Effects of cardiac contractility modulation on the autophagy of cardiomyocytes in rabbits with chronic heart failure and the underlying mechanisms

doi:10.3969/j.issn.1004-583X.2024.11.011

Abstract

Abstract:

Objective To explore the effect of cardiac contractility modulation (CCM) on the autophagy of cardiomyocytes in rabbits with chronic heart failure (CHF) and the underlying mechanisms. Methods Thirty healthy New Zealand White rabbits weighing 2.5-3.5 kg were randomly assigned to the sham operation group, HF group, and CCM group, with 10 rabbits in each group. A CHF model was established by the ascending aortic constriction method, followed by CCM for 4 weeks. At 12 and 16 weeks, echocardiography was employed to assess the cardiac function. Expression level of the microtuble-associated protein 1 light chain 3 (LC3) was detected by immunofluorescence staining. Protein levels of P62, Beclin1, light chain 3B II/I (LC3B II/I), protein kinase B (AKT)1, AKT2, AKT3, phosphatidylinositol 3-kinase (PI3K) α110, PI3K α85 and mammalian target of rapamycin (mTOR) in myocardial tissue were detected by Western blot.Results At 12 weeks, rabbits in the HF and CCM groups had significantly larger left ventricular end-diastolic diameter (LVEDD) and left ventricular end-systolic diameter (LVESD), and lower left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) than those of sham operation group (P<0.05). At 16 weeks, significant improvements in LVEF, LVFS, LVESD and LVEDD were observed in the CCM group than HF group (P<0.05). Compared with those of the HF group, rabbits in the CCM group had significantly upregulated p62, PI3K α110 and PI3K α85, less LC3-positive immunofluorescence staining, downregulated Beclin1, LC3B, AKT1-3 and mTOR in myocardium and lower LC3B(II/I) ratio (all P<0.05). Conclusion CCM can improve the abnormal expression of LC3B (II/I), Beclin1 and p62 protein in myocardium of CHF rabbits, reduce the activity of autophagy in rabbit myocardium, and improve myocardial contractile and diastolic function by regulating the PI3K/AKT signaling pathway.

Key words: heart failure, autophagy, myocardial contraction, autophagy related protein, Pi3k/akt, rabbit

CLC Number:

R541.6

Hao Qingqing, Lv Shilin, Zhang Jing, Zhang Litao, Zhang Feifei. Effects of cardiac contractility modulation on the autophagy of cardiomyocytes in rabbits with chronic heart failure and the underlying mechanisms[J]. Clinical Focus, 2024, 39(11): 1026-1034.

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URL: https://huicui.hebmu.edu.cn/EN/10.3969/j.issn.1004-583X.2024.11.011

https://huicui.hebmu.edu.cn/EN/Y2024/V39/I11/1026

Figures/Tables 11

Tab.1 Echocardiographic results in three groups before thoracotomy

组别	例数	IVSd(mm)	LVPWd(mm)	LVESD(mm)	LVEDD(mm)	LVEF (%)	LVFS (%)
Sham组	9	2.04±0.18	1.98±0.15	9.07±0.57	11.31±0.26	67.32±3.62	34.39±3.04
HF组	9	2.01±0.20	2.00±0.15	9.20±0.44	11.17±0.47	66.94±3.49	34.40±3.35
CCM组	9	203.00±0.20	1.98±0.15	9.08±0.57	10.98±0.51	68.33±2.51	35.54±2.43
$F$ 值		0.06	0.04	0.17	0.34	0.44	0.45
P值		0.942	0.963	0.841	0.280	0.649	0.642

Tab.1 Echocardiographic results in three groups before thoracotomy

组别	例数	IVSd(mm)	LVPWd(mm)	LVESD(mm)	LVEDD(mm)	LVEF (%)	LVFS (%)
Sham组	9	2.04±0.18	1.98±0.15	9.07±0.57	11.31±0.26	67.32±3.62	34.39±3.04
HF组	9	2.01±0.20	2.00±0.15	9.20±0.44	11.17±0.47	66.94±3.49	34.40±3.35
CCM组	9	203.00±0.20	1.98±0.15	9.08±0.57	10.98±0.51	68.33±2.51	35.54±2.43
$F$ 值		0.06	0.04	0.17	0.34	0.44	0.45
P值		0.942	0.963	0.841	0.280	0.649	0.642

Tab.2 Echocardiographic results of three groups at 12 weeks postoperatively

组别	例数	IVSd(mm)	LVPWd(mm)	LVESD(mm)	LVEDD(mm)	LVEF (%)	LVFS (%)
Sham组	9	2.09±0.23	2.05±0.19	9.07±0.32	11.18±0.56	67.74±4.81	34.18±3.47
HF组	9	2.01±0.24	2.09±0.15	12.13±0.52^*	14.70±0.^*	38.22±1.99^*	19.58±1.82^*
CCM组	9	2.05±0.14	2.13±0.18	11.98±0.54^*	14.48±0.78^*	38.11±1.62^*	19.64±1.43^*
$F$ 值		0.35	0.40	120.66	68.55	265.39	109.87
P值		0.708	0.678	0.000	0.000	0.000	0.000

Tab.2 Echocardiographic results of three groups at 12 weeks postoperatively

组别	例数	IVSd(mm)	LVPWd(mm)	LVESD(mm)	LVEDD(mm)	LVEF (%)	LVFS (%)
Sham组	9	2.09±0.23	2.05±0.19	9.07±0.32	11.18±0.56	67.74±4.81	34.18±3.47
HF组	9	2.01±0.24	2.09±0.15	12.13±0.52^*	14.70±0.^*	38.22±1.99^*	19.58±1.82^*
CCM组	9	2.05±0.14	2.13±0.18	11.98±0.54^*	14.48±0.78^*	38.11±1.62^*	19.64±1.43^*
$F$ 值		0.35	0.40	120.66	68.55	265.39	109.87
P值		0.708	0.678	0.000	0.000	0.000	0.000

Tab.3 Echocardiographic results of three groups at 16 weeks postoperatively

组别	例数	IVSd(mm)	LVPWd(mm)	LVESD(mm)	LVEDD(mm)	LVEF (%)	LVFS (%)
Sham组	9	2.06±0.20	2.03±0.19	9.40±0.44	11.29±0.42	68.00±4.47	34.31±3.66
HF组	9	2.05±0.25	2.11±0.20	11.82±0.36^*	14.40±0.38^*	37.11±1.69^*	18.76±1.18^*
CCM组	9	2.03±0.17	2.13±0.21	10.01±0.28^*#	12.94±0.48^*#	45.22±1.86^*#	22.72±1.15^*#
$F$ 值		0.04	0.63	106.52	116.98	263.75	109.73
P值		0.965	0.544	0.000	0.000	0.000	0.000

Tab.3 Echocardiographic results of three groups at 16 weeks postoperatively

组别	例数	IVSd(mm)	LVPWd(mm)	LVESD(mm)	LVEDD(mm)	LVEF (%)	LVFS (%)
Sham组	9	2.06±0.20	2.03±0.19	9.40±0.44	11.29±0.42	68.00±4.47	34.31±3.66
HF组	9	2.05±0.25	2.11±0.20	11.82±0.36^*	14.40±0.38^*	37.11±1.69^*	18.76±1.18^*
CCM组	9	2.03±0.17	2.13±0.21	10.01±0.28^*#	12.94±0.48^*#	45.22±1.86^*#	22.72±1.15^*#
$F$ 值		0.04	0.63	106.52	116.98	263.75	109.73
P值		0.965	0.544	0.000	0.000	0.000	0.000

Fig.1 The typical echocardiographic imaging of HF rabbits and the echocardiographic parameters of the three groups a-b. Typical echocardiographic imaging of HF rabbits; c-h. Comparison of echocardiographic parameters among the three groups

Fig.2 LC3 immunofluorescence staining of myocardial tissue among three groups (×400)

Tab.4 Beclin1, P62 and LC3B(II/I) protein expression levels of myocardial tissue among three groups

组别	例数	Beclin1	P62	LC3B(II/I)
Sham组	9	0.34±0.15	0.50±0.08	1.00±0.10
HF组	9	0.85±0.06^*	0.18±0.07^*	1.47±0.15^*
CCM组	9	0.60±0.11^*#	0.28±0.06^*#	1.13±0.12^#

Fig.3 The protein bands and expression levels of Beclin1, P62 and LC3B(II/I) of myocardial tissue among the three groups a. Western blot results of Beclin1, P62 and LC3B(II/I) proteins in the three groups; b. Beclin1 protein expression; c.P62 protein expression; d. LC3B (II/I) expression

Fig.4 The protein bands and expression levels of AKT1, AKT2, AKT3, PI3K(α110) and PI3K(α85) of myocardial tissue among the three groups a. Western blot results of AKT1, AKT2, AKT3, PI3K(α110) and PI3K(α85) in the three groups; b.AKT1 expression; c. AKT2 expression; d.AKT3 expression; e.PI3K(α110) protein expression; f.PI3K (α85) protein expression

Tab.5 AKT1, AKT2, AKT3, PI3K (α110), and PI3K (α85) protein expression levels of myocardial tissue among three groups

组别	例数	AKT1	AKT2	AKT3	PI3K (α110)	PI3K (α85)
Sham组	9	0.10±0.03	0.24±0.07	0.19±0.05	0.40±0.04	0.62±0.10
HF组	9	0.52±0.06^*	0.58±0.07^*	0.50±0.06^*	0.15±0.05^*	0.24±0.04^*
CCM组	9	0.22±0.04^*#	0.42±0.06^*#	0.29±0.06^*#	0.26±0.08^*#	0.45±0.05^*#

Fig.5 The protein bands and expression levels of mTOR of myocardial tissue among the three groups

Tab.6 The mTOR protein expression levels of myocardial tissue among three groups

组别	例数	mTOR
Sham组	9	0.35±0.14
HF组	9	0.75±0.22^*
CCM组	9	0.50±0.14^#

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