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Adenoviral transfer of PKGIα; attenuates apoptosis and necrosis in adipose derived stem cells
Author(s) -
Hoke Nicholas,
Salloum Fadi N,
Das Anindita,
Chau Vinh Q,
Wehman Paul B,
Kukreja Rakesh C
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.lb34
Subject(s) - apoptosis , tunel assay , myocyte , necrosis , ischemia , medicine , stem cell , microbiology and biotechnology , chemistry , biology , biochemistry
Adenoviral transfer of protein kinase G‐Iα (PKGIα) protects myocytes from ischemia but the effect of PKGIα on adipose derived stem cells (ASCs) remains unknown. We hypothesize that PKGIα will attenuate necrosis and apoptosis of ASCs in an ischemic environment. ASCs were infected with adenoviral vectors containing PKGIα or a mutant then subjected to simulated ischemia (SI) and reoxygenation (RO) for necrosis or apoptosis studies. To determine the role of PKG overexpression in vivo , adult male ICR mice underwent permanent left coronary artery occlusion before receiving treatment of 2x10^5 cells overexpressing PKGIα or mutant. 2 weeks after surgery, transthoracic echocardiography (TTE) was performed to assess left ventricular (LV) function. PKGIα overexpression reduced necrosis as seen by a decrease in trypan blue positive cells and LDH release when compared to SI/RO control ( Fig 1 A, B). TUNEL revealed that apoptosis was also significantly reduced in cells overexpressing PKGIα (5.1± 0.47%) as compared to SI/RO control (12.8±1.07%). ASCs infected with the inactive PKGIαK390A mutant lacked protection. TTE results showed reduced LV dilatation and preservation of ejection fraction with PKG overexpression ( Fig 1 C, D). Therefore, adenoviral gene transfer of PKGIα may provide a novel strategy for protecting ASCs against ischemia which could translate to a favorable function in hearts following ischemia. 1