Connexin43 and Akt expression improves skeletal myoblast viability under conditions of hyperthermic stress

• Published in: Cardiology
• Main Authors: Antanavičiūtė, Ieva, Žūkienė, Rasa, Buzaitė, Odeta, Kalvelytė, Audronė, Mildažienė, Vida, Skeberdis, Vytenis Arvydas
• Format: Conference Object
• Language: English
• Published: 2009
• Subjects: Connexin 43; Proto-oncogene proteins c-akt; Myoblasts; skeletal; Fever; Metabolism; Tezės Clarivate Analytics Web of Science / Theses in Clarivate Analytics Web of Science (T1a); Biologija / Biology (N010); Iceland
• Online Access: http://content.karger.com/ProdukteDB/produkte.asp?Aktion=ShowPDF&ArtikelNr=223370&Ausgabe=249596&ProduktNr=223832&filename=223370.pdf; https://doi.org/10.1159/000223370

Abstracts and program XXII Nordic-Baltic congress of cardiology : Reykjavik, Iceland, June 3-5, 2009 / Guest editors : David O. Arnar, Karl Andersen We use autologic skeletal myoblasts for regeneration of cardiac muscle after experimentally induced infarction in rabbit heart. To improve electrical integration and antiapoptotic properties of myoblasts, and to prevent the incidence of ventricular tachyarrhythmias we transfected the myoblasts with connexin43 (Cx43) or protein kinase B (Akt) genes. Since the transplantation of the cells is often followed by inflammation, we examined the resistance of control and Cx43 or Akt transfected cells to hyperthermic shock. We demonstrated that measurement of NAD(P)H fluorescence may be used for evaluation of biochemical status and viability of cellular population. The effect of hyperthermia on amounts of adenine and pyridine nucleotides was determined in cell extracts by Alliance HPLC system (Waters Corporation). To examine the effect of hyperthermia on cell viability of control and Cx43 or Akt expressing myoblasts, standard Trypan blue test was performed in parallel with measurements of NAD(P)H fluorescence (fluorimeter Tecan GENios ProTM). Viability of Cx43-transfected cells was about 10% higher under different hyperthermic incubations (40–50ºC) comparing with control (not transfected) cells. Correlation coefficient between myoblast viability and NAD(P)H fluorescence intensity in 4 different cell lines in average was 0,84. Transfection with Akt also improved myoblast viability under the same hyperthermic conditions (by 20–26%). Chromatographic analysis confirmed that NAD(P)H fluorescence decays in parallel with decrease of amounts of the reduced forms of pyridine nucleotides under conditions of hyperthermic stress. We conclude that transfection with connexin43 and Akt genes may increase the resistance of myoblast cell lines to hyperthermic stress Biochemijos institutas Biochemijos katedra Biologijos katedra Kauno medicinos universiteto Kardiologijos institutas Vytauto Didžiojo universitetas