Heat shock provides delayed protection against oxidative injury in cultured human umbilical vein endothelial cells.
During both mild and severe ischemia, vascular endothelial cells lining large and small vessels of the ischemic organ are exposed to oxygen-derived free radicals resulting in oxidative damage to the organ. Heat shock has been shown to induce thermotolerance and also protect against ischemic injury, possibly via increased synthesis of heat shock proteins (HSPs). We hypothesized that heat shock preconditioning may protect human endothelial cells against oxidative damage. Cultured human umbilical vein endothelial cells (HUVEC) were subjected to heat shock (42 degrees C, 1 h) and allowed to recover for 2 or 20 h, at which times the cells were oxidatively stressed for 1 h by exposing them to 100-200 mumol/l of hydrogen peroxide (H2O2). Cellular damage was assessed immediately and 18 h later by morphology and release of lactate dehydrogenase (LDH). No protection of HUVEC was seen using the 2-hour recovery interval, but a significant protection (P < 0.05) was observed after the 20-hour delay. Northern blot analysis at 1 and 2 h after heating showed induction of HSP-70 mRNA. Western blot analysis demonstrated a significant increase in HSP-72 protein after 2 as well as 20 h of recovery from heat shock, although the amounts of protein at the two times were not significantly different. Furthermore, no differences in the activity of the antioxidant enzyme catalase were observed between heated and unheated HUVEC at 2 and 20 h after heat preconditioning. Thus, heat shock preconditioning induces delayed protection against oxidative injury in HUVEC, and the mechanism of protection appears to involve more than the expression of HSP-72 or activity of catalase.
Gill
R R
RR
Department of Physiology, Medical College of Virginia, Virginia Commonwealth University, Richmond 23298, USA.
Gbur
C J
CJ
Fisher
B J
BJ
Hess
M L
ML
Fowler
A A
AA
Kukreja
R C
RC
Sholley
M M
MM
D015152:Blotting, Northern; D015153:Blotting, Western; D002374:Catalase / Q000502:physiology; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000502:physiology*; D050884:HSP72 Heat-Shock Proteins; D006360:Heat-Shock Proteins / Q000494:pharmacology; D018869:Heat-Shock Response / Q000502:physiology*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D018384:Oxidative Stress / Q000502:physiology*; D013696:Temperature; D013997:Time Factors; D014471:Umbilical Veins / Q000502:physiology*