(37.43±10.02) pg/ L vs. (19.73±6.31) pg/ L], and significant increase in SOD level [(16.61±3.75) U/ml vs. (22.26±4.73) U/ml] in salidroside group (P<0.05 all). Pathological results indicated that injury degree in salidroside group was significantly attenuated than that of I/R group. Conclusion: Salidroside pretreatment could protect cardiac function and relieve rabbit cardiac injury after limb ischemia/reperfusion.