Mo1786 HIF-1α Plays a Gut-Injurious Role in Interferon-γ Induced Intestinal Epithelial Barrier...
1
Mo1783 Class III PI3K (HVPS34) Is Involved in AMPK-Regulated Neurotensin Secretion Independent of mTORC1 Signaling Jing Li, Jun Song, Heidi L. Weiss, Tianyan Gao, Courtney M. Townsend, B. Mark Evers Class III PI3K, also known as hVps34, along with its associated regulatory subunit, hVps15 kinase, are critical components of nutrient sensing, and act as upstream factors required for activation of S6K1, a downstream effector of mTOR complex 1 (mTORC1). The hVps34/ hVps15 complex regulates a variety of cellular functions such as intracellular vesicle traffick- ing. In addition, AMPK regulates the hVps34-containing complex in response to changes in energy state. Neurotensin (NT), a gut hormone produced and stored in N cells of the distal small bowel, has multiple physiologic effects, including facilitation of fatty acid translocation from the intestinal lumen, coordination of gut motility and stimulation of pancreaticobiliary secretion. Since we have previously found that AMPK and mTORC1 positively and negatively regulate NT release, respectively, the purpose of the present study was to investigate whether hVps34 and hVps15 are required in AMPK or mTORC1-mediated secretion. METHODS. i) The human endocrine cell line, BON, was used for all experiments. BON cells synthesize and secrete NT peptide and process the NT peptide in a manner analogous to that of N cells in the small bowel, thus serving as a useful model for enteroendo- crine cell secretion. AMPK activation was induced by Aicar or 2-DG, and NT secretion measured using an NT EIA kit. ii) To determine the involvement of hVps34 and hVps15 in AMPK-mediated effects on NT secretion, endogenous expression of hVps34 and hVps15 was inhibited by siRNA-mediated knockdown. Stable BON cell lines overexpressing either the control vector or hVps34/hVps15 (hVps34 and hVps15 are expressed in one vector but regulated by individual promoters) were established. iii) The physical association of hVps34 with AMPK was evaluated by co-immunoprecipitation (co-IP). RESULTS. NT secretion (induced by Aicar or 2-DG) was attenuated in BON cells transfected with siRNAs targeting hVps34 and hVps15 compared to non-targeting control (NTC) siRNA. Overexpression of hVps34/hVps15 increased both basal and Aicar- and 2-DG-stimulated NT secretion. More- over, we detected the physical association of hVps34 with AMPK by co-IP. hVps34 was detected in both AMPKα1- and α2-precipitated complexes with increased hVps34 bound in the AMPKα2 complex. CONCLUSIONS. These results identify a novel regulatory mecha- nism for the hVps34/hVps15 complex in NT peptide secretion, providing a direct connection between AMPK and hVps34 which is independent of mTORC1/S6K1 signaling. Mo1784 Identification of Selective Agonists for the Hpac1 Receptor Samuel A. Mantey, Jerome L. Maderdrut, David Coy, Robert T. Jensen BACKGROUND: Pituitary adenylate cyclase-activating polypeptide (PACAP) and Vasoactive intestinal peptide (VIP) act by 3 classes of G-protein coupled receptors, VPAC1, VPAC2 and PAC1. PAC1 is selective for PACAP, whereas VPAC1 and VPAC2 have high affinity for both VIP and PACAP. PACAP/VIP are widely distributed in the brain and peripheral organs and are thought to be involved in a number of physiological/pathological conditions in the CNS, GI tract and other tissues. Because PACAP interacts with high affinity with all 3 receptor subtypes it is difficult to determine which receptor is responsible for its actions. Furthermore, this is complicated by lack of selective, high affinity PACAP agonists/antago- nists. PURPOSE: The purpose of this study was to attempt to identify analogs of PACAP that were selective for PAC1 over VPAC receptors. METHOD: Using standard solid phase peptide synthesis methodology we initially made 40 peptides with single or multiple amino acid substitutions in PACAP27 and PACAP38 peptides at sites reported to be important for biologic activity, degradation or selectivity for VPAC-PAC receptors. Each of these analogs was tested for their affinities for hVPAC1, hVPAC2 and hPAC1 on native and transfected cells. Biological potencies were studied by assessing increases in [cAMP]i. RESULTS: PACAP27 and PACAP38 had high affinity (IC50: 0.20-3.5 nM) for the 6 cell lines containing PAC1, VPAC1 or VPAC2 receptors. Of the 40 analogs only 6 showed selectivity of at least 5-fold for PAC1. In general, results from native and transfected receptors were similar. An Ala22 substitution resulted in a 5-fold selectivity for PAC1 over VPAC1 and 400-fold over VPAC2. N-terminal acetylation of PACAP resulted in >10-fold greater decrease in affinity for VPAC1 than for VPAC2/PAC1. Similarly D-Ser2-PACAP38 had >30-fold greater decrease in affinity for VPAC1 than for VPAC2/PAC1. From these leads 6 new combination analogs of PACAP38 were made and each showed some selectivity for PAC1. Their selectivity for PAC1 over VPAC2 ranged from 5-330 fold and for VPAC1 from 5-15 fold. The addition of imidazole acetate (Iaa) in position 1 of PACAP alone increased selectivity for the PAC1 6-12 fold. The inclusion of D-Ser2 increased selectivity to >12 fold. The farther inclusion of Ala22 resulted in PACAP38 analogs with >15 fold PAC1 selectivity over VPAC1 and 330 fold over VPAC2. All these analogs were full agonists and changes in binding affinities were also reflected in the biological potentials for activating adenylate cyclase. CONCLUSIONS: This study identifies a number of new PACAP38 analogs that have selectivity for the PACAP receptor, PAC1. These analogs will not only be useful templates for future design for enhancing further selectivity, but will also be useful for investigating the role of the PAC1 in physiological/pathophysiological pro- cesses. Mo1785 Urocortin 3 Expression At Baseline and During Inflammation in the Colon: Corticotropin Releasing Factor Receptors Cross-Talk Shilpi Mahajan, Min Liao, Paris Barkan, Kazuhiro Takahashi, Aditi Bhargava Urocortins (Ucn1-3), members of the corticotropin-releasing factor (CRF) family of neuropep- tides, are emerging as potent immunomodulators. Localized, cellular expression of Ucn1 and Ucn2, but not Ucn3, has been demonstrated during inflammation. In this study, we investigated the expression and the role of Ucn3 in a rat model of Crohn's colitis and the relative contribution of CRF receptors (CRF1 and CRF2) in regulating Ucn3 expression at baseline and during inflammation. Ucn3 was administered in conjunction with or without a bolus of trinitrobenzoic acid enema in the colon to determine its pro or anti-inflammatiory effects. Ucn3 expression was ascertained by RT-PCR and immunohistochemistry. RNAi was S-659 AGA Abstracts used to achieve colon-specific knockdown of CRF1 and CRF2. Ucn3 mRNA and peptide were ubiquitously expressed throughout the GI tract in Naive rats. Ucn3 immunoreactivity was seen in epithelial cells and myenteric neurons. On day 1 of colitis, Ucn3 mRNA levels decreased by 80% and did not recover to baseline even by day 9. Next, we ascertained pro or anti-inflammatory actions of Ucn3 during colitis. Surprisingly, unlike observed anti- inflammatory actions of Ucn1, exogenous Ucn3 did not alter histopathological outcomes during colitis and neither did it alter levels of pro-inflammatory cytokines IL-6 and TNF- α. Phosphorylation of p38 kinase increased by 250% during colitis and was significantly attenuated after Ucn3 administration and knockdown of CRF receptors. At baseline, colon- specific knockdown of CRF1, but not CRF2 decreased Ucn3 mRNA by 78%, whereas during colitis, Ucn3 mRNA levels increased after CRF1 knockdown, suggesting importance of receptor cross-talk. In cultured cells, co-expression of CRF1 + CRF2 attenuated Ucn3- mediated intracellular Ca2+ signaling by 38% as compared to CRF2-mediating Ca2+ signaling by Ucn3. Thus, our results suggest that a balanced and coordinated expression of CRF receptors is required for proper regulation of Ucn3 at baseline and during inflammation. This CRF receptor cross-talk may be important in the design of therapeutic interventions for the CRF system. Mo1786 HIF-1α Plays a Gut-Injurious Role in Interferon-γ Induced Intestinal Epithelial Barrier Dysfunction Songwei Yang, Min Yu, Yingdong Cheng, Lihua Sun, Weidong Xiao, Ligang Sun, Guoqing Chen, Chaojun Zhang, Yuanhang Ma, Hua Yang Background: The proinflammatory cytokine interferon-γ (IFN-γ) plays a vital role in intestinal barrier function disruption, such as inflammatory bowel disease. Hypoxia-inducible factor- 1 (HIF-1) is a critical determinant response to hypoxia and inflammation, which has been shown to be deleterious to intestinal barrier function. In this study, we investigated IFN-γ induces loss of barrier function through regulation of HIF-1α activation and function. Method: T84 intestinal epithelial cells were treated with different doses of IFN-γ for indicated time with or without NF-κB inhibitor (pyrolidinedithiocarbamate,PDTC) and HIF-1α inhibi- tor (3-(5'-Hydroxymethyl-2'-furyl)-1-benzylindazole,YC-1), the expression of HIF-1α and HIF-1β were assayed by western blot. The effect of IFN-γ on NF-κB signaling was assayed by measuring levels of p65 and IκBα phosphorylation/degradation in cytoplasmic and nuclear by Western blot. The intestinal epithelial barrier function was evaluated by transepithelial resistance (TER). The expression and morphological changes of tight junction proteins ZO- 1, Occludin, Claudin-1 and Claudin-2 were observed by immunofluorescence and western blot. Results: IFN-γ led to an increase of HIF-1α expression in time and dose-dependent manners, but did not change the expression of HIF-1β. IFN-γ treatment induced translocation of p65 from the cytoplasm to the nucleus whereas induced a significant increase in cyto- plasmic levels of phosphorylated IκBα and a concomitant decrease in IκBα levels, both of which indicate the activation of NF-κB. The IFN-γ-induced increase in HIF-1α was associated with an activation of NF-κB. Treatment with the NF-κB inhibitor, PDTC, significantly suppressed the activation of NF-κB and the expression of HIF-1α. In addition, IFN-γ also caused time- and dose-dependent drop of TER and depletion of tight junction proteins; inhibition NF-κB (PDTC) or HIF-1α (YC-1) prevented the drop of TER and alteration in tight junction protein expressions and morphological changes. Conclusion: This study suggested that IFN-γ induced the loss of epithelial barrier function and disruption of tight junction proteins, by upregulation of HIF-1α expression through NF-κB pathway. Mo1787 Valproic Acid (VPA), a Histone Deacetylase Inhibitor That Reduces Intraabdominal Adhesions Modulates Peritoneal Plasma Extravasation and Genes That Regulate Fibrin Deposition and Stability Matthew T. Brady, Elizabeth G. King, Michael R. Cassidy, Stanley Heydrick, Arthur F. Stucchi Intraabdominal adhesions occur in nearly 100% of patients following abdominopelvic sur- gery. We have previously shown that VPA administered intraperitoneally (IP) at the time of laparotomy significantly reduces adhesions in a rat model; however, the mechanism remains unclear. We hypothesized that VPA reduces plasma extravasation of the fibrinous exudate at the site of peritoneal injury thus inhibiting fibrin formation in the postoperative period. Twenty-five rats underwent laparotomy with creation of adhesive ischemic buttons as previously described. VPA (50mg/kg) or saline was administered IP intraoperatively followed immediately by a tail vein injection of Evans Blue (25mg/kg). To measure plasma extravasation, animals were sacrificed after 3 hrs and Evans Blue accumulation was quantified in peritoneal tissue. In additional experiments, adhesive buttons were collected 3 hrs postop- eratively for RNA extraction and real-time PCR analysis to quantify mRNA levels of: 1) vascular endothelial growth factor (VEGF; promotes vascular permeability); 2) tissue factor (TF; receptor critical to thrombin formation); and 3) protease-activated-receptor-1 (PAR1; thrombin activated receptor whose signal increases fibrin formation). Four untreated rats served as non-operated controls. The results demonstrate that VPA reduces plasma extravasa- tion into inflamed peritoneal tissue surrounding ischemic buttons. In concert with this effect VPA significantly reduces expression of each of the above genes in adhesive ischemic tissue compared with saline controls (Table). These data mechanistically support that fibrin deposi- tion and stability early in the postoperative period is critical to adhesiogenesis and its regulation is a viable therapeutic approach for adhesion prevention. AGA Abstracts
Transcript of Mo1786 HIF-1α Plays a Gut-Injurious Role in Interferon-γ Induced Intestinal Epithelial Barrier...
Mo1783
Class III PI3K (HVPS34) Is Involved in AMPK-Regulated NeurotensinSecretion Independent of mTORC1 SignalingJing Li, Jun Song, Heidi L. Weiss, Tianyan Gao, Courtney M. Townsend, B. Mark Evers
Class III PI3K, also known as hVps34, along with its associated regulatory subunit, hVps15kinase, are critical components of nutrient sensing, and act as upstream factors required foractivation of S6K1, a downstream effector of mTOR complex 1 (mTORC1). The hVps34/hVps15 complex regulates a variety of cellular functions such as intracellular vesicle traffick-ing. In addition, AMPK regulates the hVps34-containing complex in response to changesin energy state. Neurotensin (NT), a gut hormone produced and stored in N cells ofthe distal small bowel, has multiple physiologic effects, including facilitation of fatty acidtranslocation from the intestinal lumen, coordination of gut motility and stimulation ofpancreaticobiliary secretion. Since we have previously found that AMPK and mTORC1positively and negatively regulate NT release, respectively, the purpose of the present studywas to investigate whether hVps34 and hVps15 are required in AMPK or mTORC1-mediatedsecretion. METHODS. i) The human endocrine cell line, BON, was used for all experiments.BON cells synthesize and secrete NT peptide and process the NT peptide in a manneranalogous to that of N cells in the small bowel, thus serving as a useful model for enteroendo-crine cell secretion. AMPK activation was induced by Aicar or 2-DG, and NT secretionmeasured using an NT EIA kit. ii) To determine the involvement of hVps34 and hVps15in AMPK-mediated effects on NT secretion, endogenous expression of hVps34 and hVps15was inhibited by siRNA-mediated knockdown. Stable BON cell lines overexpressing eitherthe control vector or hVps34/hVps15 (hVps34 and hVps15 are expressed in one vector butregulated by individual promoters) were established. iii) The physical association of hVps34with AMPK was evaluated by co-immunoprecipitation (co-IP). RESULTS. NT secretion(induced by Aicar or 2-DG) was attenuated in BON cells transfected with siRNAs targetinghVps34 and hVps15 compared to non-targeting control (NTC) siRNA. Overexpression ofhVps34/hVps15 increased both basal and Aicar- and 2-DG-stimulated NT secretion. More-over, we detected the physical association of hVps34 with AMPK by co-IP. hVps34 wasdetected in both AMPKα1- and α2-precipitated complexes with increased hVps34 boundin the AMPKα2 complex. CONCLUSIONS. These results identify a novel regulatory mecha-nism for the hVps34/hVps15 complex in NT peptide secretion, providing a direct connectionbetween AMPK and hVps34 which is independent of mTORC1/S6K1 signaling.
Mo1784
Identification of Selective Agonists for the Hpac1 ReceptorSamuel A. Mantey, Jerome L. Maderdrut, David Coy, Robert T. Jensen
BACKGROUND: Pituitary adenylate cyclase-activating polypeptide (PACAP) and Vasoactiveintestinal peptide (VIP) act by 3 classes of G-protein coupled receptors, VPAC1, VPAC2and PAC1. PAC1 is selective for PACAP, whereas VPAC1 and VPAC2 have high affinity forboth VIP and PACAP. PACAP/VIP are widely distributed in the brain and peripheral organsand are thought to be involved in a number of physiological/pathological conditions in theCNS, GI tract and other tissues. Because PACAP interacts with high affinity with all 3receptor subtypes it is difficult to determine which receptor is responsible for its actions.Furthermore, this is complicated by lack of selective, high affinity PACAP agonists/antago-nists. PURPOSE: The purpose of this study was to attempt to identify analogs of PACAPthat were selective for PAC1 over VPAC receptors. METHOD: Using standard solid phasepeptide synthesis methodology we initially made 40 peptides with single or multiple aminoacid substitutions in PACAP27 and PACAP38 peptides at sites reported to be important forbiologic activity, degradation or selectivity for VPAC-PAC receptors. Each of these analogswas tested for their affinities for hVPAC1, hVPAC2 and hPAC1 on native and transfected cells.Biological potencies were studied by assessing increases in [cAMP]i. RESULTS: PACAP27 andPACAP38 had high affinity (IC50: 0.20-3.5 nM) for the 6 cell lines containing PAC1, VPAC1or VPAC2 receptors. Of the 40 analogs only 6 showed selectivity of at least 5-fold for PAC1.In general, results from native and transfected receptors were similar. An Ala22 substitutionresulted in a 5-fold selectivity for PAC1 over VPAC1 and 400-fold over VPAC2. N-terminalacetylation of PACAP resulted in >10-fold greater decrease in affinity for VPAC1 than forVPAC2/PAC1. Similarly D-Ser2-PACAP38 had >30-fold greater decrease in affinity for VPAC1than for VPAC2/PAC1. From these leads 6 new combination analogs of PACAP38 weremade and each showed some selectivity for PAC1. Their selectivity for PAC1 over VPAC2ranged from 5-330 fold and for VPAC1 from 5-15 fold. The addition of imidazole acetate(Iaa) in position 1 of PACAP alone increased selectivity for the PAC1 6-12 fold. The inclusionof D-Ser2 increased selectivity to >12 fold. The farther inclusion of Ala22 resulted in PACAP38analogs with >15 fold PAC1 selectivity over VPAC1 and 330 fold over VPAC2. All theseanalogs were full agonists and changes in binding affinities were also reflected in the biologicalpotentials for activating adenylate cyclase. CONCLUSIONS: This study identifies a numberof new PACAP38 analogs that have selectivity for the PACAP receptor, PAC1. These analogswill not only be useful templates for future design for enhancing further selectivity, but willalso be useful for investigating the role of the PAC1 in physiological/pathophysiological pro-cesses.
Mo1785
Urocortin 3 Expression At Baseline and During Inflammation in the Colon:Corticotropin Releasing Factor Receptors Cross-TalkShilpi Mahajan, Min Liao, Paris Barkan, Kazuhiro Takahashi, Aditi Bhargava
Urocortins (Ucn1-3), members of the corticotropin-releasing factor (CRF) family of neuropep-tides, are emerging as potent immunomodulators. Localized, cellular expression of Ucn1and Ucn2, but not Ucn3, has been demonstrated during inflammation. In this study, weinvestigated the expression and the role of Ucn3 in a rat model of Crohn's colitis and therelative contribution of CRF receptors (CRF1 and CRF2) in regulating Ucn3 expression atbaseline and during inflammation. Ucn3 was administered in conjunction with or withouta bolus of trinitrobenzoic acid enema in the colon to determine its pro or anti-inflammatioryeffects. Ucn3 expression was ascertained by RT-PCR and immunohistochemistry. RNAi was
S-659 AGA Abstracts
used to achieve colon-specific knockdown of CRF1 and CRF2. Ucn3 mRNA and peptidewere ubiquitously expressed throughout the GI tract in Naive rats. Ucn3 immunoreactivitywas seen in epithelial cells and myenteric neurons. On day 1 of colitis, Ucn3 mRNA levelsdecreased by 80% and did not recover to baseline even by day 9. Next, we ascertained proor anti-inflammatory actions of Ucn3 during colitis. Surprisingly, unlike observed anti-inflammatory actions of Ucn1, exogenous Ucn3 did not alter histopathological outcomesduring colitis and neither did it alter levels of pro-inflammatory cytokines IL-6 and TNF-α. Phosphorylation of p38 kinase increased by 250% during colitis and was significantlyattenuated after Ucn3 administration and knockdown of CRF receptors. At baseline, colon-specific knockdown of CRF1, but not CRF2 decreased Ucn3 mRNA by 78%, whereas duringcolitis, Ucn3 mRNA levels increased after CRF1 knockdown, suggesting importance ofreceptor cross-talk. In cultured cells, co-expression of CRF1 + CRF2 attenuated Ucn3-mediated intracellular Ca2+ signaling by 38% as compared to CRF2-mediating Ca2+ signalingby Ucn3. Thus, our results suggest that a balanced and coordinated expression of CRFreceptors is required for proper regulation of Ucn3 at baseline and during inflammation.This CRF receptor cross-talk may be important in the design of therapeutic interventionsfor the CRF system.
Mo1786
HIF-1α Plays a Gut-Injurious Role in Interferon-γ Induced Intestinal EpithelialBarrier DysfunctionSongwei Yang, Min Yu, Yingdong Cheng, Lihua Sun, Weidong Xiao, Ligang Sun, GuoqingChen, Chaojun Zhang, Yuanhang Ma, Hua Yang
Background: The proinflammatory cytokine interferon-γ (IFN-γ) plays a vital role in intestinalbarrier function disruption, such as inflammatory bowel disease. Hypoxia-inducible factor-1 (HIF-1) is a critical determinant response to hypoxia and inflammation, which has beenshown to be deleterious to intestinal barrier function. In this study, we investigated IFN-γinduces loss of barrier function through regulation of HIF-1α activation and function.Method: T84 intestinal epithelial cells were treated with different doses of IFN-γ for indicatedtime with or without NF-κB inhibitor (pyrolidinedithiocarbamate,PDTC) and HIF-1α inhibi-tor (3-(5'-Hydroxymethyl-2'-furyl)-1-benzylindazole,YC-1), the expression of HIF-1α andHIF-1β were assayed by western blot. The effect of IFN-γ on NF-κB signaling was assayedby measuring levels of p65 and IκBα phosphorylation/degradation in cytoplasmic and nuclearby Western blot. The intestinal epithelial barrier function was evaluated by transepithelialresistance (TER). The expression and morphological changes of tight junction proteins ZO-1, Occludin, Claudin-1 and Claudin-2 were observed by immunofluorescence and westernblot. Results: IFN-γ led to an increase of HIF-1α expression in time and dose-dependentmanners, but did not change the expression of HIF-1β. IFN-γ treatment induced translocationof p65 from the cytoplasm to the nucleus whereas induced a significant increase in cyto-plasmic levels of phosphorylated IκBα and a concomitant decrease in IκBα levels, both ofwhich indicate the activation of NF-κB. The IFN-γ-induced increase in HIF-1α was associatedwith an activation of NF-κB. Treatment with the NF-κB inhibitor, PDTC, significantlysuppressed the activation of NF-κB and the expression of HIF-1α. In addition, IFN-γ alsocaused time- and dose-dependent drop of TER and depletion of tight junction proteins;inhibition NF-κB (PDTC) or HIF-1α (YC-1) prevented the drop of TER and alterationin tight junction protein expressions and morphological changes. Conclusion: This studysuggested that IFN-γ induced the loss of epithelial barrier function and disruption of tightjunction proteins, by upregulation of HIF-1α expression through NF-κB pathway.
Mo1787
Valproic Acid (VPA), a Histone Deacetylase Inhibitor That ReducesIntraabdominal Adhesions Modulates Peritoneal Plasma Extravasation andGenes That Regulate Fibrin Deposition and StabilityMatthew T. Brady, Elizabeth G. King, Michael R. Cassidy, Stanley Heydrick, Arthur F.Stucchi
Intraabdominal adhesions occur in nearly 100% of patients following abdominopelvic sur-gery. We have previously shown that VPA administered intraperitoneally (IP) at the timeof laparotomy significantly reduces adhesions in a rat model; however, the mechanismremains unclear. We hypothesized that VPA reduces plasma extravasation of the fibrinousexudate at the site of peritoneal injury thus inhibiting fibrin formation in the postoperativeperiod. Twenty-five rats underwent laparotomy with creation of adhesive ischemic buttonsas previously described. VPA (50mg/kg) or saline was administered IP intraoperativelyfollowed immediately by a tail vein injection of Evans Blue (25mg/kg). To measure plasmaextravasation, animals were sacrificed after 3 hrs and Evans Blue accumulation was quantifiedin peritoneal tissue. In additional experiments, adhesive buttons were collected 3 hrs postop-eratively for RNA extraction and real-time PCR analysis to quantify mRNA levels of: 1)vascular endothelial growth factor (VEGF; promotes vascular permeability); 2) tissue factor(TF; receptor critical to thrombin formation); and 3) protease-activated-receptor-1 (PAR1;thrombin activated receptor whose signal increases fibrin formation). Four untreated ratsserved as non-operated controls. The results demonstrate that VPA reduces plasma extravasa-tion into inflamed peritoneal tissue surrounding ischemic buttons. In concert with this effectVPA significantly reduces expression of each of the above genes in adhesive ischemic tissuecompared with saline controls (Table). These data mechanistically support that fibrin deposi-tion and stability early in the postoperative period is critical to adhesiogenesis and itsregulation is a viable therapeutic approach for adhesion prevention.
AG
AA
bst
ract
s
