AJP - GI Watch the video to see how APS reaches out to developing nations.
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Am J Physiol Gastrointest Liver Physiol 291: G260-G266, 2006. First published March 30, 2006; doi:10.1152/ajpgi.00390.2005
0193-1857/06 $8.00
This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
291/2/G260    most recent
00390.2005v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Web of Science (11)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Szalay, L.
Right arrow Articles by Chaudry, I. H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Szalay, L.
Right arrow Articles by Chaudry, I. H.

INFLAMMATION/IMMUNITY/MEDIATORS

Androstenediol administration after trauma-hemorrhage attenuates inflammatory response, reduces organ damage, and improves survival following sepsis

László Szalay,* Tomoharu Shimizu,* Takao Suzuki, Ya-Ching Hsieh, Mashkoor A. Choudhry, Martin G. Schwacha, Kirby I. Bland, and Irshad H. Chaudry

Center for Surgical Research and Department of Surgery, University of Alabama at Birmingham, Alabama

Submitted 23 August 2005 ; accepted in final form 28 February 2006


   ABSTRACT
TOP
 ABSTRACT
MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 GRANTS
 REFERENCES
 
Although androstenediol (adiol or 5-androstene-3beta,17beta-diol), a metabolite of dehydroepiandrosterone (DHEA), has protective effects following trauma-hemorrhage (T-H), it remains unknown whether administration of adiol has any salutary effects on the inflammatory response and outcome following a combined insult of T-H and sepsis. Male rats underwent T-H shock [mean arterial pressure (MAP) 40 mmHg for 90 min] followed by resuscitation. Adiol (1 mg/kg body wt) or vehicle was administered at the end of resuscitation. Sepsis was induced by cecal ligation and puncture (CLP) at 20 h after T-H or sham operation. Five hours after CLP, plasma and tissue samples were analyzed for cytokines (IL-6 and IL-10), MPO, neutrophil chemotactic factor (CINC-3), and liver injury (alanine aminotransferase and lactate dehydrogenase). In another group of rats, the gangrenous cecum was removed at 10 h after CLP, the cavity was irrigated with warm saline and closed in layers, and mortality was recorded over 10 days. T-H followed by CLP produced a significant elevation in plasma IL-6 and IL-10 levels, enhanced neutrophil cell activation, and resulted in liver injury. Adiol administration prevented the increase in cytokine production, neutrophil cell activation, and attenuated liver injury. Moreover, rats subjected to the combined insult, receiving vehicle or adiol, had a 50% and 6% mortality, respectively. Since adiol administration suppresses proinflammatory cytokines, reduces liver damage, and decreases mortality after the combined insult of T-H and sepsis, this agent appears to be a novel adjunct to fluid resuscitation for decreasing T-H-induced septic complications and mortality.

liver enzymes; proinflammatory cytokines; neutrophils activation; myeloperoxidase

DESPITE NUMEROUS ADVANCES in intensive care medicine, sepsis and organ dysfunction remains the major cause of death in trauma patients as well as in patients following major surgery (6, 15, 37). A significant complicating factor following severe hemorrhage is the development of sepsis (5, 11, 23, 27). A growing body of data supports the notion that the prior hemorrhagic shock can significantly modulate the inflammatory response to a subsequent septic challenge. This is consistent with the "two-hit" model in which a major injury, such as hemorrhagic shock or trauma, acts as the "first hit" and sets up the host to manifest a suppressed or abnormal response to a secondary insult, such as the increased translocation of intestinal microbial or other such factors (8, 9, 13, 16).

The depressed organ perfusion and excessive production of proinflammatory mediators may play an important role in the development of multiple organ dysfunctions following hemorrhagic shock and sepsis (15, 21, 31). It has also been reported that systemic IL-6 levels increased after trauma-hemorrhage (T-H), and a sustained elevation in plasma IL-6 levels correlated with the evolving organ dysfunction (2, 9, 17, 38). Clinical data also confirmed that the increased IL-6 production is tied to a poor outcome (24).

Studies have demonstrated that the IL-6-mediated elevation in nitric oxide production is likely one of the detrimental effects induced by this cytokine. It has been reported that nitric oxide is a determinant of IL-6-mediated negative inotropic effects on the heart (10). Yu et al. (39) also demonstrated that IL-6 decreases cardiac contractility and enhances the synthesis of the inducible nitric oxide synthase (iNOS), resulting in an excess of nitric oxide production. In support of these findings, our studies have demonstrated that there was a close correlation between organ dysfunction and the elevation in plasma IL-6 levels (1–4, 13, 14, 17, 19, 38). Our recent study also indicated that the depressed cardiac function following T-H is associated with an increased in situ IL-6 production by the cardiomyocytes (38). The causal role of IL-6 in organ dysfunction has been further established by studies that showed that administration of anti-IL-6 antibodies improved organ function following T-H (34).

Androstenediol (adiol or 5-androstene-3beta,17beta-diol), a metabolite of dehydroepiandrosterone (DHEA), was shown to have greater protective capacity than DHEA on lethal bacterial infections and endotoxin shock (4). Additionally, it is reported that the conversion of DHEA leads to an increase in downstream effector hormones in macrophages, which may play an important role in local immunomodulation (25). Administration of adiol following T-H has also been shown to be effective in reducing plasma IL-6 production and improving cardiovascular and hepatic function (29, 30).

Although adiol has been reported to produce the above-mentioned salutary effects following T-H or endotoxin shock, it remains unknown whether its salutary effects are also evident in a two-hit model of T-H and sepsis. Therefore, the present study was undertaken to determine whether administration of adiol has any salutary effects on inflammatory mediators and on the survival of animals following T-H and sepsis. We hypothesized that adiol would have salutary effects on organ function and cytokine production even after a compound insult such as T-H and sepsis.


   MATERIALS AND METHODS
TOP
 ABSTRACT
 MATERIALS AND METHODS
RESULTS
 DISCUSSION
 GRANTS
 REFERENCES
 
Animals. Adult male (275–325 g) Sprague-Dawley rats (Charles River Laboratories, Wilmington, MA) were used in this study. All experiments were performed in adherence to the National Institutes of Health guidelines for the use of experimental animals and approved by the Institutional Animal Care and Use Committee of the University of Alabama at Birmingham.

Experimental groups. Following cannulation of the femoral arteries and a femoral vein, the animals were divided randomly into two groups, one group undergoing a T-H protocol and the second group in which only a sham operation was performed. These animals were then further divided into two subgroups as follows: at the end of resuscitation (or at the matching time point after sham operation), one subgroup was treated with androstenediol (adiol, 1 mg/kg body wt; Steraloids, Newport, RI), and the other group received the same volume of vehicle (vehicle, Intralipid, 1 ml/kg body wt; Sigma, St. Louis, MO). Twenty hours after the T-H or sham operation, the animals were anesthetized again, and cecal ligation and puncture was performed in both T-H and sham animals as described previously (5).

T-H procedure. A nonheparinized model of T-H, as previously described, was used in this study (17). Briefly, male Sprague-Dawley rats were fasted overnight before the experiment but allowed water ad libitum. Animals were anesthetized using isoflurane (Attane; Minrad, Bethlehem, PA) inhalation, followed by a 5-cm midline laparotomy to induce soft-tissue trauma. The abdomen was then closed in layers, and catheters were placed in both femoral arteries and the right femoral vein [polyethylene (PE)-50 tubing; Becton-Dickinson, Sparks, MD]. The animals were then restrained in a supine position, and the incision wounds were bathed with 1% lidocaine (Elkins-Sinn, Cherry Hill, NJ) throughout the surgical procedure to minimize postoperative pain. The rats were then allowed to awaken, following which they were bled rapidly to a mean arterial pressure (MAP) of 35–40 mmHg within 10 min. The time at which the animals could no longer maintain a MAP of 35–40 mmHg without infusing some fluid was defined as maximum bleed-out volume. The rats were maintained at that MAP until 40% of the shed blood volume was returned in the form of Ringer lactate. The animals were then resuscitated with four times the shed blood volume with Ringer lactate over 60 min. Adiol (1 mg/kg body wt iv) or vehicle was administered at the end of resuscitation. The catheters were then removed, the vessels were ligated, and the skin incisions were closed with sutures. Sham-operated animals underwent the same surgical procedure, which included the ligation of the femoral arteries and a femoral vein; however, neither hemorrhage nor resuscitation was performed. The animals were then returned to their cages and allowed food and water ad libitum. There was no mortality observed in this model of T-H shock.

Model of polymicrobial sepsis. At 20 h after the completion of fluid resuscitation or sham operation, the animals were again anesthetized with isoflurane, and polymicrobial sepsis was induced by cecal ligation and puncture (CLP) as described by Chaudry et al. (5). Briefly, following a 5-cm midline incision, the cecum was exposed and ligated just below the ileocecal valve. The cecum was then punctured twice with an 18-gauge needle and returned to the abdominal cavity. The abdominal cavity was then closed in layers, and the rats were resuscitated with saline solution subcutaneously (3 ml/100 g body wt). Animals were killed at 5 h following CLP to collect blood and tissue samples for further analysis. However, in the experiments in which survival was measured, the procedure of T-H and CLP was performed in a similar way, but 10 h after CLP, the gangrenous cecum was removed, the cavity was irrigated with warm saline, the abdominal incision was closed in layers, and mortality was recorded over a period of 10 days. The reason for this is to reduce the severity of the two-hit model; if we do not remove cecum, the model becomes very severe, and all animals will die within 24 h. No mortality is observed following T-H alone.

Plasma IL-6 and IL-10 levels. Plasma levels of IL-6 and IL-10 were determined using ELISA kits (R&D Systems, Minneapolis, MN) according to the manufacturer's instructions.

IL-6, IL-10, and iNOS mRNA expressions. Gene expressions of IL-6, IL-10, and iNOS were determined by quantitative real-time PCR as previously described (38). Total RNA was isolated from whole lung, heart, and liver samples using TRIzol reagent (Invitrogen, Carlsbad, CA) according to the manufacturer's protocol. The cDNA was generated from the total RNA samples by using TaqMan reverse transcription reagents (Applied Biosystems, Foster City, CA). Each real-time PCR reaction was performed in a 10-µl reaction mixture containing 20 ng of cDNA, 2x PCR master mix (Applied Biosystems), and each probe and primer set. TaqMan gene expression assays (Applied Biosystems) for IL-6, IL-10, and iNOS, were purchased as probe and primer sets. The reaction mixture was denatured for one cycle of 2 min at 50°C and 10 min at 95°C and incubated for 40 cycles (denaturing for 15 sec at 95°C and annealing and extending for 1 min at 60°C) using the ABI Prism 7900HT (Applied Biosystems). All samples were tested in triplicate, and average values were used for quantification. rRNA (18S) was used as an endogenous control. The comparative threshold cycle (CT) method was used for quantification of gene expression. Analysis was performed using SDS v2.2 software (Applied Biosystems) according to the manufacturer's instruction. The normalized data was used and expressed as percent of the CLP + vehicle group, which was considered as 1.0.

Myeloperoxidase activity. MPO activity, an indicator of neutrophil cell accumulation, was determined from lung and heart samples collected at 5 h after CLP. Each sample was homogenized in a 10x volume of solution containing 0.5% (wt/vol) hexadecyltrimethyl-ammonium bromide dissolved in 10 mM potassium phosphate buffer (pH 7) and centrifuged for 30 min at 20,000 g at 4°C. An aliquot of the supernatant was then allowed to react with a solution of tetramethylbenzidine (1.6 mM) and hydrogen peroxide (0.1 mM). The rate of change in absorbance was measured spectrophotometrically at 650 nm, and MPO activity was defined as units per gram of tissue protein. Protein concentration was determined using a Bio-Rad assay kit (Bio-Rad, Hercules, CA).

Cytokine-induced neutrophil chemoatractant-3 levels. Tissue levels of cytokine-induced neutrophil chemoatractant-3 (CINC-3) were determined using an ELISA kit (R&D Systems) according to the manufacturer's instructions. Tissue concentration of CINC-3 was defined as nanograms of CINC-3 per gram of tissue protein.

Alanine aminotransferase and lactate dehydrogenase assays. Plasma samples were examined for alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) levels. Measurements were performed spectrophotometrically by using diagnostic kits from Sigma according to the manufacturer's descriptions.

Statistical analysis. Values are means ± SE of 5–6 animals/group. For survival studies, 16 animals were used for each group. Statistical differences between groups were calculated either by one-way analysis of variance (ANOVA), followed by a Fisher LSD post hoc test, or by a Fisher exact test. Differences were considered significant if P < 0.05.


   RESULTS
TOP
 ABSTRACT
 MATERIALS AND METHODS
 RESULTS
DISCUSSION
 GRANTS
 REFERENCES
 
Cytokine production and iNOS expression. Five hours after CLP, animals of the T-H + vehicle + CLP group had significantly higher plasma IL-6 and IL-10 levels compared with CLP + vehicle (Figs. 1A and 2A). In addition, the IL-6 and IL-10 mRNA expressions were also elevated following a combined insult of T-H and CLP in the liver, heart, and lung in the vehicle group (Figs. 1, B–D, and 2, B–D). The adiol-treated T-H group subjected to CLP showed attenuated plasma IL-6 and IL-10 levels similar to shams. In addition, mRNA transcripts of the cytokines in this group also remained at a significantly lower level in the heart and the lung. In the liver, the IL-6 transcription was also reduced significantly, but the reduction in the IL-10 mRNA content did not reach statistical difference.


Figure 1
View larger version (17K):
[in this window]
[in a new window]
  		Fig. 1. Effects of androstenediol (adiol) administration on IL-6 production at 5 h after cecal ligation and puncture (CLP) or trauma-hemorrhage (T-H) + CLP. A: plasma levels of IL-6. B–D: IL-6 mRNA expressions in the liver (B), heart (C), and lung (D). The mRNA level of each CLP + vehicle (Veh) group is considered 1.0; mRNA values in the other three groups are expressed relative to the corresponding CLP + Veh; n = 6 animals/group. *P < 0.05 vs. corresponding CLP alone group. #P < 0.05 vs. corresponding Veh-treated group. RQ, respiratory quotient.

 

Figure 2
View larger version (19K):
[in this window]
[in a new window]
  		Fig. 2. Effects of adiol administration on IL-10 production at 5 h after CLP or T-H + CLP. A: plasma levels of IL-10. B–D: IL-10 mRNA expressions in the liver (B), heart (C), and lung (D). The mRNA level of each CLP + Veh group is considered 1.0; mRNA values in the other three groups are expressed relative to the corresponding CLP + Veh; n = 6 animals/group. *P < 0.05 vs. corresponding CLP alone group. #P < 0.05 vs. corresponding Veh-treated group.

 
In the group of animals not undergoing T-H prior to CLP, adiol did not induce significant alterations in plasma IL-6 or IL-10 levels. In the lung, IL-6 mRNA expression was significantly reduced in the adiol-treated T-H group subjected to sepsis compared with CLP group receiving vehicle. The expression of IL-6 and IL-10 mRNA in the liver, heart, and lung were not affected by adiol treatment.

Similar to the cytokine release, iNOS expression in liver, heart, and lung was also significantly upregulated following T-H and subsequent CLP. Adiol administration following T-H significantly prevented the increase of iNOS mRNA expression in the liver, heart, and lung (Fig. 3, A–C).


Figure 3
View larger version (11K):
[in this window]
[in a new window]
  		Fig. 3. Effects of adiol administration on iNOS mRNA expressions in the liver (A), heart (B), and lung (C) at 5 h after CLP. The mRNA level of the CLP + Veh group is considered 1.0; other mRNA values are expressed relative to this group; n = 6 animals/group. *P < 0.05 vs. corresponding CLP alone group. #P < 0.05 vs. corresponding Veh-treated group.

 
Neutrophil accumulation in the lung and the heart. MPO activity, an indicator of neutrophil accumulation, was determined in the lung and heart samples. Five hours after CLP, animals in the T-H group receiving vehicle following T-H had significantly higher heart and lung MPO levels compared with CLP animals receiving vehicle and not undergoing prior T-H (Fig. 4, A and B). In addition, adiol treatment significantly reduced MPO levels in both lung and heart following a combined insult of T-H and CLP. Adiol did not influence MPO activity significantly in the CLP group not subjected to prior T-H.


Figure 4
View larger version (16K):
[in this window]
[in a new window]
  		Fig. 4. Effects of adiol treatment on neutrophil accumulation in the heart (A) and lung (B). Tissue myeloperoxidase activities were measured at 5 h after CLP; n = 6 animals/group. *P < 0.05 vs. corresponding CLP alone group. #P < 0.05 vs. corresponding Veh-treated group.

 
Neutrophil CINC-3 in the liver, lung, and heart. There was a significant increase in CINC-3 levels in the liver and the heart following T-H and CLP (Fig. 5, A and B). Similarly, lung CINC-3 levels also displayed a trend for enhanced production, but the increase in lung CINC-3 levels following a combined insult of T-H and CLP did not reach statistical significance (Fig. 5C). Treatment of rats with adiol prevented the increase in CINC-3.


Figure 5
View larger version (13K):
[in this window]
[in a new window]
  		Fig. 5. Effect of adiol treatment on tissue cytokine-induced neutrophil chemoattractant (CINC-3) levels in the liver (A), heart (B), and lung (C) at 5 h after CLP; n = 5–6 animals/group. *P < 0.05 vs. corresponding CLP alone group.

 
Liver injury. Following the combined insult of T-H and CLP, plasma LDH (Fig. 6A) and ALT (Fig. 6B) levels were significantly higher than in the groups receiving CLP alone. Plasma LDH was reduced significantly in the adiol-treated T-H group subjected to CLP; however, it remained at a higher level than the levels in the adiol + CLP group. A small but insignificant reduction in ALT level was also observed in the former group. Furthermore, adiol did not influence plasma LDH and ALT levels in animals receiving CLP alone in the absence of T-H.


Figure 6
View larger version (11K):
[in this window]
[in a new window]
  		Fig. 6. Effects of adiol treatment on plasma lactate dehydrogenase (LDH) (A) and alanine aminotransferase (ALT) (B) levels following CLP or T-H + CLP. Plasma samples were collected 5 h after CLP; n = 6 animals/group. *P < 0.05 vs. corresponding CLP alone group. #P < 0.05 vs. corresponding Veh-treated group.

 
Survival rate. CLP alone resulted in no mortality within the 10-day survival observation period (Fig. 7). The animals in the T-H group subjected to CLP had a mortality rate of 50% within 3 days after CLP. Adiol treatment, however, improved the survival rate in the combined model of T-H + CLP, and the survival rate was similar to that observed in the CLP group that did not undergo prior T-H.


Figure 7
View larger version (7K):
[in this window]
[in a new window]
  		Fig. 7. Effect of adiol treatment on survival. Rats were monitored for a period of 10 days following the CLP and excision, and survival in each group was assessed at 24 h intervals; n = 16 animals/group. #P < 0.05 vs. T-H + adiol + CLP.

 

   DISCUSSION
TOP
 ABSTRACT
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
GRANTS
 REFERENCES
 
The present results indicate that adiol administration reduces cardiac IL-6 and iNOS gene expressions following the combined insult of T-H and sepsis. Since IL-6 plays a major role in the development of multiple organ failure following hemorrhage, it is possible that adiol treatment would improve the survival of animals subjected to T-H and sepsis, via reducing IL-6. The present results also indicate that the higher IL-6 levels were associated with higher mortality and that lower levels of IL-6 with adiol administration were associated with decreased mortality. Previous studies from our laboratory have shown that Kupffer cells are the major source of circulating IL-6 following T-H as well as sepsis (3, 22). In accordance with these data, Kupffer cells isolated from posthemorrhaged or postseptic animals also showed a markedly higher IL-6 production in vitro (16). Our preliminary study indicates that the Kupffer cells isolated from T-H rats subjected to subsequent sepsis also showed an increased IL-6 production, which was decreased to sham levels by administration of adiol following T-H (unpublished observations). Our recent study has shown that following T-H, IL-6 production by cardiomyocytes locally can also significantly produce cardiac depression under those conditions (38). The present study shows that IL-6 mRNA levels in the heart and plasma IL-6 levels also increased significantly in the group subjected to T-H and sepsis. Studies have also shown that high IL-6 levels decreased cardiac contractility, and this was associated with an enhanced synthesis of iNOS and increased NO production by the myocytes (39). In this regard, the significance of iNOS induction and the deleterious effects of excessive NO on contractile function has been well-demonstrated (28, 33). In addition, the contribution of iNOS in the induction of organ injury following hemorrhage or septic insult has also been suggested (5, 7, 18, 31, 32). Our recent findings revealed that adiol administration following T-H decreased plasma IL-6 levels and improved cardiac function (29). However, that study did not examine the effects of adiol following a combined insult of T-H and induction of sepsis, which was examined in this study. Previous studies from our laboratory have also shown a correlation between the adiol-mediated attenuation of iNOS expression and reduction in hepatic injury (30). Although the significance of iNOS induction still remains unclear, it is likely that the observed downregulation of iNOS gene expression in the liver, heart, and lung in this study contributes to the adiol-mediated salutary effects on organ functions following T-H and the induction of subsequent sepsis.

The results indicate that MPO activity in the heart and lung increased significantly in the combined model of T-H and sepsis, indicating neutrophil activation under those conditions. In contrast, the MPO activity was unaffected in the heart as well as in the lung at 5 h after the onset of sepsis. However, since we did not use normal rats for comparison, it could be argued that the MPO was indeed increased with sepsis alone. Although this may be so, the fact remains that adiol had no effect on heart or lung MPO in the model of sepsis alone, thereby suggesting that 5 h may be too early for sepsis in itself to produce neutrophil activation. Further support for the notion that 5 h after the onset of sepsis without prior T-H did not produce any significant adverse effects comes from the finding that liver LDH and ALT levels were also not increased at that interval without or with adiol administration. These results are in accordance with our previous results that showed that the liver enzymes were not increased at 5 h after CLP (35). Furthermore, CINC-3 was not affected significantly at 5 h after CLP, and although CINC-3 levels were detected in the liver and lung, the levels were not affected by adiol, again suggesting that these are normal levels of CINC-3 in the liver and lung. Altogether, the results as summarized in this manuscript clearly suggest that adiol prevents both the inflammatory response and organ dysfunction following a combined insult of T-H and CLP.

A number of studies have also shown that IL-10 is capable of inhibiting the production of macrophage-derived proinflammatory cytokines. Since ablation of Kupffer cells by gadolinium chloride also decreased circulating IL-10 levels following T-H, it indicates that Kupffer cells also represent the major cellular source of elevated circulating IL-10 levels under those conditions (26). In view of these results, it can be suggested that adiol-induced salutary effects are mediated, at least in part, by a decreased cytokine production by Kupffer cells. Although the precise mechanism responsible for the salutary effects of adiol on cytokine production remains unknown, it is possible that adiol mediates its salutary effects via the estrogen receptor(s). In this regard, studies have indicated that estrogen-like activity of adiol is observed at physiological concentration in breast cancer cells (20). Furthermore, adiol causes an increase in estrogen receptor-dependent beta-galactosidase activity in yeast (20). Previous studies from our laboratory have shown that proestrus female mice, which have high circulating levels of estrogen and progesterone, have a significantly lower mortality following T-H and the induction of subsequent sepsis (9) or sepsis alone (40) than male counterparts. Moreover, the salutary effects of DHEA (a parent compound of adiol) on organ functions are abolished if the estrogen receptor antagonist ICI 182,780 is administered with DHEA (12). This suggests that in addition to adiol, DHEA also mediates its effects via the estrogen receptor(s) (12).

Another potential mechanism of the salutary effect of adiol could be the activation of peroxisomes. Waxman (36) has demonstrated that adiol can activate peroxisome proliferator gamma (PPAR-{gamma}). Activated PPAR-{gamma} could reduce organ injury in T-H shock and systemic inflammation in polymicrobial sepsis (1, 41). Our recent study indicated that adiol treatment following T-H activated PPAR-{gamma}, and treatment with PPAR-{gamma} antagonist prevented the salutary effects of adiol under those conditions (unpublished observations). Thus it is likely that adiol produces its salutary effects via PPAR-{gamma}; however, more studies are needed to delineate the precise mechanism of the salutary effects of this compound following T-H and/or sepsis.

Previous studies have shown that CLP alone induces an inflammatory response at a time point later than 5 h, causes organ dysfunction, and potentiates the inflammatory response following T-H (9, 13, 16, 35). However, CLP-induced inflammatory response in the absence of T-H was not observed at an early time point (i.e., 5 h after CLP). The primary reason to perform CLP in this study was not to determine the effect of CLP on the inflammatory response or organ function, rather its potential effect on the post-T-H inflammatory response and organ function. Since at 5 h there was no effect of CLP on inflammatory response, we selected this time point to be optimum to differentiate between the responses induced by CLP and T-H alone as well as the responses induced in the event that the two, CLP and T-H, are combined. Thus measurement at 5 h after CLP has allowed us to determine the effect of CLP on the T-H-induced inflammatory response and organ function and whether those potential effects are prevented in adiol-treated animals.

Our studies suggest that T-H followed by CLP produced a significant elevation in plasma IL-6 and IL-10 levels, enhanced neutrophil infiltration in lung and heart, and caused liver injury. These alterations in inflammatory mediators accompanied with an increase in mortality following T-H and sepsis. Adiol administration suppresses proinflammatory cytokines and decreases mortality rates following T-H and sepsis. The improved survival from sepsis by adiol treatment following T-H appears to be due to decreased organ damage and improved organ functions. We found that these improvements are accompanied with a reduction in IL-6 and/or iNOS production. Collectively, these findings suggest a relationship between increased inflammatory mediators and mortality following T-H and sepsis. Nevertheless, more studies are required to delineate the mechanism by which the decrease in inflammatory response results in an increase in antimicrobial defense. Furthermore, whether adiol modulation of global inflammatory response or any particular chemokine or cytokine is responsible for the salutary effects remains to be established. In summary, although the precise mechanism of the salutary effects of adiol requires further investigations, our studies suggest that adjuvant use of adiol following T-H appears to be a novel approach for reducing the mortality rates following T-H and the occurrence of subsequent sepsis.


   GRANTS
TOP
 ABSTRACT
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 GRANTS
REFERENCES
 
This study was supported by National Institute of General Medical Sciences Grant R37-GM-39519.


   FOOTNOTES
 

Address for reprint requests and other correspondence: I. H. Chaudry, Center for Surgical Research, Univ. of Alabama at Birmingham, 1670 Univ. Boulevard, Volker Hall, Rm. G094, Birmingham, AL 35294-0019 (e-mail: Irshad.Chaudry{at}ccc.uab.edu)

The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

* L. Szalay and T. Shimizu contributed equally to this work. Back


   REFERENCES
TOP
 ABSTRACT
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 GRANTS
 REFERENCES
 

  1. Abdelrahman M, Collin M, and Thiemermann C. The peroxisome proliferator-activated receptor-{gamma} ligand 15-deoxy{delta}12,14 prostaglandin J2 reduces the organ injury in hemorrhagic shock. Shock 22: 555–561, 2004.[CrossRef][Web of Science][Medline]
  2. Angele MK, Schwacha MG, Ayala A, and Chaudry IH. Effect of gender and sex hormones on immune responses following shock. Shock 14: 81–90, 2000.[Web of Science][Medline]
  3. Ayala A, O'Neill PJ, Uebele SA, Herdon CD, and Chaudry IH. Mechanism of splenic immunosuppression during sepsis: key role of Kupffer cell mediators. J Trauma 42: 882–888, 1997.[Web of Science][Medline]
  4. Ben-Nathan D, Padgett DA, and Loria RM. Androstenediol and dehydroepiandrosterone protect mice against lethal bacterial infections and lipopolysaccharide toxicity. J Med Microbiol 48: 425–431, 1999.[Abstract/Free Full Text]
  5. Chaudry IH, Wichterman KA, and Baue AE. Effect of sepsis on tissue adenine nucleotide levels. Surgery 85: 205–211, 1979.[Web of Science][Medline]
  6. Cobb JP, Brownstein BH, Watson MA, Shannon WD, Laramie JM, Qiu Y, Stormo GD, Morrissey JJ, Buchman TG, Karl IE, and Hotchkiss RS. Injury in the era of genomics. Shock 15: 165–170, 2001.[Web of Science][Medline]
  7. Collins JL, Vodovotz Y, Hierholzer C, Villavicencio RT, Liu S, Alber S, Gallo D, Stolz DB, Watkins SC, Godfrey A, Gooding W, Kelly E, Peitzman AB, and Billiar TR. Characterization of the expression of inducible nitric oxide synthase in rat and human liver during hemorrhagic shock. Shock 19: 117–122, 2003.[CrossRef][Web of Science][Medline]
  8. Deitch EA. Animal models of sepsis and shock: a review and lessons learned. Shock 9: 1–11, 1998.[Web of Science][Medline]
  9. Diodato MD, Knoferl MW, Schwacha MG, Bland KI, and Chaudry IH. Gender differences in the inflammatory response and survival following haemorrhage and subsequent sepsis. Cytokine 14: 162–169, 2001.[CrossRef][Web of Science][Medline]
  10. Finkel MS, Oddis CV, Jacob TD, Watkins SC, Hattler BG, and Simmons RL. Negative inotropic effects of cytokines on the heart mediated by nitric oxide. Science 257: 387–389, 1992.[Abstract/Free Full Text]
  11. Jacob S, Weizel H, Gordon E, Korman H, Schweinburg F, Frank H, and Fine J. Bacterial action in development of irreversibility to transfusion in hemorrhagic shock in the dog. Am J Physiol 179: 523–531, 1954.[Free Full Text]
  12. Jarrar D, Wang P, Cioffi WG, Bland KI, and Chaudry IH. Mechanisms of the salutary effects of dehydroepiandrosterone after trauma-hemorrhage: direct or indirect effects on cardiac and hepatocellular functions? Arch Surg 135: 416–422, 2000.[Abstract/Free Full Text]
  13. Jarrar D, Wang P, Song GY, Cioffi WG, Bland KI, and Chaudry IH. Inhibition of tyrosine kinase signaling after trauma-hemorrhage: a novel approach for improving organ function and decreasing susceptibility to subsequent sepsis. Ann Surg 231: 399–407, 2000.[CrossRef][Web of Science][Medline]
  14. Jarrar D, Wang P, Song GY, Knoferl MW, Cioffi WG, Bland KI, and Chaudry IH. Metoclopramide: a novel adjunct for improving cardiac and hepatocellular functions after trauma-hemorrhage. Am J Physiol Endocrinol Metab 278: E90–E95, 2000.[Abstract/Free Full Text]
  15. Kher A, Wang M, Tsai BM, Pitcher JM, Greenbaum ES, Nagy RD, Patel KM, Wairiuko GM, Markel TA, and Meldrum DR. Sex differences in the myocardial inflammatory response to acute injury. Shock 23: 1–10, 2005.[Web of Science][Medline]
  16. Knoferl MW, Diodato MD, Schwacha MG, Cioffi WG, Bland KI, and Chaudry IH. Cyclooxygenase-2-mediated regulation of Kupffer cell interleukin-6 production following trauma-hemorrhage and subsequent sepsis. Shock 16: 479–483, 2001.[Web of Science][Medline]
  17. Kuebler JF, Jarrar D, Bland KI, Rue L III, Wang P, and Chaudry IH. Progesterone administration after trauma and hemorrhagic shock improves cardiovascular responses. Crit Care Med 31: 1786–1793, 2003.[CrossRef][Web of Science][Medline]
  18. Menezes JM, Hierholzer C, Watkins SC, Billiar TR, Peitzman AB, and Harbrecht BG. The modulation of hepatic injury and heat shock expression by inhibition of inducible nitric oxide synthase after hemorrhagic shock. Shock 17: 13–18, 2002.[CrossRef][Web of Science][Medline]
  19. Mizushima Y, Wang P, Jarrar D, Cioffi WG, Bland KI, and Chaudry IH. Estradiol administration after trauma-hemorrhage improves cardiovascular and hepatocellular functions in male animals. Ann Surg 232: 673–679, 2000.[CrossRef][Web of Science][Medline]
  20. Nephew KP, Sheeler CQ, Dudley MD, Gordon S, Nayfield SG, and Khan SA. Studies of dehydroepiandrosterone (DHEA) with the human estrogen receptor in yeast. Mol Cell Endocrinol 143: 133–142, 1998.[CrossRef][Web of Science][Medline]
  21. Oberholzer A, Souza SM, Tschoeke SK, Oberholzer C, Abouhamze A, Pribble JP, and Moldawer LL. Plasma cytokine measurements augment prognostic scores as indicators of outcome in patients with severe sepsis. Shock 23: 488–493, 2005.[CrossRef][Web of Science][Medline]
  22. O'Neill PJ, Ayala A, Wang P, Ba ZF, Morrison MH, Schultze AE, Reich SS, and Chaudry IH. Role of Kupffer cells in interleukin-6 release following trauma-hemorrhage and resuscitation. Shock 1: 43–47, 1994.[Web of Science][Medline]
  23. Ravin HA, Rowley D, Jenkins C, and Fine J. On the absorption of bacterial endotoxin from the gastro-intestinal tract of the normal and shocked animal. J Exp Med 112: 783–792, 1960.[Abstract]
  24. Roumen RM, Hendriks T, van der Ven-Jongekrijg J, Nieuwenhuijzen GA, Sauerwein RW, van der Meer JW, and Goris RJ. Cytokine patterns in patients after major vascular surgery, hemorrhagic shock, and severe blunt trauma Relation with subsequent adult respiratory distress syndrome and multiple organ failure. Ann Surg 218: 769–776, 1993.[Web of Science][Medline]
  25. Schmidt M, Kreutz M, Loffler G, Scholmerich J, and Straub RH. Conversion of dehydroepiandrosterone to downstream steroid hormones in macrophages. J Endocrinol 164: 161–169, 2000.[Abstract]
  26. Schneider CP, Schwacha MG, and Chaudry IH. The role of interleukin-10 in the regulation of the systemic inflammatory response following trauma-hemorrhage. Biochim Biophys Acta 1689: 22–32, 2004.[Medline]
  27. Schweinburg FB, Seligman AM, and Fine J. Transmural migration of intestinal bacteria; a study based on the use of radioactive Escherichia coli. N Engl J Med 242: 747–751, 1950.[Web of Science][Medline]
  28. Shah AM, Prendergast BD, Grocott-Mason R, Lewis MJ, and Paulus WJ. The influence of endothelium-derived nitric oxide on myocardial contractile function. Int J Cardiol 50: 225–231, 1995.[CrossRef][Web of Science][Medline]
  29. Shimizu T, Choudhry MA, Szalay L, Rue LW 3rd, Bland KI, and Chaudry IH. Salutary effects of androstenediol on cardiac function and splanchnic perfusion after trauma-hemorrhage. Am J Physiol Regul Integr Comp Physiol 287: R386–R390, 2004.[Abstract/Free Full Text]
  30. Shimizu T, Szalay L, Choudhry MA, Schwacha MG, Rue LW, 3rd Bland KI, and Chaudry IH. Mechanism of salutary effects of androstenediol on hepatic function after trauma-hemorrhage: role of endothelial and inducible nitric oxide synthase. Am J Physiol Gastrointest Liver Physiol 288: G244–G250, 2005.[Abstract/Free Full Text]
  31. Szabo C and Billiar TR. Novel roles of nitric oxide in hemorrhagic shock. Shock 12: 1–9, 1999.[Medline]
  32. Szabo C and Thiemermann C. Invited opinion: role of nitric oxide in hemorrhagic, traumatic, and anaphylactic shock and thermal injury. Shock 2: 145–155, 1994.[Web of Science][Medline]
  33. Tatsumi T, Matoba S, Kawahara A, Keira N, Shiraishi J, Akashi K, Kobara M, Tanaka T, Katamura M, Nakagawa C, Ohta B, Shirayama T, Takeda K, Asayama J, Fliss H, and Nakagawa M. Cytokine-induced nitric oxide production inhibits mitochondrial energy production and impairs contractile function in rat cardiac myocytes. J Am Coll Cardiol 35: 1338–1346, 2000.[Abstract/Free Full Text]
  34. Toth B, Yokoyama Y, Schwacha MG, George RL, Rue LW, 3rd Bland KI, and Chaudry IH. Insights into the role of interleukin-6 in the induction of hepatic injury after trauma-hemorrhagic shock. J Appl Physiol 97: 2184–2189, 2004.[Abstract/Free Full Text]
  35. Wang P, Ba ZF, and Chaudry IH. Hepatic extraction of indocyanine green is depressed early in sepsis despite increased hepatic blood flow and cardiac output. Arch Surg 126: 219–224, 1991.[Abstract/Free Full Text]
  36. Waxman DJ. Role of metabolism in the activation of dehydroepiandrosterone as a peroxisome proliferator. J Endocrinol 150, Suppl: S129–S147, 1996.[CrossRef][Web of Science][Medline]
  37. Wu LL, Tang C, and Liu MS. Altered phosphorylation and calcium sensitivity of cardiac myofibrillar proteins during sepsis. Am J Physiol Regul Integr Comp Physiol 281: R408–R416, 2001.[Abstract/Free Full Text]
  38. Yang S, Zheng R, Hu S, Ma Y, Choudhry MA, Messina JL, Rue LW III, Bland KI, and Chaudry IH. Mechanism of cardiac depression after trauma-hemorrhage: increased cardiomyocyte IL-6 and effect of sex steroids on IL-6 regulation and cardiac function. Am J Physiol Heart Circ Physiol 287: H2183–H2191, 2004.[Abstract/Free Full Text]
  39. Yu X, Kennedy RH, and Liu SJ. JAK2/STAT3, not ERK1/2, mediates interleukin-6-induced activation of inducible nitric-oxide synthase and decrease in contractility of adult ventricular myocytes. J Biol Chem 278: 16304–16309, 2003.[Abstract/Free Full Text]
  40. Zellweger R, Wichmann MW, Ayala A, Stein S, DeMaso CM, and Chaudry IH. Females in proestrus state maintain splenic immune functions and tolerate sepsis better than males. Crit Care Med 25: 106–110, 1997.[CrossRef][Web of Science][Medline]
  41. Zingarelli B and Cook JA. Peroxisome proliferator-activated receptor-{gamma} is a new therapeutic target in sepsis and inflammation. Shock 23: 393–399, 2005.[CrossRef][Web of Science][Medline]



This article has been cited by other articles:


Home page
 Am. J. Physiol. Heart Circ. Physiol.Home page
T. A. Markel, Y. Wang, J. L. Herrmann, P. R. Crisostomo, M. Wang, N. M. Novotny, C. M. Herring, J. Tan, T. Lahm, and D. R. Meldrum
VEGF is critical for stem cell-mediated cardioprotection and a crucial paracrine factor for defining the age threshold in adult and neonatal stem cell function
Am J Physiol Heart Circ Physiol, December 1, 2008; 295(6): H2308 - H2314.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Regul. Integr. Comp. Physiol.Home page
T. A. Markel, P. R. Crisostomo, M. Wang, Y. Wang, T. Lahm, N. M. Novotny, J. Tan, and D. R. Meldrum
TNFR1 signaling resistance associated with female stem cell cytokine production is independent of TNFR2-mediated pathways
Am J Physiol Regulatory Integrative Comp Physiol, October 1, 2008; 295(4): R1124 - R1130.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Regul. Integr. Comp. Physiol.Home page
T. A. Markel, M. Wang, P. R. Crisostomo, M. C. Manukyan, J. A. Poynter, and D. R. Meldrum
Neonatal stem cells exhibit specific characteristics in function, proliferation, and cellular signaling that distinguish them from their adult counterparts
Am J Physiol Regulatory Integrative Comp Physiol, May 1, 2008; 294(5): R1491 - R1497.
[Abstract] [Full Text] [PDF]

Home page
 J. Leukoc. Biol.Home page
J. L. Sperry and J. P. Minei
Gender dimorphism following injury: making the connection from bench to bedside
J. Leukoc. Biol., March 1, 2008; 83(3): 499 - 506.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Gastrointest. Liver Physiol.Home page
T. A. Markel, P. R. Crisostomo, M. Wang, C. M. Herring, and D. R. Meldrum
Activation of individual tumor necrosis factor receptors differentially affects stem cell growth factor and cytokine production
Am J Physiol Gastrointest Liver Physiol, October 1, 2007; 293(4): G657 - G662.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Pharmacol.Home page
M. Xiao, C. E. Inal, V. I. Parekh, C.-M. Chang, and M. H. Whitnall
5-Androstenediol Promotes Survival of {gamma}-Irradiated Human Hematopoietic Progenitors through Induction of Nuclear Factor-{kappa}B Activation and Granulocyte Colony-Stimulating Factor Expression
Mol. Pharmacol., August 1, 2007; 72(2): 370 - 379.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Gastrointest. Liver Physiol.Home page
T. A. Markel, P. R. Crisostomo, M. Wang, C. M. Herring, T. Lahm, K. K. Meldrum, K. D. Lillemoe, F. J. Rescorla, and D. R. Meldrum
Iron chelation acutely stimulates fetal human intestinal cell production of IL-6 and VEGF while decreasing HGF: the roles of p38, ERK, and JNK MAPK signaling
Am J Physiol Gastrointest Liver Physiol, April 1, 2007; 292(4): G958 - G963.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
291/2/G260    most recent
00390.2005v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Web of Science (11)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Szalay, L.
Right arrow Articles by Chaudry, I. H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Szalay, L.
Right arrow Articles by Chaudry, I. H.

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Visit Other APS Journals Online
Copyright © 2006 by the American Physiological Society.