Quantification of DNA Damage in Different Tissues in Rats with Heart Failure

Stefani, Giuseppe Potrick; Nunes, Ramiro Barcos; Rossato, Douglas Dalcin; Hentschke, Vitor Scotta; Domenico, Marlise Di; Lago, Pedro Dal; Rhoden, Cláudia Ramos

doi:10.36660/abc.20180198

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Original Article • Arq. Bras. Cardiol. 114 (2) • Feb 2020 • https://doi.org/10.36660/abc.20180198 copy

Quantification of DNA Damage in Different Tissues in Rats with Heart Failure

Authorship SCIMAGO INSTITUTIONS RANKINGS

Abstract

Background:

Chronic heart failure (CHF) is a complex syndrome which comprises structural and functional alterations in the heart in maintaining the adequate blood demand to all tissues. Few investigations sought to evaluate oxidative DNA damage in CHF.

Objective:

To quantify the DNA damage using the comet assay in left ventricle (LV), lungs, diaphragm, gastrocnemius and soleus in rats with CHF.

Methods:

Twelve male Wistar rats (300 to 330 g) were selected for the study: Sham (n = 6) and CHF (n = 6). The animals underwent myocardial infarction by the ligation of the left coronary artery. After six weeks, the animals were euthanized. It was performed a cell suspension of the tissues. The comet assay was performed to evaluate single and double strand breaks in DNA. Significance level (p) considered < 0.05.

Results:

The CHF group showed higher values of left ventricle end-diastolic pressure (LVEDP), pulmonary congestion, cardiac hypertrophy and lower values of maximal positive and negative derivatives of LV pressure, LV systolic pressure (p < 0.05). CHF group showed higher DNA damage (% tail DNA, tail moment and Olive tail moment) compared to Sham (p < 0.001). The tissue with the highest damage was the soleus, compared to LV and gastrocnemius in CHF group (p < 0.05).

Conclusion:

Our results indicates that the CHF affects all tissues, both centrally and peripherically, being more affected in skeletal muscle (soleus) and is positively correlated with LV dysfunction.

Keywords:
Heart Failure; Rats; Rats Inbred Strains; Tissue Distribution; DNA Damage; Comet Assay

Variables	Sham	CHF
Initial Body Mass (g)	330.25 ± 17.24	328.29 ± 18.12
Final Body Mass (g)	400.50 ± 29.61	356.38 ± 32.23
Infarcted Area (%)	---	36.39 ± 8.11
MM/BM (mg/g)	2.56 ± 0.08	3.29 ± 0.46^* * p < 0.05 compared to the Sham group.
LV/BM (mg/g)	1.88 ± 0.21	2.36 ± 0.47^* * p < 0.05 compared to the Sham group.
RV/BM (mg/g)	0.58 ± 0.12	1.40 ± 1.01^* * p < 0.05 compared to the Sham group.
Pulmonary Congestion (%)	65.67 ± 9.34	87.31 ± 3.36^* * p < 0.05 compared to the Sham group.
Hepatic Congestion (%)	70.46 ± 1.05	71.43 ± 1.07

Variables	Sham	CHF
MBP (mmHg)	93.01 ± 14.70	76.78 ± 5.83^* * p < 0.05 compared to the Sham group.
DBP (mmHg)	73.54 ± 16.28	67.54 ± 7.15
SBP (mmHg)	99.75 ± 20.91	85.93 ± 5.51
Heart Rate (bpm)	253.56 ± 70.84	245.19 ± 57.69
LVEDP (mmHg)	5.40 ± 2.26	32.55 ± 5.32^* * p < 0.05 compared to the Sham group.
LVSP (mmHg)	104.24 ± 6.03	89.15 ± 3.15^* * p < 0.05 compared to the Sham group.
+ dP/dt_max (mmHg/s)	6,264.33 ± 1,566.47	4,281.63 ± 708.75^* * p < 0.05 compared to the Sham group.
- dP/dt_max (mmHg/s)	5,209.63 ± 1,274.09	2,823.80 ± 540.65^* * p < 0.05 compared to the Sham group.

		Sham			CHF
	% Tail DNA	Tail Moment	Olive Tail Moment	% Tail DNA	Tail Moment	Olive Tail Moment
Left Ventricle	7.65 ± 3.35	0.77 ± 0.44	1.37 ± 0.59	33.29 ± 7.70^* * = p < 0.01 versus Sham in relation to the variable and its corresponding tissue.	10.51 ± 3.31^* * = p < 0.01 versus Sham in relation to the variable and its corresponding tissue.	7.04 ± 1.71^* * = p < 0.01 versus Sham in relation to the variable and its corresponding tissue.
Lungs	17.86 ± 3.93	6.76 ± 2.59	7.31 ± 2.15	36.20 ± 5.17^* * = p < 0.01 versus Sham in relation to the variable and its corresponding tissue.	23.30 ± 7.25^* * = p < 0.01 versus Sham in relation to the variable and its corresponding tissue.	19.10 ± 4.65^* * = p < 0.01 versus Sham in relation to the variable and its corresponding tissue.
Diaphragm	6.86 ± 2.63	1.40 ± 0.93	1.82 ± 0.79	41.23 ± 13.86^* * = p < 0.01 versus Sham in relation to the variable and its corresponding tissue.	14.06 ± 6.51^* * = p < 0.01 versus Sham in relation to the variable and its corresponding tissue.	9.82 ± 3.03^* * = p < 0.01 versus Sham in relation to the variable and its corresponding tissue.
Gastrocnemius	7.63 ± 4.66	1.04 ± 0.88	1.43 ± 0.70	28.07 ± 15.53^* * = p < 0.01 versus Sham in relation to the variable and its corresponding tissue.	8.69 ± 5.14^* * = p < 0.01 versus Sham in relation to the variable and its corresponding tissue.	6.17 ± 3.53^* * = p < 0.01 versus Sham in relation to the variable and its corresponding tissue.
Soleus	11.54 ± 2.46	1.53 ± 0.96	1.84 ± 0.76	55.79 ± 11.53^* * = p < 0.01 versus Sham in relation to the variable and its corresponding tissue.	20.90 ± 5.32^* * = p < 0.01 versus Sham in relation to the variable and its corresponding tissue.	12.83 ± 3.68^* * = p < 0.01 versus Sham in relation to the variable and its corresponding tissue.

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[1] Mailing Address: Giuseppe Potrick Stefani, Universidade Federal de Ciências da Saúde de Porto Alegre - Sarmento Leite street, 245. Postal Code 90050-170, Porto Alegre, RS - Brazil. E-mail: gpotrick@gmail.com