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Rat carotid arteriotomy: c-myc is involved in negative remodelling and apoptosis
Marisa De Feo1, Amalia Forte, Francesco Onorati
1Department of Cardiothoracic Sciences, Second University of Naples, Naples, Italy.
Objective:
c-myc is the main proto-oncogene responsible for restenosis in cardiovascular surgery. The aim of our study was to evaluate the effects of c-myc antisense (AS) phosphorothioate oligodeoxynucleotides (ODNs) in the remodelling process induced by surgical carotid arteriotomy on an experimental rat model.
Methods:
Fifty-five rats with carotid stenosis and apoptosis induced by arteriotomy were submitted to gene expression analysis 4 h after surgery, to TUNEL assay 48 h after surgery and to histological analysis 30 days later.
Results:
AS ODNs induced a 60% decrease in target c-myc mRNA in injured carotid arteries compared to control sense and scrambled ODN-treated carotid arteries (P < 0.05). Histological evaluation revealed that stenosis stimulated by arteriotomy was mainly due to adventitial constrictive remodelling rather than to neointimal hyperplasia, observed only in a limited number of samples. Morphometric analysis showed that lumen area in c-myc AS ODN-treated carotid arteries was 35% greater than in control arteries (P < 0.05), whereas the media/lumen area ratio showed a 63% reduction in AS ODN-treated carotid arteries in comparison to control arteries (P < 0.05). Surgical injury affected the expression of apoptosis-related genes Bcl-2, Bax, Bcl-xL and Bcl-xS, inducing a mean 3.5-fold decrease in the Bcl-2/ Bax ratio and a 9-fold decrease in the Bcl-xL/S ratio 4 h after injury as compared with uninjured carotid arteries. TUNEL assay experiments revealed increased apoptosis in AS ODN-treated carotid arteries in comparison to control carotid arteries.
Conclusions:
c-myc AS ODNs reduce the negative remodelling induced by arteriotomy. The imbalance between proliferative stimulus represented by surgery and the c-myc mRNA decrease induced greater apoptosis in AS ODN-treated carotid arteries without further affecting mRNA levels of Bcl-2, Bax, Bcl-xL and Bcl-xS genes.
Insights
C-myc antisense oligodeoxynucleotides (AS ODNs) effectively reduced c-myc mRNA in carotid arteries after surgery. This targeted gene therapy improved lumen area and reduced arterial remodeling, promoting healing.
Area of Science:
- Cardiovascular Biology
- Molecular Medicine
- Gene Therapy
Background:
- C-myc is a key proto-oncogene implicated in restenosis following cardiovascular surgery.
- Arteriotomy-induced carotid stenosis involves complex remodeling processes.
- Understanding c-myc's role is crucial for developing therapeutic strategies.
Purpose of the Study:
- To investigate the therapeutic potential of c-myc antisense (AS) phosphorothioate oligodeoxynucleotides (ODNs).
- To evaluate the impact of AS ODNs on arterial remodeling after surgical carotid arteriotomy in a rat model.
- To assess the effects on c-myc mRNA levels, apoptosis, and histological changes.
Main Methods:
- Gene expression analysis of c-myc mRNA in injured carotid arteries.
- TUNEL assay to quantify apoptosis at 48 hours post-surgery.
- Histological and morphometric analysis of arterial structure at 30 days.
Main Results:
- AS ODNs significantly decreased c-myc mRNA by 60% in injured arteries.
- Arterial stenosis was primarily due to adventitial constrictive remodeling.
- AS ODN treatment increased lumen area by 35% and reduced media/lumen ratio by 63%.
Conclusions:
- C-myc AS ODNs effectively mitigate negative arterial remodeling induced by arteriotomy.
- The reduction in c-myc mRNA promoted apoptosis, counteracting proliferative stimuli.
- AS ODNs offer a promising therapeutic approach for preventing post-surgical restenosis.
