Negative inotropic effects of high-mobility group box 1 protein in isolated contracting cardiac myocytes.
High-mobility group box 1 (HMGB1) released from necrotic cells or macrophages functions as a late inflammatory mediator and has been shown to induce cardiovascular collapse during sepsis. Thus far, however, the effect(s) of HMGB1 in the heart are not known. We determined the effects of HMGB1 on isolated feline cardiac myocytes by measuring sarcomere shortening in contracting cardiac myocytes, intracellular Ca2+ transients by using fluo-3, and L-type calcium currents by using whole cell perforate configuration of the patch-clamp technique. Treatment of isolated myocytes with HMGB1 (100 ng/ml) resulted in a 70% decrease in sarcomere shortening and a 50% decrease in the height of the peak Ca2+ transient within 5 min (P < 0.01). The immediate negative inotropic effects of HMGB1 on cell contractility and calcium homeostasis were partially reversible upon washout of HMGB1. A significant inhibition of the inward l-type calcium currents was also documented by the patch-clamp technique. HMGB1 induced the PKC-epsilon translocation, and a PKC inhibitor significantly attenuated the negative inotropic effects of HMGB1. These studies show for the first time that HMGB1 impairs sarcomere shortening by decreasing calcium availability in cardiac myocytes through modulating membrane calcium influx and suggest that HMGB1 maybe acts as a novel myocardial depressant factor during cardiac injury.
Tzeng
Huei-Ping
HP
Department of medicine, Winters Center for Heart Failure Research, Houston, TX 77030, USA.
Fan
Jinping
J
Vallejo
Jesus G
JG
Dong
Jian Wen
JW
Chen
Xiongwen
X
Houser
Steven R
SR
Mann
Douglas L
DL
D000814:Aniline Compounds; D000818:Animals; D020746:Calcium Channels, L-Type / Q000187:drug effects; D002415:Cats; D002469:Cell Separation; D016207:Cytokines / Q000502:physiology; D003864:Depression, Chemical; D005456:Fluorescent Dyes; D024243:HMGB1 Protein / Q000494:pharmacology*; D009200:Myocardial Contraction / Q000187:drug effects*; D032383:Myocytes, Cardiac / Q000187:drug effects*; D018408:Patch-Clamp Techniques; D051744:Protein Kinase C-epsilon / Q000378:metabolism; D021381:Protein Transport / Q000502:physiology; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic / Q000037:antagonists & inhibitors / Q000276:immunology; D012518:Sarcomeres / Q000187:drug effects; D051197:Toll-Like Receptor 4 / Q000037:antagonists & inhibitors / Q000276:immunology; D014966:Xanthenes
D000814:Aniline Compounds; D020746:Calcium Channels, L-Type; D016207:Cytokines; D005456:Fluorescent Dyes; D024243:HMGB1 Protein; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic; D051197:Toll-Like Receptor 4; D014966:Xanthenes; C059715:Fluo-3; D051744:Protein Kinase C-epsilon