DACT1 Involvement in the Cytoskeletal Arrangement of Cardiomyocytes in Atrial Fibrillation by Regulating Cx43

Hou, Jian; Yue, Yuan; Hu, Bo; Xu, Guangtao; Su, Ruibing; Lv, Linhua; Huang, Jiaxing; Yao, Jianping; Guan, Yuanjun; Wang, Keke; Wu, Zhongkai

doi:10.21470/1678-9741-2019-0033

Jian Hou

MD

Department of Cardiac Surgery, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, GD, People’s Republic of China.

NHC Key Laboratory of Assisted Circulation, Sun Yat-Sen University, Guangzhou, GD, People’s Republic of China.

# J. Hou, Y. Yue, and B. Hu contributed equally to this study.

Yuan Yue

MD

Department of Cardiac Surgery, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, GD, People’s Republic of China.

NHC Key Laboratory of Assisted Circulation, Sun Yat-Sen University, Guangzhou, GD, People’s Republic of China.

# J. Hou, Y. Yue, and B. Hu contributed equally to this study.

Bo Hu

MD

Department of Pathology and Molecular Medicine Center, Jiaxing Hospital of Traditional Chinese Medicine, Jiaxing University, Jiaxing, ZJ, People’s Republic of China.

# J. Hou, Y. Yue, and B. Hu contributed equally to this study.

Guangtao Xu

MD

Department of Pathology and Molecular Medicine Center, Jiaxing Hospital of Traditional Chinese Medicine, Jiaxing University, Jiaxing, ZJ, People’s Republic of China.

Ruibing Su

MD

Department of Forensic Pathology, Shantou University Medical College, Shantou, GD, People’s Republic of China.

Linhua Lv

MD

Department of Cardiac Surgery, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, GD, People’s Republic of China.

Jiaxing Huang

MD

Department of Cardiac Surgery, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, GD, People’s Republic of China.

Jianping Yao

MD

Department of Cardiac Surgery, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, GD, People’s Republic of China.

Yuanjun Guan

MD

Core Lab Plat for Medical Science, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, GD, People’s Republic of China.

Keke Wang

MD

Department of Cardiac Surgery, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, GD, People’s Republic of China.

NHC Key Laboratory of Assisted Circulation, Sun Yat-Sen University, Guangzhou, GD, People’s Republic of China.

Zhongkai Wu Correspondence Address: Zhongkai Wu, Department of Cardiac Surgery, The First Affiliated Hospital of Sun Yat-Sen University, 58 Zhongshan 2nd Rd., Guangzhou, GD, People’s Republic of China, Zip Code: 510080. E-mail: wuzhk@mail.sysu.edu.cn

MD

Department of Cardiac Surgery, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, GD, People’s Republic of China.

NHC Key Laboratory of Assisted Circulation, Sun Yat-Sen University, Guangzhou, GD, People’s Republic of China.

http://orcid.org/0000-0003-1184-1182

Correspondence Address: Zhongkai Wu, Department of Cardiac Surgery, The First Affiliated Hospital of Sun Yat-Sen University, 58 Zhongshan 2nd Rd., Guangzhou, GD, People’s Republic of China, Zip Code: 510080. E-mail: wuzhk@mail.sysu.edu.cn

#
J. Hou, Y. Yue, and B. Hu contributed equally to this study.

No conflict of interest.

Abbreviations, acronyms & symbols
AF	= Atrial fibrillation	LA	= Left atrial
AI	= Aortic insufficiency	LVD	= Left ventricular diastolic
AR	= Aortic regurgitation	LVPW	= Left ventricular posterior wall
AS	= Aortic stenosis	LVS	= Left ventricular systolic
B	= Beta-blockers	M	= Male
cDNA	= Complementary deoxyribonucleic acid	MOI	= Multiplicity of infection
Cx43	= Connexin 43	MI	= Mitral insufficiency
D	= Digoxin	MS	= Mitral stenosis
DACT1	= Dishevelled binding antagonist of beta catenin 1	N	= Nitrates
DAPI	= 2-(4-Amidinophenyl)-6-indolecarbamidine dihydrochloride	NYHA	= New York Heart Association
DI	= Diuretics	PBS	= Phosphate-buffered saline
DMEM	= Dulbecco's Modified Eagle Medium	PFA	= Paraformaldehyde
ECG	= Electrocardiogram	PVDF	= Polyvinylidene difluoride
EF	= Ejection fraction	RA	= Right atrial
F	= Female	RV	= Right ventricular
F-Actin	= Fibrous actin	SACs	= Stretch-activated channels
FBS	= Fetal bovine serum	SDS-PAGE	= Sodium dodecyl sulfate polyacrylamide gel electrophoresis
GAPDH	= Glyceraldehyde-3-phosphate dehydrogenase	SPSS	= Statistical Package for the Social Sciences
GFP	= Green fluorescent protein	SR	= Sinus rhythm
GSK3-β	= Glycogen synthase kinase beta	TI	= Tricuspid insufficiency
ICa,L	= L-type Ca²⁺ current	TS	= Tricuspid stenosis
IVS	= Interventricular septum

Age	Sex	VD	LA (mm)	LVD (mm)	LVS (mm)	IVS (mm)	LVPW (mm)	RA (mm)	RV (mm)	EF (%)	NYHA	Drug therapy	CHADS2 score	CHA2DVASc
SR (n=10)
31	M	MS, MI, AI	50	51	30	9	9	46	25	75	2	D, DI	0	0
66	M	MS, MI, TI, AR, AS	36	52	35	11	10	45	52	62	2	A, D, DI	1	1
66	F	MI, AS, AI	37	52	36	13	10	43	19	58	2	D, DI	1	2
51	M	MI, AI	37	58	34	9	10	45	22	70	2	A, D, DI	1	2
43	F	MS, MI, TI, AS, AI	51	46	31	8	8	49	18	62	2	D, DI	0	1
65	M	MS, MI, TS, TI, AI	47	45	30	9	9	70	25	63	3	A, D, DI	2	2
57	M	MI, AS, AI	31	69	50	14	13	36	20	53	3	A, D, DI, N	0	0
71	F	MI, AS, AI	38	53	37	14	12	41	19	55	2	D, DI	1	2
37	F	TI	45	35	22	8	11	46	17	68	2	D, DI	2	3
58	M	MS, MI, TI, AI	55	51	31	8	8	51	24	66	2	D, DI, B	0	0
TOTAL
54.5±13.618	6M/4F		42.7±7.931	51.2±8.791	33.6±7.168	10.35±2.450	10±1.633	47.2±9.016	24.1±10.225	63.2±6.812	2.2±0.422		0.8±0.789	1.2±1.033
AF(n=19)
58	F	MS, MI, TI, AI	85	57	39	7	9	72	20	67	3	D, DI	0	1
52	F	MS, MI, TI, AI	58	54	34	11	9	64	64	69	2	DI	2	3
46	F	MS, MI, TI, AS, AI	63	55	38	12	9	85	28	55	3	D, DI, B	0	1
34	F	MS, MI, TI, AI	122	63	38	8	8	37	21	66	2	D, DI, B	0	1
48	F	MS, MI, TI, AI	50	44	27	11	10	60	24	60	3	A, D, DI, C	1	2
54	F	MS, MI, TI, AI	89	40	26	9	10	59	26	54	3	D, DI	0	1
59	M	MS, TI	51	49	34	9	10	52	25	56	3	D, DI	0	0
		AI
55	F	MS, MI, TS, TI, AS, AI	51	32	24	9	10	79	31	60	2	D, DI	0	1
46	F	MS, MI, AI	42	41	29	9	8	61	23	62	3	DI	0	1
62	M	MS, MI, TI, AI	40	48	34	8	7	64	26	79	3	D, DI	2	2
48	M	MI, MS, TI, AI	46	48	33	10	9	60	21	64	2	D, DI	0	0
48	F	MS, MI, TI, AS, AI	57	42	28	11	9	57	20	76	4	DI	0	1
60	F	MI, TI, AI	98	66	34	11	11	86	29	54	2	D, DI, N	0	1
32	M	MI, TI, AI	55	61	39	9	9	56	25	63	3	D, DI	0	0
56	M	MS, MI	82	65	35	10	10	67	23	69	2	D, DI, N	0	0
52	F	MI, TI, AI	51	65	46	13	13	62	21	62	2	D, DI, B	3	4
24	F	MS, MI, TI, AI	60	64	41	9	12	44	21	67	3	D, DI	.00	1.00
60	M	MI, TI, AI	44	63	38	9	9	69	24	69	2	DI	.00	.00
56	F	MS, MI, TI, AS, AI	60	45	30	7	8	70	24	55	3	D, DI	.00	1.00
TOTAL
50±10.301	6M/13F		63.368±21.843^* * P<0.05 vs. SR.	52.737±10.402	34.053±5.690	9.579±1.610	9.474±1.429	63.368±12.321^* * P<0.05 vs. SR.	26.105±9.678	62.526±7.597	2.65±0.671^* * P<0.05 vs. SR.		0.421±0.902	1.105±1.049

	Cytoplasmic DACT1 expression^a a As long as the sample contained cells with DACT1 cytoplasmic staining, the staining was regarded as weak if mean density was <0.025, strong if it was between ≥0.025 and <0.04, and very strong if it was ≥0.04;			P	Nuclear DACT1 expression^b b As long as the sample contained cells with DACT1 nuclear staining, the staining was regarded as positive if the positive cell count was >5% and negative if the mean density was <5%. Chi-square analysis was used to examine the relationships among categorical variables.		P
	Weak	Strong	Very strong	P	Negative	Positive	P
SR	0(0.0%)	3(10.3%)	7(24.1%)	0.037^* * P<0.05.	2(6.9%)	8(27.6%)	0.419
AF	6(20.7%)	2(6.9%)	11(37.9%)		6(20.0%)	13(44.8%)

Indices	Weak	Strong	Very strong	P
EF (%)	63.1667±8.537	60.429±7.138	63.900±7.219	0.618
LA (mm)	58.000±25.856	45.857±10.961	61.400±17.702	0.300
LVD (mm)	54.000±9.737	50.429±11.886	50.700±8.564	0.583
LVS (mm)	35.083±6.037	33.714±8.180	32.600±4.835	0.213
IVS (mm)	10.333±2.146	10.428±1.989	8.850±1.292	0.131
LVPW (mm)	9.917±1.564	9.857±1.574	9.200±1.398	0.508
RA (mm)	57.000±16.321	53.571±14.397	61.700±8.932	0.479
RV (mm)	23.000±3.693	32.429±18.293	23.400±2.119	0.087
Fibrosis degree	0.160±0.109	0.139±0.066	0.164±0.053	0.817

β-catenin expression^b a As long as the sample contained cells with the dishevelled binding antagonist of beta catenin 1 (DACT1) cytoplasmic staining, the staining was regarded as weak if mean density was <0.025, strong if it was between ≥0.025 and <0.04, and very strong if it was ≥0.04.	DACT1 expression^a b As long as the sample contained cells with β-catenin cytoplasmic and/or membrane staining, the total expression level was regarded as weak if mean density was <0.01, strong if it was between ≥0.01 and <0.03, and very strong if it was ≥0.03. The nonparametric Spearman's rank correlation test was used for correlation analysis.			P Spearman's rho
	Weak	Strong	Very strong	P Spearman's rho
Weak	4	1	1	0.028^* * P<0.05. 0.408
Strong	3	2	0
Very strong	5	4	9

Indices	Negative	Very weak	Weak	Strong	Very strong	Spearman's rho P
EF (%)	62.000±8.803	63.928±7.721	58.000±0.000	60.833±6.014	64.000±7.810	0.036 0.855
LA (mm)	58.800±15.643	61.357±24.566	85.000±0.000	43.333±9.004	44.333±6.028	-0.323 0.087
LVD (mm)	50.200±9.757	53.143±10.347	57.000±0.000	55.000±9.166	44.000±9.000	0.015 0.939
LVS (mm)	32.400±3.912	33.928±5.916	39.000±0.000	36.833±6.969	28.667±7.637	0.068 0.724
IVS (mm)	9.400±1.817	10.036±1.623	7.000±0.000	9.667±2.251	11.000±3.000	-0.133 0.491
LVPW (mm)	8.800±0.837	9.857±1.406	9.000±0.000	9.333±2.066	11.000±1.000	0.128 0.508
RA (mm)	70.600±8.849	56.714±14.242	72.000±0.000	51.667±10.367	49.000±9.849	-0.429 0.020^* * P<0.05.
RV (mm)	24.600±2.074	28.714±13.070	20.000±0.000	22.000±3.033	20.000±3.606	-0.378 0.043^* * P<0.05.
Fibrosis degree	0.203±0.051	0.163±0.010	0.208±0.000	0.126±0.054	0.096±0.0149	-0.407 0.028^* * P<0.05.

Cx43 expression^b b As long as the sample contained cells with connexin 43 cytoplasmic and/or membrane staining, the total expression level was regarded as negative if mean density was <0.01, very weak if it was between ≥0.01 and <0.03, weak if it was between ≥0.03 and <0.04, strong if it was between ≥0.04 and <0.05, and very strong if it was between ≥0.05. The nonparametric Spearman's rank correlation test was used for correlation analysis.	DACT1 expression^a a As long as the sample contained cells with the dishevelled binding antagonist of beta catenin 1 (DACT1) cytoplasmic staining, the staining was regarded as weak if mean density was <0.025, strong if it was between ≥0.025 and <0.04, and very strong if it was ≥0.04;		P Spearman's rho
	Weak	Strong/Very strong	P Spearman's rho
Negative	2	3	0.048^* * P<0.05.
Very weak	4	10	0.370
Weak	0	1
Strong	0	6
Very strong	0	3

Authors' roles & responsibilities
JH	The acquisition, analysis, or interpretation of data for the work; final approval of the version to be published
YY	The acquisition, analysis, or interpretation of data for the work; final approval of the version to be published
BH	The acquisition, analysis, or interpretation of data for the work; final approval of the version to be published
GX	Drafting the work or revising it critically for important intellectual content; final approval of the version to be published
RS	Drafting the work or revising it critically for important intellectual content; final approval of the version to be published
LL	Drafting the work or revising it critically for important intellectual content; final approval of the version to be published
JH	The acquisition, analysis, or interpretation of data for the work; final approval of the version to be published
JY	Drafting the work or revising it critically for important intellectual content; final approval of the version to be published
YG	The acquisition, analysis, or interpretation of data for the work; final approval of the version to be published
KW	The acquisition, analysis, or interpretation of data for the work; final approval of the version to be published
ZW	Agreement to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved; final approval of the version to be published

[1] Correspondence Address: Zhongkai Wu, Department of Cardiac Surgery, The First Affiliated Hospital of Sun Yat-Sen University, 58 Zhongshan 2nd Rd., Guangzhou, GD, People’s Republic of China, Zip Code: 510080. E-mail: wuzhk@mail.sysu.edu.cn

Brasil

Brasil

DACT1 Involvement in the Cytoskeletal Arrangement of Cardiomyocytes in Atrial Fibrillation by Regulating Cx43

Abstract

Objective:

Methods:

Results:

Conclusion: