Reduced hexokinase II impairs muscle function 2 wk after ischemia-reperfusion through increased cell necrosis and fibrosis

• Published in: Journal of applied physiology (1985) 2012-08, Vol.113 (4), p.608-618
• Main Authors: Smeele, Kirsten M, Eerbeek, Otto, Schaart, Gert, Koeman, Anneke, Bezemer, Rick, Nelson, Jessica K, Ince, Can, Nederlof, Rianne, Boek, Maxim, Laakso, Markku, de Haan, Arnold, Drost, Maarten R, Hollmann, Markus W, Zuurbier, Coert J
• Format: Article
• Language: English
• Subjects: Animals; Apoptosis; bcl-2-Associated X Protein - metabolism; Disease Models, Animal; Down-Regulation; Enzymes; Fibrosis; Gangrene; Glycogen - metabolism; Hexokinase - deficiency; Hexokinase - genetics; Hindlimb; Ischemia; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Microcirculation; Mitochondria, Muscle - metabolism; Mitochondria, Muscle - pathology; Muscle Fatigue; Muscle Strength; Muscle, Skeletal - blood supply; Muscle, Skeletal - enzymology; Muscle, Skeletal - pathology; Muscle, Skeletal - physiopathology; Musculoskeletal system; Necrosis; Neovascularization, Physiologic; Oxidative Stress; Pathology; Recovery of Function; Regeneration; Regional Blood Flow; Reperfusion Injury - enzymology; Reperfusion Injury - genetics; Reperfusion Injury - pathology; Reperfusion Injury - physiopathology; Rodents; Time Factors
• ISSN: 8750-7587; 1522-1601
• DOI: 10.1152/japplphysiol.01494.2011

We previously demonstrated that hexokinase (HK) II plays a key role in the pathophysiology of ischemia-reperfusion (I/R) injury of the heart (Smeele et al. Circ Res 108: 1165-1169, 2011; Wu et al. Circ Res 108: 60-69, 2011). However, it is unknown whether HKII also plays a key role in I/R injury and healing thereafter in skeletal muscle, and if so, through which mechanisms. We used male wild-type (WT) and heterozygous HKII knockout mice (HKII(+/-)) and performed in vivo unilateral skeletal muscle I/R, executed by 90 min hindlimb occlusion using orthodontic rubber bands followed by 1 h, 1 day, or 14 days reperfusion. The contralateral (CON) limb was used as internal control. No difference was observed in muscle glycogen turnover between genotypes at 1 h reperfusion. At 1 day reperfusion, the model resulted in 36% initial cell necrosis in WT gastrocnemius medialis (GM) muscle that was doubled (76% cell necrosis) in the HKII(+/-) mice. I/R-induced apoptosis (29%) was similar between genotypes. HKII reduction eliminated I/R-induced mitochondrial Bax translocation and oxidative stress at 1 day reperfusion. At 14 days recovery, the tetanic force deficit of the reperfused GM (relative to control GM) was 35% for WT, which was doubled (70%) in HKII(+/-) mice, mirroring the initial damage observed for these muscles. I/R increased muscle fatigue resistance equally in GM of both genotypes. The number of regenerating fibers in WT muscle (17%) was also approximately doubled in HKII(+/-) I/R muscle (44%), thus again mirroring the increased cell death in HKII(+/-) mice at day 1 and suggesting that HKII does not significantly affect muscle regeneration capacity. Reduced HKII was also associated with doubling of I/R-induced fibrosis. In conclusion, reduced muscle HKII protein content results in impaired muscle functionality during recovery from I/R. The impaired recovery seems to be mainly a result of a greater susceptibility of HKII(+/-) mice to the initial I/R-induced necrosis (not apoptosis), and not a HKII-related deficiency in muscle regeneration.