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J Appl Physiol (April 16, 2004). doi:10.1152/japplphysiol.00194.2004
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Submitted on February 25, 2004
Accepted on April 10, 2004

Geldanamycin treatment inhibits hemorrhage-induced increases in KLF6 and iNOS expression in un-resuscitated mouse organs: Role of inducible HSP-70

Juliann G Kiang1*, Phillip D Bowman2, Brian W Wu3, Nyasa Hampton3, Andrew G Kiang3, Baiteng Zhao2, Yuang-Taung Juang4, James L Atkins3, and George C Tsokos4

1 Division of Military Casualty Research, Cellular Injury, Walter Reed Army Institute of Research, Silver Spring, MD, USA; Department of Medicine, Uniformed Services University of The Health Sciences, Bethesda, MD, USA; Department of Pharmacology, Uniformed Services University of The Health Sciences, Bethesda, MD, USA
2 US Army Institute of Surgical Research, San Antonia, TX, USA
3 Division of Military Casualty Research, Cellular Injury, Walter Reed Army Institute of Research, Silver Spring, MD, USA
4 Division of Military Casualty Research, Cellular Injury, Walter Reed Army Institute of Research, Silver Spring, MD, USA; Department of Medicine, Uniformed Services University of The Health Sciences, Bethesda, MD, USA

* To whom correspondence should be addressed. E-mail: Juliann.Kiang{at}na.amedd.army.mil.

The aim of this study was to determine if hemorrhage affected the levels of a variety of stress proteins and if changes could be inhibited by drugs reported to provide protection from ischemia/reperfusion injury. Male Swiss Webster mice were subjected to a 40% hemorrhage without resuscitation. Caspase-3 activity increased within 1 h after hemorrhage and reached a maximum after 6 h in lung, jejunum, heart, and kidney, but not in the liver or brain. Western blot analysis indicated that c-JUN (an AP-1 protein), Kruppel-like factor 6 transcription factor (KFL6), and inducible nitric oxide synthase (iNOS) were upregulated sequentially, in that order. Pretreatment of mice with geldanamycin (GA, i.p.) 16 h before hemorrhage significantly inhibited the hemorrhage-induced increase in caspase-3 activity in lung, jejunum, kidney, and heart, while caffeic acid phenethyl ester (CAPE, i.p.) was not effective at reducing caspase-3 activity. Pretreatment with GA also effectively inhibited the expression of the proteins KLF6 and iNOS, while CAPE did not. GA pretreatment increased inducible heat shock protein 70 kDa (HSP-70i) but not HSP 90 kDa in both sham and hemorrhagic tissues. The overexpressed HSP-70i formed complexes with KLF6 and iNOS. These results suggest that GA may be therapeutically useful for reducing hemorrhage-induced injury when used as a pre-surgical treatment or when added to resuscitation fluids.




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