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01 December 2011

Heat shock attenuates VEGF expression in three-dimensional myoblast sheets deteriorating therapeutic efficacy in heart failure

Mona AugustinABCDEF, Muhammad Ali Asim MaharABCDEF, Päivi LakkistoCD, Ilkka TikkanenG, Antti VentoEF, Tommi PatilaCDEF, Ari HarjulaG

DOI: 10.12659/MSM.882120

Med Sci Monit 2011; 17(12): BR345-353

Abstract

Background: Myoblast sheet transplantation is a promising novel treatment for ischemic heart failure. The aim of this study was to test the hypothesis that heat shock (HS) pre-treatment affects the angiogenic properties of myoblast sheets in vivo and in vitro.
Material/Methods: We studied HS preconditioning of L6 myoblast sheets in relation to their apoptosis, proliferation, and vascular endothelial growth factor (VEGF)-associated responses under normoxia and under hypoxia in vitro. In vivo evaluation of their therapeutic effect was performed with 60 male Wistar rats divided into 3 groups (20 each): sole left anterior descending (LAD) ligation (control); LAD ligation and non-conditioned sheet transplantation (L6 No-Shock); and LAD ligation and L6-heat shock conditioned sheet transplantation (L6 Heat-Shock). Left ventricular function was evaluated by echocardiography after 3, 10, and 28 days.
Results: Expression of HSP70/72 was strongly induced 24 hours after HS, and thereafter it decreased notably during 72 hours in hypoxia. Under normal growth conditions, HSP70/72 expression remained stable. HS delayed apoptosis-associated caspase-3 expression during 24-hour hypoxia compared to non-treated controls. However, VEGF expression reduced significantly in the heat shock pretreated sheets. Ejection fraction of the L6-myoblast HS pre-treatment group (L6 Heat-Shock) decreased gradually during follow-up, in the same pattern as the controls. However, these functional parameters improved in the L6-myoblast normal sheet group (L6 No-Shock) at the tenth day and remained significantly better.
Conclusions: HS protects myoblast sheets from hypoxia-associated apoptosis in vitro, but reduces VEGF expression of the sheet, leading to lower therapeutic effect in heart failure.

Keywords: Oxidative Stress, Myocardial Infarction - ultrasonography, Myoblasts - transplantation, Heat-Shock Response, Heart Failure - ultrasonography, HSP72 Heat-Shock Proteins - metabolism, HSP70 Heat-Shock Proteins - metabolism, Cell Line, Cell Hypoxia, Cell Culture Techniques - methods, Caspase 3 - metabolism, Animals, Proliferating Cell Nuclear Antigen - metabolism, Rats, Rats, Wistar, Stroke Volume, Vascular Endothelial Growth Factor A - secretion

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750