This Article Abstract Full Text (PDF) Data Supplement All Versions of this Article: 110/23/3560    most recent 01.CIR.0000143082.63063.33v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Chan, J. Y.H. Articles by Chan, S. H.H. Search for Related Content PubMed PubMed Citation Articles by Chan, J. Y.H. Articles by Chan, S. H.H. Pubmed/NCBI databases Compound via MeSH Substance via MeSH Related Collections Animal models of human disease Cell signalling/signal transduction

(Circulation. 2004;110:3560-3566.)
© 2004 American Heart Association, Inc.

Molecular Cardiology

Heat Shock Protein 70 Confers Cardiovascular Protection During Endotoxemia via Inhibition of Nuclear Factor-B Activation and Inducible Nitric Oxide Synthase Expression in the Rostral Ventrolateral Medulla

From the Department of Medical Education and Research (J.Y.H.C., C.C.O., L.L.W.), Kaohsiung Veterans General Hospital, Kaohsiung, and Center for Neuroscience (S.H.H.C.), National Sun Yat-sen University, Kaohsiung, Taiwan, Republic of China.

Correspondence to Samuel H.H. Chan, PhD, Center for Neuroscience, National Sun Yat-sen University, Kaohsiung 80424, Taiwan, Republic of China. E-mail schan{at}mail.nsysu.edu.tw

Received May 15, 2004; revision received July 22, 2004; accepted July 27, 2004.

	Abstract

Top Abstract Introduction Methods Results Discussion References

 
Background— Overproduction of nitric oxide (NO) by inducible NO synthase (iNOS) in the rostral ventrolateral medulla (RVLM), where sympathetic premotor neurons are located, plays a pivotal role in the manifestation of fatal cardiovascular depression during endotoxemia. The iNOS gene is regulated transcriptionally by nuclear factor-B (NF-B) activation. The present study tested the hypothesis that heat shock protein 70 (HSP70) may confer protection against sepsis-induced circulatory fatality via inhibition of iNOS gene expression in the RVLM through prevention of NF-B activation.

Methods and Results— Adult male Sprague-Dawley rats subjected to a brief hyperthermic heat shock (42°C for 15 minutes) exhibited significant upregulation of HSP70 in the RVLM. Brief heat shock preconditioning also significantly suppressed iNOS mRNA or protein surge and alleviated hypotension, bradycardia, and reduction in neurogenic sympathetic vasomotor activity manifested during experimental endotoxemia induced by intravenous administration of Escherichia coli lipopolysaccharide. An increase in DNA binding activity and nuclear translocation of transcription factor NF-B were detected during endotoxemia. Heat shock preconditioning significantly decreased DNA binding activity of NF-B, which was reversed by microinjection of an hsp70 antisense oligonucleotide bilaterally into the RVLM. Heat shock preconditioning also blocked inhibitory B (IB) kinase activity or degradation of IB in the RVLM during endotoxemia.

Conclusions— We conclude that HSP70 confers protection against sepsis-related circulatory fatality via inhibition of iNOS gene expression in the RVLM through prevention of NF-B activation in cellular processes that include prevention of IB kinase activation and inhibition of IB degradation.

Key Words: blood pressure • cardiovascular diseases • nitric oxide synthase • shock • signal transduction

	Introduction

Top Abstract Introduction Methods Results Discussion References

 
Cardiovascular depression during sepsis remains a significant cause of morbidity and mortality.¹ One well-known mediator of sepsis-induced circulatory failure at the peripheral vasculature is nitric oxide (NO) produced via activation of inducible NO synthase (iNOS) in macrophages.^2,3 In the central nervous system, we demonstrated recently that by eliciting a reduction in sympathetic vasomotor outflow and arterial pressure,^4,5 overproduction of NO by iNOS⁵ and formation of peroxynitrite by reacting NO with superoxide anion⁶ in the rostral ventrolateral medulla (RVLM), where sympathetic premotor neurons are located,⁷ play a pivotal role in the fatal cardiovascular depression associated with endotoxemia.

The iNOS gene is regulated transcriptionally in part by nuclear factor-B (NF-B) activation.⁸ NF-B is sequestered in the cytoplasm in an inactive state because of its association with inhibitory B (IB).⁹ Under stress conditions,^10,11 phosphorylation of IB by IB kinase (IKK) occurs, which leads to degradation of IB and disruption of the NF-B/IB complex. The dissociated NF-B subsequently translocates from the cytoplasm to the nucleus, where this transcription factor binds to the B promoter region of target genes, including iNOS.^8,12 Accordingly, prevention of NF-B activation inhibits iNOS expression.^13–15

Heat shock (HS) response consists of expression of a family of highly conserved proteins known as heat shock proteins (HSPs).¹⁶ Thermal preconditioning induces expression of HSP70 and reduces iNOS expression elicited by bacterial endotoxin in various cell types.^15,17,18 HS inhibits iNOS gene expression by transcriptional mechanisms that involve the NF-B/IB pathway.^15,19,20 It follows that HSP70 induced by HS may confer protection against sepsis-related circulatory depression via inhibition of iNOS gene expression in the RVLM through prevention of NF-B activation. This hypothesis was validated in the present study. We further demonstrated that HSP70-induced cardiovascular protection is related to stabilization of IB, possibly through prevention of IKK activation and inhibition of IB degradation.

	Methods

Top Abstract Introduction Methods Results Discussion References

 
A brief summary of the experimental strategy is provided. Detailed information on materials and methods can be found in the online Data Supplement.

HSP70 Expression in Ventrolateral Medulla After HS
Adult Sprague-Dawley rats anesthetized with pentobarbital (50 mg/kg) were subjected to hyperthermic HS by maintenance of the core temperature of animals at 42±0.5°C for 15 minutes.^21,22 The ventrolateral medulla that contained bilateral RVLM was removed, and Western blot analysis was performed to detect HSP70 expression at various time intervals after HS.

Effect of HS on Cardiovascular Responses During Experimental Endotoxemia
The effect of HS preconditioning on temporal changes in mean systemic arterial pressure (MSAP), heart rate (HR), or power density of vasomotor components of systemic arterial pressure (SAP) signals after intravenous administration of lipopolysaccharide (LPS; 20 mg/kg, serotype 0111:B4; Sigma) was routinely monitored for 6 hours in animals maintained under propofol (30 mg · kg^–1 · h^–1) anesthesia.²³ We confirmed the presence of a causative relationship between HS-induced HSP70 expression and cardiovascular protection during experimental endotoxemia by bilateral microinjection of an antisense oligonucleotide that targets against the coding region (nt 61 to 78) of hsp70 gene into the RVLM immediately after HS.²²

Effect of HS on iNOS mRNA or Protein Expression in Ventrolateral Medulla During Experimental Endotoxemia
Expression of iNOS mRNA or protein in the ventrolateral medulla from LPS-treated animals that received HS before endotoxemia was measured by reverse transcription-polymerase chain reaction or Western blot analysis.⁶

Activation of NF-B in Ventrolateral Medulla During Experimental Endotoxemia and Its Modulation by HS
We measured NF-B DNA binding activity in nuclear protein from ventrolateral medulla during endotoxemia and its modulation by HS using electrophoresis mobility shift assay.²⁴ Antiserum against NF-B p65, p50, or c-Rel subunit was used in supershift assay to study translocation of NF-B subunits to the nucleus. We again confirmed the presence of a causative relationship between HS-induced HSP70 expression and nuclear translocation of NF-B by antisense hsp70 oligonucleotide treatment. We also evaluated the significance of the activated NF-B in cardiovascular responses and iNOS expression during endotoxemia by blocking the B element in the nucleus with bilateral microinjection of the double-stranded B decoy DNA into the RVLM.²⁴

Mechanisms Underlying Prevention of NF-B Nuclear Translocation by HS
Two approaches were used to delineate the mechanisms that underlie HS-induced prevention of NF-B nuclear translocation during endotoxemia. An immune complex kinase assay was used to determine the effects of HS on the activity of IKK activity, which phosphorylates IB, leading to dissociation of NF-B from NF-B/IB complex.²⁴ Western blot analysis was performed to detect the degradation and reexpression of IB in the ventrolateral medulla after phosphorylation by IKK.

	Results

Top Abstract Introduction Methods Results Discussion References

 
Hyperthermic HS Induced Temporal Changes in HSP70 Expression in the Ventrolateral Medulla That Were Reversed by Antisense hsp70 Oligonucleotide Treatment
Western blot analysis (Figure 1A) revealed that HSP70 in the ventrolateral medulla underwent a significant increase at 8 hours, followed by a progressive augmentation that peaked at 24 hours, and a return to baseline by 48 hours after exposure of animals to a brief HS. In normothermic (NT) controls, in which animals were similarly anesthetized but without subsequent hyperthermic treatment, HSP70 expression was comparable to that of sham controls at all time intervals examined (data not shown). Bilateral microinjection of an antisense hsp70 oligonucleotide (50 pmol) into the RVLM significantly attenuated the upregulation of HSP70 expression in the ventrolateral medulla 16 or 24 hours after HS (Figure 1B). Treatment with sense (50 pmol) or scrambled (50 pmol) hsp70 oligonucleotide was ineffective.

View larger version (29K): [in this window] [in a new window]   Figure 1. A, Representative Western blots of HSP70 (inset) or HSP70 levels detected from ventrolateral medulla in rats subjected to hyperthermic HS or in NT controls. Note that for clarity, only results on samples obtained 24 hours after NT were presented, although samples collected at all corresponding time intervals exhibited levels of HSP70 that were comparable to sham controls (C). B, Representative Western blots of HSP70 (inset) and HSP70 levels detected from ventrolateral medulla 16 or 24 hours after animals were subjected to HS alone or with additional bilateral microinjection of antisense (ASON), sense (SON), or scrambled (SRON) hsp70 oligonucleotide (50 pmol) into RVLM. Note that samples taken 16 hours after HS from SON- or SRON-treated groups are not shown because they essentially duplicated those at 24 hours. Values are mean±SEM of quadruplicate analyses on samples pooled from 4 to 5 animals in each group. *P<0.05 vs C (A) or NT (B) group, #P<0.05 vs corresponding HS group in Scheffé multiple range analysis.

Hyperthermic HS Alleviated Cardiovascular Depression During Experimental Endotoxemia and Its Antagonism by Antisense hsp70 Oligonucleotide Treatment
Similar to our previous findings,⁶ systemic administration of LPS (20 mg/kg) characteristically promoted hypotension, bradycardia, and a reduction in the power density of the vasomotor component of the SAP spectrum, our experimental index for neurogenic sympathetic vasomotor tone.²³ On the basis of the temporal changes in these hemodynamic parameters (Figure 2), experimental endotoxemia was divided into 3 phases.⁶ Phase I endotoxemia manifested an immediate and significant reduction in MSAP and the power density of vasomotor components of the SAP spectrum. Phase II exhibited a reversal of hypotension, along with a significant increase in sympathetic vasomotor outflow. Phase III was characterized by a significant secondary decrease in MSAP, HR, and the power density of the vasomotor components of SAP signals.

View larger version (27K): [in this window] [in a new window]   Figure 2. Temporal changes in MSAP, HR, and power density of vasomotor components of SAP spectrum in animals that received IV administration (at time 0; n=5 to 7 animals per group) of saline or LPS, given under NT or HS delivered at 8, 16, or 24 hours before treatment. Values are mean±SEM. *P<0.05 vs NT+saline group, #P<0.05 vs NT+LPS group in Scheffé multiple range analysis.

Twenty-four hours after HS, when upregulation of HSP70 expression in the ventrolateral medulla was optimal, the cardiovascular depression during phases II and III of endotoxemia was almost completely blunted. The same pretreatment, on the other hand, had no discernible effect on phase I endotoxemia. Interestingly, whereas bilateral microinjection of an antisense hsp70 oligonucleotide (50 pmol) into the RVLM significantly blunted the cardiovascular protection conferred by prior HS on phases II and III endotoxemia, treatment with sense or scrambled hsp70 oligonucleotide (50 pmol) was ineffective (Figure 3).

View larger version (24K): [in this window] [in a new window]   Figure 3. Temporal changes in MSAP, HR, and power density of vasomotor components of SAP spectrum in animals that received IV administration (at time 0; n=5 to 6 animals per group) of LPS, given under NT or 24 hours after HS, delivered alone or with additional bilateral microinjection of antisense (ASON) or sense (SON) hsp70 oligonucleotide (50 pmol) into RVLM. Note that results from animals that received microinjection of scrambled hsp70 oligonucleotide are not shown because they essentially duplicated those of SON-treated group. Values are mean±SEM. *P<0.05 vs NT+LPS group, #P<0.05 vs HS-24+LPS group in Scheffé multiple range analysis.

Hyperthermic HS Attenuated iNOS Upregulation in Ventrolateral Medulla During Experimental Endotoxemia
As exemplified by observations during phase III endotoxemia, the markedly upregulated iNOS mRNA (Figure 4A) or protein (Figure 4B) in the ventrolateral medull^5,6 was significantly attenuated 16 or 24 hours after HS, during which HSP70 expression was optimal. Comparable results were obtained during phase II endotoxemia (data not shown).

View larger version (33K): [in this window] [in a new window]   Figure 4. Representative gels for RT-PCR products of iNOS mRNA (inset A) or Western blot analysis of iNOS protein (inset B) and amount of iNOS mRNA or protein relative to GADPH (A) or ß-tubulin (B) detected from ventrolateral medulla 5 hours (phase III endotoxemia) after rats received IV saline or E. coli LPS (20 mg/kg) treatment, alone or with additional HS or NT, delivered 16, 24, or 48 hours before LPS treatment. Note that for clarity, only results on samples obtained 24 hours after NT were presented, although samples collected at all corresponding time intervals exhibited levels of iNOS mRNA or protein that were comparable to LPS-treated groups. Values are mean±SEM of quadruplicate analyses on samples pooled from 4 to 5 animals in each group. *P<0.05 vs sham-control group, #P<0.05 vs corresponding LPS group in Scheffé multiple range analysis.

B Decoy DNA Reversed Cardiovascular Depression or iNOS Upregulation in Ventrolateral Medulla During Experimental Endotoxemia
Microinjection of the double-stranded B decoy DNA (10 µg) bilaterally into the RVLM 24 hours before LPS treatment significantly reversed hypotension, bradycardia, and the decrease in power density of vasomotor components of the SAP spectrum (Figure 5A) or antagonized the upregulated iNOS mRNA (Figure 5B) or protein (Figure 5C) during phase II or III endotoxemia. Control microinjection of double-stranded DNA with a scrambled sequence (10 µg) was ineffective. The double-stranded B decoy DNA alone had no discernible effect on basal MSAP, HR, sympathetic vasomotor tone, or iNOS mRNA or protein expression.

View larger version (29K): [in this window] [in a new window]   Figure 5. Temporal changes in MSAP, HR, and power density of vasomotor components of SAP spectrum (A) or representative gels of iNOS mRNA (B) or protein (C) detected from ventrolateral medulla in animals that received IV administration (at time 0; n=5 to 6 animals per group) of saline or LPS (20 mg/kg), given alone or 24 hours after bilateral microinjection of B decoy or scrambled DNA (10 µg) into RVLM. Data shown in B and C are representative of 4 independent experiments. Values are mean±SEM *P<0.05 vs saline group, #P<0.05 vs LPS group in Scheffé multiple range analysis.

Activation of NF-B in Ventrolateral Medulla During Experimental Endotoxemia and Its Modulation by Hyperthermic HS
Electrophoresis mobility shift assay (Figure 6A, left panel) showed, contrary to saline, a significant increase in the association of NF-B with its consensus DNA oligonucleotide in nuclear extracts from the ventrolateral medulla 120 minutes after LPS treatment (phase II endotoxemia). Similar results were obtained during phase III endotoxemia (data not shown). Supershift experiments further revealed that the major NF-B family member activated during phase II or III endotoxemia was the p65/p50 heterodimer, because p65 or p50 but not c-Rel antiserum retarded the migration of proteins that interacted with the NF-B oligonucleotide (Figure 6A, right panel). Intriguingly, the increase in DNA binding activity of NF-B in the ventrolateral medulla was inhibited by prior HS (Figure 6A, left panel). This inhibition of an LPS-promoted increase in DNA binding activity of NF-B 24 hours after HS was reversed (Figure 6B) in animals that received treatment with antisense but not sense or scrambled hsp70 oligonucleotide (50 pmol).

View larger version (40K): [in this window] [in a new window]   Figure 6. Representative autoradiographs depicting NF-B DNA binding in nuclear extracts from ventrolateral medulla 120 minutes after animals received IV saline or LPS (20 mg/kg), alone or 16, 24, or 36 hours after HS (A, left panel), and in animals that received additional bilateral microinjection of artificial cerebrospinal fluid (aCSF) or antisense (ASON), sense (SON), or scrambled (SRON) hsp70 oligonucleotide (50 pmol) into RVLM 24 hours before LPS treatment (B). Also shown is NF-B DNA binding in nuclear extracts from ventrolateral medulla of LPS-treated animals preincubated in absence (untreated) or presence of antiserum against p50, p65, or c-Rel subunit of NF-B (A, right panel). Temporal changes in cytosolic and nuclear content of NF-B in ventrolateral medulla of animals after LPS treatment 24 hours after NT or HS are shown, respectively, in C and D. Values are mean±SEM of quadruplicate analyses on samples pooled from 4 to 5 animals in each group. *P<0.05 vs corresponding sham-control group in Scheffé multiple range analysis.

Nuclear Translocation of NF-B During Experimental Endotoxemia and Its Modulation by Hyperthermic HS
Immunoblot analysis revealed that expression of NF-B p65 protein in the cytosolic fraction of the ventrolateral medulla sample exhibited a significant decline 60 minutes after LPS administration, reaching 44±6% (n=5, P<0.05) of its initial value within 120 minutes (Figure 6C). Concomitantly, the progressively elevated NF-B p65 protein in the nuclear fraction reached 148±10% (n=6, P<0.05) of its initial value 120 minutes after induction of experimental endotoxemia. Both the decrease in cytosolic and increase in nuclear fraction of NF-B p65 expression detected 60 to 120 minutes after LPS treatment were attenuated 24 hours after HS treatment (Figure 6D).

IKK Activity or Degradation of IB in the Ventrolateral Medulla During Experimental Endotoxemia and Its Modulation by Hyperthermic HS
Figure 7A shows that LPS treatment led to an increase in IKK activity in the ventrolateral medulla during early phase II endotoxemia, as manifested by an increase in IB phosphorylation. HS, delivered to animals 24 hours before endotoxemia, discernibly depressed such an increase in IKK enzyme activity. There was nonetheless no discernible change in IKK protein expression under either NT or HS conditions. Western blot analysis also demonstrated that IB protein in the ventrolateral medulla was degraded by 36±5% (n=6, P<0.05) within 30 minutes, followed by a complete reexpression 120 minutes after LPS treatment (Figure 7B). This degradation of IB protein in the ventrolateral medulla was again significantly antagonized by prior HS.

View larger version (41K): [in this window] [in a new window]   Figure 7. Representative gels showing IKK activity, presented as level of phospho-I-B, in ventrolateral medulla of LPS-treated animals, 24 hours after NT or HS (A). Also shown are representative Western blots of I-B (inset) and amount of I-B protein relative to preinjection control detected from ventrolateral medulla of LPS-treated animals 24 hours after NT or HS (B). Values are mean±SEM of quadruplicate analyses on samples pooled from 4 to 5 animals in each group. *P<0.05 vs sham-control group in Scheffé multiple range analysis. C indicates sham control.

	Discussion

Top Abstract Introduction Methods Results Discussion References

 
Our laboratory reported previously that overproduction of NO after activation of iNOS⁵ and formation of peroxynitrite⁶ in the RVLM play a pivotal role in the reduction of sympathetic vasomotor outflow and arterial pressure during experimental endotoxemia. The present study further showed that HSP70 induced by hyperthermia HS conferred protection against this induced circulatory depression via an antagonism against iNOS mRNA and protein upregulation in the RVLM. We additionally demonstrated that this protective action of HSP70 may be related to suppression of NF-B activation through stabilization of IB via prevention of IKK activation and inhibition of IB degradation.

We demonstrated previously that HS-induced HSP70 confers cardiovascular protection against heatstroke by potentiating the baroreceptor reflex response²¹ via upregulation of glutamate receptors²² in the nucleus tractus solitarii, the terminal site of baroreceptor afferents in the caudal medulla.²⁵ The present study further demonstrated that HS also elicits cardiovascular protection against experimental endotoxemia by an upregulation of HSP70 in the RVLM, the final integration site for neural regulation of sympathetic vasomotor activity.⁵ Together, these observations suggest an important role for HSP70 in central autonomic control of circulation. It is intriguing that our results indicate that HS-induced HSP70 promoted cardiovascular protection by suppressing the surge in iNOS mRNA and protein expression during fatal endotoxemia. These results suggest that in addition to its actions as a molecular chaperone,²⁶ HSP70 may promote circulatory protection during endotoxemia via transcriptional regulation of iNOS gene induction.

The present results provide the first in vivo demonstration of the functional significance of NF-B activation in the manifestation of cardiovascular depression and its modulation by HS-induced HSP70 during endotoxemia. We found in the present study that both protein level and DNA binding activity of NF-B in the RVLM were significantly augmented during endotoxemia. That the major subunit activated in the RVLM revealed by supershift analysis was the p65/p50 heterodimer is consistent with reports^24,27 that p65/p50 complex is the predominant form of NF-B in the central nervous system. More importantly, the present results suggest that suppression of iNOS mRNA expression by HSP70 in the RVLM during endotoxemia may result from an attenuation of NF-B activation. We observed that prior HS treatment retarded nuclear translocation of the p65 subunit of NF-B and its DNA binding activity during endotoxemia in the RVLM. Observations with antisense hsp70 oligonucleotide treatment further established a causative relationship between HS-induced HSP70 expressed in the ventrolateral medulla and blockade of NF-B DNA binding activity. Functionally, blockade by the double-stranded B decoy DNA of NF-B binding to its cognate site, B element, in the RVLM also significantly attenuated the iNOS surge and cardiovascular depression during endotoxemia.

The present results revealed 2 possible mechanisms via which HS preconditioning may suppress NF-B activation during experimental endotoxemia. One mechanism involves inhibition of IKK activation, which was proposed^28,29 to underlie the inhibition by HS of NF-B activation in peripheral tissues during inflammatory responses. The present results also showed that hyperthermic HS significantly attenuated the increase in IKK activity at the ventrolateral medulla during endotoxemia that preceded nuclear translocation of NF-B. The second mechanism entails inhibition of cytoplasmic IB degradation.³⁰ HS preconditioning significantly blunted the transient reduction in IB level at the ventrolateral medulla during experimental endotoxemia. However, because HSP70 also enters the nucleus,³¹ the possibility of impeding NF-B nuclear translocation simply by competing for nuclear pore complexes during endotoxemia cannot be excluded.

HSP70 expression was detected in the ventrolateral medulla under basal conditions. We reasoned that this was not due to nonspecific stress to the animals, because HSP70 was not detected by the same antiserum in the nucleus tractus solitarii in sham-control or NT rats.²¹ We also noted that endogenously expressed HSP70 has been identified in normal rodent brain by Western blot³² or proteomic³³ analysis.

In conclusion, the present study revealed that upregulation of HSP70 by hyperthermic HS in the RVLM conferred protection against cardiovascular depression during experimental endotoxemia. We further demonstrated that this cardiovascular protective effect was exerted via blockade of NF-B activation, possibly through prevention of IKK activation and inhibition of IB degradation, leading to inhibition of iNOS upregulation in the ventrolateral medulla. We recognize that HSP70 is not the only HSP induced after thermal preconditioning or the only change to a live organism. As such, other HS-induced cellular mechanisms that may elicit cardiovascular protection during endotoxemia should not be overlooked.

	Acknowledgments

 
This study was supported by research grant VGHKS93–16 (Dr Julie Chan) from the Kaohsiung Veterans General Hospital and the Academic Excellence Program (89-B-FAO8-1-4) from the Ministry of Education (Dr Julie Chan and Dr Samuel Chan). Dr Samuel Chan is National Chair Professor of Neuroscience, appointed by the Ministry of Education, and Sun Yat-sen Research Chair Professor, appointed by National Sun Yat-sen University, Taiwan.

	Footnotes

 
The online-only Data Supplement, which contains additional information about Methods, is available with this article at http://www.circulationaha.org.

	References

Top Abstract Introduction Methods Results Discussion References

 
1. Parrillo JE. Pathogenetic mechanisms of septic shock. N Engl J Med. 1993; 328: 953–963.

2. Moncada S, Palmer RMJ, Higgs EA. Nitric oxide: physiology, pathophysiology and pharmacology. Pharmacol Rev. 1991; 43: 109–142.[Medline] [Order article via Infotrieve]

3. De Kimpe SJ, Kengatharan M, Thiemermann C, et al. The cell wall components peptidoglycan and lipoteichoic acid from Staphylococcus aureus act in synergy to cause shock and multiple organ failure. Proc Natl Acad Sci U S A. 1995; 92: 10359–10363.[Abstract/Free Full Text]

4. Chan SHH, Wang LL, Wang SH, et al. Differential cardiovascular responses to blockade of nNOS or iNOS in rostral ventrolateral medulla of the rat. Br J Pharmacol. 2001; 133: 606–614.[CrossRef][Medline] [Order article via Infotrieve]

5. Chan JYH, Wang SH, Chan SHH. Differential roles of iNOS and nNOS at rostral ventrolateral medulla during experimental endotoxemia in the rat. Shock. 2001; 15: 65–72.[Medline] [Order article via Infotrieve]

6. Chan SHH, Wang LL, Ou CC, et al. Contribution of peroxynitrite to fatal cardiovascular depression induced by overproduction of nitric oxide in rostral ventrolateral medulla of the rat. Neuropharmacology. 2002; 43: 889–898.[CrossRef][Medline] [Order article via Infotrieve]

7. Ross CA, Ruggiero DA, Park DH, et al. Tonic vasomotor control by the rostral ventrolateral medulla: effect of electrical or chemical stimulation of the area containing C1 adrenaline neurons on arterial pressure, heart rate, and plasma catecholamine and vasopressin. J Neurosci. 1984; 4: 474–494.[Abstract]

8. Xie QW, Kashiwabara Y, Nathan C. Role of transcription factor NF-B/Rel in induction of nitric oxide synthase. J Biol Chem. 1994; 269: 4705–4708.[Abstract/Free Full Text]

9. Baeuerle PA, Henkel T. Function and activation of NF-B in the immune system. Annu Rev Immunol. 1994; 12: 141–179.[Medline] [Order article via Infotrieve]

10. Maulik N, Goswami S, Galang N, et al. Differential regulation of Bcl-2, AP-1 and NF-B on cardiomyocyte apoptosis during myocardial ischemic stress adaptation. FEBS Lett. 1999; 443: 331–336.[CrossRef][Medline] [Order article via Infotrieve]

11. Armstead VE, Opentanova IL, Minchenko AG, et al. Tissue factor expression in vital organs during murine traumatic shock: role of transcription factors AP-1 and NF-B. Anesthesiology. 1999; 91: 1844–1852.[CrossRef][Medline] [Order article via Infotrieve]

12. Nathan C, Xie QW. Regulation of biosynthesis of nitric oxide. J Biol Chem. 1994; 269: 13725–13728.[Free Full Text]

13. Huang KC, Chen CW, Chen JC, et al. HMG-CoA reductase inhibitors inhibit inducible nitric oxide synthase gene expression in macrophages. J Biomed Sci. 2003; 10: 396–405.[Medline] [Order article via Infotrieve]

14. Pingle SC, Sanchez JF, Hallam DM, et al. Hypertonicity inhibits lipopolysaccharide-induced nitric oxide synthase expression in smooth muscle cells by inhibiting nuclear factor B. Mol Pharmacol. 2003; 63: 1238–1247.[Abstract/Free Full Text]

15. Feinstein DL, Galea E, Aquino DA, et al. Heat shock protein 70 suppresses astroglial-inducible nitric oxide synthase expression by decreasing NFB activation. J Biol Chem. 1996; 271: 17724–17732.[Abstract/Free Full Text]

16. Lindquist S, Craig EA. The heat-shock proteins. Annu Rev Genet. 1988; 22: 631–677.[CrossRef][Medline] [Order article via Infotrieve]

17. Hauser GJ, Dayao EK, Wasserloos K, et al. HSP induction inhibits iNOS mRNA expression and attenuates hypotension in endotoxin-challenged rats. Am J Physiol. 1996; 271: H2529–H2535.[Medline] [Order article via Infotrieve]

18. Lau SS, Griffin TM, Mestril R. Protection against endotoxemia by HSP70 in rodent cardiomyocytes. Am J Physiol. 2000; 278: H1439–H1445.

19. Wong HR, Ryan M, Wispe JR. The heat shock response inhibits inducible nitric oxide synthase gene expression by blocking I-B degradation and NF-B nuclear translocation. Biochem Biophys Res Commun. 1997; 231: 257–263.[CrossRef][Medline] [Order article via Infotrieve]

20. Meldrum KK, Burnett AL, Meng X, et al. Liposomal delivery of heat shock protein 72 into renal tubular cells blocks nuclear factor-B activation, tumor necrosis factor- production, and subsequent ischemia-induced apoptosis. Circ Res. 2003; 92: 293–299.[Abstract/Free Full Text]

21. Li PL, Chao YM, Chan SHH, et al. Potentiation of baroreceptor reflex response by heat shock protein 70 in nucleus tractus solitarii confers cardiovascular protection during heatstroke. Circulation. 2001; 103: 2114–2119.[Abstract/Free Full Text]

22. Chan SHH, Chang KF, Ou CC, et al. Up-regulation of glutamate receptors in nucleus tractus solitarii underlies potentiation of baroreceptor reflex by heat shock protein 70. Mol Pharmacol. 2002; 61: 1097–1104.[Abstract/Free Full Text]

23. Kuo TBJ, Yang CHH, Chan SHH. Selective activation of vasomotor component of SAP spectrum by nucleus reticularis ventrolateralis in rats. Am J Physiol. 1997; 272: H458–H492.

24. Yeh SH, Lin CH, Lee CF, et al. A requirement of nuclear factor-B activation in fear-potentiated startle. J Biol Chem. 2002; 277: 46720–46729.[Abstract/Free Full Text]

25. Ciriello J. Brainstem projections of aortic baroreceptor afferent fibers in the rat. Neurosci Lett. 1983; 36: 37–42.[CrossRef][Medline] [Order article via Infotrieve]

26. Hartl FU. Molecular chaperones in cellular protein folding. Nature. 1996; 381: 571–579.[CrossRef][Medline] [Order article via Infotrieve]

27. DeMeester SL, Buchman TG, Cobb FP. The heat shock paradox: NF-B determine cell fate? FASEB J. 2001; 15: 270–274.[Abstract/Free Full Text]

28. Kohn G, Wong HR, Bshesh K, et al. Heat shock inhibits TNF-induced ICAM-1 expression in human endothelial cells via I kappa kinase inhibition. Shock. 2002; 17: 91–97.[CrossRef][Medline] [Order article via Infotrieve]

29. Yoo CG, Lee S, Lee CT, et al. Anti-inflammatory effect of heat shock protein induction is related to stabilization of IB through preventing IB kinase activation in respiratory epithelial cells. J Immunol. 2000; 164: 5416–5423.[Abstract/Free Full Text]

30. Pritts TA, Wang Q, Sun X, et al. The stress response decreases NF-B activation in liver of endotoxemic mice. Shock. 2002; 18: 33–37.[CrossRef][Medline] [Order article via Infotrieve]

31. Dang CV, Lee WMF. Nuclear and nucleolar targeting sequence of c-erb-A, c-myb, N-myc, HSP70 and HIV tat proteins. J Biol Chem. 1989; 264: 18019–18023.[Abstract/Free Full Text]

32. Ren M, Leng Y, Jeong M, et al. Valproic acid reduces brain damage induced by transient focal cerebral ischemia in rats: potential roles of histone deacetylase inhibition and heat shock protein induction. J Neurochem. 2004; 89: 1358–1367.[CrossRef][Medline] [Order article via Infotrieve]

33. Dhodda VK, Sailor KA, Bowen KK, et al. Putative endogenous mediators of preconditioning-induced ischemic tolerance in rat brain by genomic and proteomics analysis. J Neurochem. 2004; 89: 73–89.[CrossRef][Medline] [Order article via Infotrieve]

This article has been cited by other articles:

				K. M. Stuhlmeier, J. Broll, and B. Iliev NF-KappaB Independent Activation of a Series of Proinflammatory Genes by Hydrogen Sulfide Experimental Biology and Medicine, November 1, 2009; 234(11): 1327 - 1338. [Abstract] [Full Text] [PDF]

				M. Choi, B. Salanova, S. Rolle, M. Wellner, W. Schneider, F. C. Luft, and R. Kettritz Short-Term Heat Exposure Inhibits Inflammation by Abrogating Recruitment of and Nuclear Factor-{kappa}B Activation in Neutrophils Exposed to Chemotactic Cytokines Am. J. Pathol., February 1, 2008; 172(2): 367 - 777. [Abstract] [Full Text] [PDF]

				K.-I. Tanaka, T. Namba, Y. Arai, M. Fujimoto, H. Adachi, G. Sobue, K. Takeuchi, A. Nakai, and T. Mizushima Genetic Evidence for a Protective Role for Heat Shock Factor 1 and Heat Shock Protein 70 against Colitis J. Biol. Chem., August 10, 2007; 282(32): 23240 - 23252. [Abstract] [Full Text] [PDF]

				J. Y. H. Chan, C. H. Y. Wu, C.-Y. Tsai, H.-L. Cheng, K.-Y. Dai, S. H. H. Chan, and A. Y. W. Chang Transcriptional up-regulation of nitric oxide synthase II by nuclear factor-{kappa}B at rostral ventrolateral medulla in a rat mevinphos intoxication model of brain stem death J. Physiol., June 15, 2007; 581(3): 1293 - 1307. [Abstract] [Full Text] [PDF]

				M. Sakamoto, T. Minamino, H. Toko, Y. Kayama, Y. Zou, M. Sano, E. Takaki, T. Aoyagi, K. Tojo, N. Tajima, et al. Upregulation of Heat Shock Transcription Factor 1 Plays a Critical Role in Adaptive Cardiac Hypertrophy Circ. Res., December 8, 2006; 99(12): 1411 - 1418. [Abstract] [Full Text] [PDF]

				A. Y. W. Chang, J. Y. H. Chan, J. L. J. Chou, F. C. H. Li, K.-Y. Dai, and S. H. H. Chan Heat shock protein 60 in rostral ventrolateral medulla reduces cardiovascular fatality during endotoxaemia in the rat J. Physiol., July 15, 2006; 574(2): 547 - 564. [Abstract] [Full Text] [PDF]

				J. Liu, K. W. L. Kam, G. H. Borchert, G. M. Kravtsov, H. J. Ballard, and T. M. Wong Further study on the role of HSP70 on Ca2+ homeostasis in rat ventricular myocytes subjected to simulated ischemia Am J Physiol Cell Physiol, February 1, 2006; 290(2): C583 - C591. [Abstract] [Full Text] [PDF]

				F. C. H. Li, J. Y. H. Chan, S. H. H. Chan, and A. Y. W. Chang In the Rostral Ventrolateral Medulla, the 70-kDa Heat Shock Protein (HSP70), but Not HSP90, Confers Neuroprotection against Fatal Endotoxemia via Augmentation of Nitric-Oxide Synthase I (NOS I)/Protein Kinase G Signaling Pathway and Inhibition of NOS II/Peroxynitrite Cascade Mol. Pharmacol., July 1, 2005; 68(1): 179 - 192. [Abstract] [Full Text] [PDF]

This Article Abstract Full Text (PDF) Data Supplement All Versions of this Article: 110/23/3560    most recent 01.CIR.0000143082.63063.33v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Chan, J. Y.H. Articles by Chan, S. H.H. Search for Related Content PubMed PubMed Citation Articles by Chan, J. Y.H. Articles by Chan, S. H.H. Pubmed/NCBI databases Compound via MeSH Substance via MeSH Related Collections Animal models of human disease Cell signalling/signal transduction