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1.
J Huazhong Univ Sci Technolog Med Sci ; 29(4): 486-91, 2009 Aug.
Article in English | MEDLINE | ID: mdl-19662368

ABSTRACT

The purpose of this study was to investigate bacterial translocation and change in intestinal permeability in patients after abdominal surgery. Sixty-three patients undergoing elective abdominal surgery were enrolled in the study. Blood samples were collected prior to operation and 2, 24, 48 h after surgery for bacterial culture, microbial DNA extraction, plasma D-lactate and endotoxin measurement. PCR analysis was performed after DNA extraction, with beta-lactosidase gene of E. coli and 16S rRNA gene as target genes. All patients were observed for a period of 30 days for infectious complications. Our results showed that no bacterial DNA was detected before surgery, but after operation it was found in 12 patients (19.0%). Bacterial DNA was detected in 41.7% (10/24) of SIRS patients and 5.1% (2/39) of non-SIRS patients (P<0.01). About 83.3% of PCR-positive patients developed systemic inflammatory response syndrome (SIRS), but only 27.5% of PCR-negative patients did so (P<0.01). Two thirds of PCR-positive patients developed infectious complications, while none of PCR-negative patients did (P<0.01). The blood culture was positive only in 3 patients (4.8%), who were all PCR-positive. E. coli DNA was found in 66.7% of the PCR-positive patients. The plasma levels of D-lactate and endotoxin were elevated significantly 2, 24 and 48 h after operation in PCR-positive patients, with a significant positive correlation found between them (r=0.91, P<0.01). It is concluded that increased intestinal permeability was closely related with bacterial translocation. Intestinal bacterial translocation (most commonly E. coli) might occur at early stage (2 h) after abdominal surgery. Postoperative SIRS and infection might bear a close relationship with bacterial translocation.


Subject(s)
Bacterial Translocation/physiology , Cell Membrane Permeability/physiology , Escherichia coli/physiology , Intestinal Mucosa/physiopathology , Postoperative Complications , Adult , Aged , Cholecystectomy , DNA, Bacterial/metabolism , Female , Gastrectomy , Humans , Male , Middle Aged
2.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-301287

ABSTRACT

sely related with bacterial translocation. Intestinal bacterial translocation (most commonly E. coli) might occur at early stage (2 h) after ab-dominal surgery. Postoperative SIRS and infection might bear a close relationship with bacterial translocation.

3.
Article in Chinese | MEDLINE | ID: mdl-16464377

ABSTRACT

OBJECTIVE: To investigate bacterial translocation in severe multiple trauma patients using polymerase chain reaction (PCR) to detect the presence of bacteria in the blood. METHODS: Sixteen severe multiple trauma patients [injury severity score (ISS)>20] in surgery intensive care unit (SICU) were selected. Blood samples were collected 2, 24 and 48 hours after trauma for bacterial culture and microbial DNA detection. Meanwhile, plasma levels of D-lactate and lipopolysaccharide (LPS) in systemic circulation were determined. PCR was performed after DNA extraction, with target beta-lactosidase gene of E. coli and 16SrRNA gene of most pathogenic bacteria. All patients were observed within 30 days for infectious complications. D-lactate and LPS levels were determined in 63 patients before selective operation. RESULTS: Microbial DNA could be detected in blood as early as 2 hours following severe trauma, and altogether positive results were found in 10 patients (62.50%). All PCR-positive patients manifested sepsis, but none of the PCR-negative patients did (P<0.01). Bacterial DNA was discovered in 100.00% of sepsis patients and none in non-sepsis patients (P<0.01). Seventy percent of PCR-positive patients developed infectious complications, while none of PCR-negative patients did (P<0.01). The blood culture was positive only in 3 patients (18.75%), all of them were PCR-positive. E.coli DNA was found in 70.00% of all the PCR positive blood specimens. Systemic plasma concentration of D-lactate and LPS of all patients was significantly higher than that in control group, which consisted of 63 inpatients waiting for elective operations. Systemic plasma level of D-lactate showed a positive correlation with that of LPS (r=0.94, P<0.01). CONCLUSION: Intestinal bacterial translocation (most commonly E. coli) might occur early (2 hours) after severe trauma. Infection and sepsis have a close relationship with bacterial translocation. Detection of blood microbial DNA using PCR could reflect bacteria translocation and forecast imminent infection and sepsis.


Subject(s)
Bacterial Translocation , Intestines/microbiology , Multiple Trauma/microbiology , Adolescent , Adult , DNA, Bacterial/blood , DNA, Bacterial/isolation & purification , Female , Humans , Lactic Acid/blood , Lipopolysaccharides/blood , Male , Middle Aged , Multiple Trauma/complications , Sepsis/etiology , Young Adult
4.
Burns ; 30(3): 225-31, 2004 May.
Article in English | MEDLINE | ID: mdl-15082348

ABSTRACT

Recent studies have suggested that levels of lipopolysaccharide-binding protein (LBP) might play a harmful role by up-regulating the host's sensitivity to endotoxin. Our previous studies demonstrated that local endotoxin could up-regulate LBP expression after acute insults, however, the definite molecular mechanisms downstream of endotoxin action remain unclear. This study investigates whether tumor necrosis factor (TNF-alpha) might be responsible for the LBP formation during endogenous endotoxemia postburn. Wistar rats were anesthetized, and a 35% TBSA full-thickness burn was created. Animals were randomly divided into normal control, thermal injury and anti-TNF-alpha mAb treatment group. A significant elevation of plasma endotoxin concentration was observed after acute insults. TNF-alpha levels in plasma also rapidly increased after thermal injury. Meanwhile, LBP mRNA expression markedly increased in liver, lungs, kidneys and intestine postburn. There was no detectable TNF-alpha in the plasma of anti-TNF-alpha mAb treated animals. Treatment with anti-TNF-alpha mAb also resulted in significantly lower concentrations of LBP mRNA in local tissues. Additionally, several organ function parameter levels in plasma significantly decreased in treatment group. These results demonstrated that an increase of plasma TNF-alpha levels caused by burns might be associated with a marked elevation of tissue LBP mRNA expression, which could contribute to the development of multiple organ damage.


Subject(s)
Acute-Phase Proteins , Burns/metabolism , Carrier Proteins/metabolism , Endotoxins/metabolism , Membrane Glycoproteins , Tumor Necrosis Factor-alpha/physiology , Animals , Male , RNA, Messenger/metabolism , Random Allocation , Rats , Rats, Wistar , Reverse Transcriptase Polymerase Chain Reaction/methods , Up-Regulation
5.
Zhonghua Wai Ke Za Zhi ; 41(8): 620-2, 2003 Aug.
Article in Chinese | MEDLINE | ID: mdl-14505541

ABSTRACT

OBJECTIVE: To explore if early fracture fixation can alleviate gut barrier function damage caused by multiple firearm injuries in pigs. METHODS: Twelve healthy pigs were subjected to tangential fracture of parietal bone and comminuted fractures of bilateral femora (ISS >or= 16) due to 5.8 mm bullets shooting and these pigs were divided randomly into 2 groups. Control group (n = 6) were not treated at all. Fracture fixation Group (n = 6) were managed by immediate fracture fixation of bilateral femora with intramedullary nails. Plasma concentration of D-lactate, DAO and endotoxin (in portal vein) were detected at different intervals before and after trauma. The portal vein blood was cultured and the percentage of positive isolation was calculated. The concentration of DAO in small bowel was also detected 72 hours later after trauma. RESULTS: In control group, the plasma concentrations of D-lactate, DAO and endotoxin increased at early stage and kept high till 72 hours after trauma; the percentage of positive blood culture was 63.3%. In Group F, the levels of plasma D-lactate, DAO and endotoxin were also elevated at early stage (6 - 12 h), but declined significantly from 24 h or 48 h after trauma compared with control group (P < 0.05), and the percentage of positive blood culture was lower (30.0%, P < 0.05). The concentrations of DAO in small bowel decreased in both groups, but to a less extent in Group F. CONCLUSION: Bacterial and endotoxin translocation emerged with increasing gut permeability after multiple firearm injuries. The damage of gut barrier function could be alleviated and the chance of enterogenous infection could be by early fracture fixation after trauma.


Subject(s)
Fracture Fixation, Internal , Multiple Trauma/surgery , Wounds, Gunshot/surgery , Animals , Disease Models, Animal , Intestinal Mucosa/metabolism , Multiple Trauma/physiopathology , Permeability , Random Allocation , Swine , Wounds, Gunshot/physiopathology
6.
Shock ; 20(3): 257-63, 2003 Sep.
Article in English | MEDLINE | ID: mdl-12923498

ABSTRACT

Staphylococcal enterotoxin B (SEB) is an important member of the superantigen family, which exerts a number of pathological effects in the human, as well as susceptible animals. The present study was conducted to observe the time course and tissue distribution of SEB in postburn Staphylococcus aureus infection; meanwhile, the relationship between SEB and multiple organ dysfunction was also studied. Eighty-six male Wistar rats were randomly divided into four groups as follows: normal control group (n = 10); scald control group (n = 10); postburn sepsis group (n = 50) in which rats inflicted with 20% total body surface area (TBSA) III degrees scald followed by SEB-producing S. aureus challenge were further divided into 0.5-, 2-, 6-, 12-, and 24-h subgroups, with 10 rats in each subgroup; and SEB monoclonal antibody (MAb) treatment group (n = 16) in which a dose of 4 mg/kg SEB MAb was given intravenously just before S. aureus challenge, and the rats were further divided into 2- and 6-h subgroups. It was found that after thermal injury combined with S. aureus infection, SEB was widely distributed to the liver, kidneys, lungs, and heart, exacerbating the pathophysiology of multiple organ dysfunction induced by postburn sepsis. At the same time, the gene and protein expressions of tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) were also markedly upregulated in various tissues. Early treatment with SEB-specific MAb-MAb2D(1)-could markedly decrease SEB levels in plasma as well as in various tissues, and could significantly reduce the 6-h mortality rate (17.64% [3/17] vs. 55.6% [20/36], P = 0.02). These data suggested that neutralization of SEB is effective in ameliorating S. aureus sepsis and subsequent multiple organ damage, which might be attributed to its inhibitory effect on inflammatory mediator formation.


Subject(s)
Enterotoxins/metabolism , Sepsis/microbiology , Sepsis/pathology , Staphylococcus aureus/pathogenicity , Animals , Antibodies, Monoclonal/metabolism , Burns , Enterotoxins/blood , Interferon-gamma/biosynthesis , Male , Rats , Rats, Wistar , Staphylococcal Infections/pathology , Time Factors , Tissue Distribution , Tumor Necrosis Factor-alpha/biosynthesis
7.
Shock ; 20(2): 159-65, 2003 Aug.
Article in English | MEDLINE | ID: mdl-12865661

ABSTRACT

It has been demonstrated that biopterin, an essential cofactor of nitric oxide synthase (NOS), plays an important role in the pathogenesis of endotoxin-induced shock, yet its biological significance in gram-positive sepsis remains unclear. In this study, we adopted a rat model of postburn Staphylococcus aureus sepsis to investigate the potential role of biopterin in the pathogenesis of gram-positive sepsis. Wistar rats were inflicted with a 20% total body surface area (TBSA) full-thickness scald injury followed by S. aureus challenge, and then guanosine triphosphate-cyclohydrolase I (GTP-CHI) mRNA expression and biopterin levels in liver, kidneys, lungs, and heart were determined. We found that after S. aureus challenge, GTP-CHI gene expressions and biopterin levels were markedly upregulated in various tissues. Meanwhile, multiple organ dysfunction was induced by S. aureus challenge. It was shown that cardiac GTP-CHI mRNA expression and renal BH(4) levels were positively correlated with MB isoenzyme of creatine kinase (CK-MB) and creatinine (r = 0.892, P = 0.0012 and r = 0.9423, P = 0.0015, respectively). These results suggested that thermal injury combined with S. aureus challenge could induce de novo biosynthesis of biopterin, which might play a role in the development of multiple organ dysfunction syndrome secondary to postburn sepsis.


Subject(s)
Biopterins/biosynthesis , Burns/complications , Sepsis/metabolism , Staphylococcal Infections/metabolism , Staphylococcus aureus/pathogenicity , Animals , Creatine Kinase/chemistry , Creatinine/chemistry , Male , Nitric Oxide/blood , Nitric Oxide Synthase/metabolism , Nitric Oxide Synthase Type II , Protein Isoforms , RNA, Messenger/metabolism , Rats , Rats, Wistar , Shock , Time Factors , Tissue Distribution
8.
World J Gastroenterol ; 9(5): 1038-44, 2003 May.
Article in English | MEDLINE | ID: mdl-12717852

ABSTRACT

AIM: To investigate changes of tumor necrosis factor-alpha (TNF-alpha) and TNFR-I expression in vital organs and their significance in the pathogenesis of multiple organ damage associated with endogenous endotoxin following major burns. METHODS: Wistar rats subjected to a 35 % full-thickness scald injury were sacrificed at 12 h, 24 h, 48 h, and 72 h postburn, respectively. Meanwhile, eight rats were taken as normal controls. Tissue samples from liver, spleen, kidney, lung and intestine were collected to assay tissue endotoxin levels and measure TNF-alpha and TNFR-I expression. In addition, blood samples were obtained for the determination of organ function parameters. RESULTS: Endotoxin levels in liver, spleen and lung increased markedly after thermal injury, with the highest level in liver. The gene expression of TNF-alpha in liver, lung and kidney was up-regulated after thermal injury, while the TNFR-I mRNA expression in liver, lung, kidney and intestine was shown decreased throughout the observation period. Thus, the mRNA expression ratio of TNF-alpha to TNFR-I was significantly increased postburn, particularly in pulmonary tissue (67-fold). In addition, the significant correlations between the expression of TNFR-I or the expression ratio of TNF-alpha/TNFR mRNA in liver tissue and serum aspartate aminotransferase levels were noted (P<0.05-0.01). Similar results were also obtained between pulmonary TNF-alpha mRNA expression and myeloperoxidase activities (P<0.01), whereas there was a highly negative correlation between levels of renal TNFR-I mRNA expression and serum creatinine. CONCLUSION: Burn injury could result in the translocation of gut-derived endotoxin that was mainly distributed in the liver, spleen and lung. The translocated endotoxin then made the expression of TNF-alpha and TNFR-I mRNA up-regulated and down-regulated respectively in various organs, which might be involved in the pathogenesis of multiple organ damage following burns.


Subject(s)
Antigens, CD/genetics , Burns/genetics , RNA, Messenger/genetics , RNA, Messenger/metabolism , Receptors, Tumor Necrosis Factor/genetics , Tumor Necrosis Factor-alpha/genetics , Animals , Base Sequence , Burns/complications , Burns/microbiology , Burns/physiopathology , DNA/genetics , Endotoxins/metabolism , Gene Expression , Gram-Negative Bacteria/isolation & purification , Gram-Negative Bacteria/pathogenicity , Gram-Negative Bacterial Infections/etiology , Gram-Negative Bacterial Infections/physiopathology , Intestines/physiopathology , Kidney/physiopathology , Liver/physiopathology , Lung/physiopathology , Male , Multiple Organ Failure/etiology , Multiple Organ Failure/genetics , Multiple Organ Failure/microbiology , Multiple Organ Failure/physiopathology , Rats , Rats, Wistar , Receptors, Tumor Necrosis Factor, Type I , Sepsis/etiology , Sepsis/physiopathology , Tissue Distribution
9.
Zhonghua Jie He He Hu Xi Za Zhi ; 25(7): 403-7, 2002 Jul.
Article in English | MEDLINE | ID: mdl-12435298

ABSTRACT

OBJECTIVE: To study the nature of extracellular matirx (ECM) remodeling and its role in airflow obstruction in a rat model of chronic obstructive pulmonary disease (COPD), and to observe the role of nacetylcystein (NAC), protein kinase C (H(7)) and TGF-beta monocolonal antibody in the regulation of extracellular matrix remodeling in the airway wall. METHODS: Fifty-three Wistar rats were randomly divided into 5 groups: the healthy control group, the COPD model group, the NAC group, the H(7) group and the TGF-beta monocolonal antibody group. Pathologic study of the airway and lung tissue, lung function test and blood gas analysis were performed. Fibroblasts and lymphocytes of the bronchial wall and alveolar macrophages were counted. Areas of the epithelial layer, the smooth muscle layer and the lamina propria were measured by image analyzer. The level of hydroxyproline in bronchial and lung homogenates was determined by biochemistry method. The serum levels of laminin (LN) and hyaluronic acid (HA) were determined by RIA method. RESULTS: The changes in histopathology, lung function and blood gas in the animal model were similar to those in COPD patients. The collagen, mainly type I collagen, in airway walls was significantly increased. The areas of the epithelial layer (21 114 micro m(2)) and the smooth muscle layer (16 061 micro m(2)) were significantly increased in the COPD model as compared to the control group (13 056 micro m(2) and 6 692 micro m(2), respectively) (P < 0.01). In the drug intervention groups these parameters were significantly decreased compared to the control group. The numbers of fibroblasts (13.6 +/- 4.2), lymphocytes (35.6 +/- 6.4) and alveolar macrophages (14.8 +/- 1.1) in the model group, were significantly increased compared to the control group (6.8 +/- 1.4, 6.1 +/- 1.2 and 3.5 +/- 1.2, respectively) (P < 0.01, 0.001, 0.001), while in the drug intervention groups the cells were significantly decreased except for fibroblasts in the H(7) group. The hydroxyproline level of the model group (111.5 +/- 2.3) pg/ml was significantly increased as compared to the control group (47.8 +/- 9.7) pg/ml (P < 0.05) and was negatively correlated with FEV(0.3)/FVC (P < 0.001) and positively correlated with airflow resistance (P < 0.01). The number of fibroblasts was also positively correlated with the level of hydroxyproline (P < 0.001). The serum levels of LN (26 +/- 4) micro m/L and HA (19.4 +/- 1.4) micro g/L in the model group were significantly increased compared to the control group (15 +/- 3) micro g/L, and (10.9 +/- 2.9) micro g/L, respectively (P < 0.05). Hydroxyproline in the NAC group (83.1 +/- 41.7) pg/ml and the TGF-beta monoclonal antibody group (71.2 +/- 20.3) pg/ml was significantly decreased, while in the H(7) group (160.6 +/- 41.7) pg/ml it was significantly increased. CONCLUSION: Excessive deposition of ECM, mainly of type I collagen, and proliferation of functionally activated fibroblasts were important pathological changes in airway remodeling and the important causes of airflow obstruction. TGF-beta monoclonal antibody and NAC can modulate airway extracellular matrix remodeling. H(7) can increase collagen deposition in the airway wall but the underlining mechanisms need to be elucidated.


Subject(s)
Lung , Pulmonary Disease, Chronic Obstructive , Animals , Disease Models, Animal , Extracellular Matrix/metabolism , Humans , Rats , Rats, Wistar , Transforming Growth Factor beta/metabolism
10.
J Trauma ; 53(5): 957-67, 2002 Nov.
Article in English | MEDLINE | ID: mdl-12435950

ABSTRACT

BACKGROUND: We hypothesized that lipopolysaccharide-binding protein (LBP) and lipopolysaccharide receptor CD14 would present a pair of key molecules in pathophysiologic alterations induced by low concentrations of endotoxin after trauma. The aim of this study was to investigate the relationship between endotoxin translocation and tissue LBP/CD14 messenger ribonucleic acid (mRNA) expression after burn injury, and to define the potential role of LBP/CD14 in mediating inflammatory mediator induction, as well as the pathogenesis of organ damage. METHODS: Wistar rats were subjected to a 35% full-thickness scald injury, and tissue samples from liver, kidneys, lungs, and intestine were collected to measure LBP/CD14 and tumor necrosis factor-alpha (TNF-alpha) mRNA expression. Peritoneal macrophages were harvested by peritoneal lavage to determine CD14 mRNA expression. RESULTS: It was found that endotoxin levels in liver, spleen, and lung increased markedly after thermal injury, with the highest level in liver. Both tissue LBP and CD14 mRNA expression increased markedly after burns, peaking at 12 hours, and then decreasing gradually. At 48 hours, LBP gene expression had a tendency to the baseline level, whereas CD14 mRNA expression increased again. Likewise, CD14 mRNA levels were up-regulated markedly in peritoneal macrophages. Conversely, gene expression of TNF-alpha in tissues elevated markedly after acute insults. There were positive correlations between lipopolysaccharide levels and LBP/CD14 mRNA as well as TNF-alpha mRNA expression in tissues. Similar results were also obtained between CD14, TNF-alpha mRNA expression in liver tissue and liver function parameters, and between pulmonary TNF-alpha mRNA and myeloperoxidase activities (p < 0.01). CONCLUSION: Thermal injury per se can markedly up-regulate both LBP and CD14 gene expression in various organs. Excessive LBP and CD14 mRNA expression might be associated with enhanced synthesis and release of TNF-alpha stimulated by endotoxin translocation after major burns.


Subject(s)
Acute-Phase Proteins , Burns/metabolism , Carrier Proteins/genetics , Gene Expression , Lipopolysaccharide Receptors/genetics , Membrane Glycoproteins , RNA, Messenger/analysis , Amine Oxidase (Copper-Containing)/metabolism , Animals , Burns/physiopathology , Endotoxins/metabolism , In Situ Hybridization , Male , Peroxidase/metabolism , Rats , Rats, Wistar , Reverse Transcriptase Polymerase Chain Reaction , Statistics, Nonparametric , Tumor Necrosis Factor-alpha/genetics , Up-Regulation
11.
Crit Care Med ; 30(11): 2520-7, 2002 Nov.
Article in English | MEDLINE | ID: mdl-12441764

ABSTRACT

OBJECTIVE: Guanosine triphosphate-cyclohydrolase I (GTP-CHI) is the first and rate-limiting enzyme for the de novo biosynthesis of biopterin. The objective of present study was to observe the effect of 2,4-diamino-6-hydroxy-pyrimidine (DAHP), an inhibitor of GTP-CHI, on the development of postburn Staphylococcus aureus sepsis. DESIGN: A prospective, controlled animal study. SETTING: A research laboratory in a hospital. SUBJECTS: Male Wistar rats. INTERVENTIONS: Fifty-six male Wistar rats were randomly divided into four groups as follows: normal control group (n = 10), scald control group (n = 10), postburn sepsis group (n = 20), and DAHP treatment group (n = 16). In the scald control group, rats were subjected to a 20% total body surface area third-degree scald injury and then were killed at 24 hrs. In the postburn sepsis group (n = 20), rats were inflicted with 20% total body surface area third-degree scald followed by Staphylococcus aureus challenge, and they were further divided into 2- and 6-hr groups. In the DAHP treatment group (n = 16), animals were intraperitoneally injected with a dose of 1 g/kg DAHP before Staphylococcus aureus challenge and then were further divided into 2- and 6-hr groups. Tissue samples from liver, kidneys, lungs, and heart were collected to determine GTP-CHI, inducible nitric oxide synthase, and tumor necrosis factor-alpha messenger RNA expression. Meanwhile, biopterin and nitric oxide concentrations in these tissues were also measured. MEASUREMENTS AND MAIN RESULTS: After the scald injury followed by Staphylococcus aureus challenge, GTP-CHI messenger RNA expression and biopterin concentrations were significantly elevated in various tissues such as liver, heart, kidneys, and lungs, as were the values of inducible nitric oxide synthase messenger RNA expression and nitric oxide formation (p <.01). Pretreatment with DAHP significantly reduced GTP-CHI/biopterin induction (p <.05-.01), and the up-regulation of inducible nitric oxide synthase/nitric oxide was also suppressed. Furthermore, DAHP administration inhibited the gene expression of tumor necrosis factor-alpha. Two hours after septic challenge, tumor necrosis factor-alpha messenger RNA expression in liver, kidneys, and lungs in the DAHP-treated group was 35.7%, 37.3%, and 33.0% of that in the postburn septic group, respectively. Additionally, in animals without DAHP treatment, the 6-hr mortality rate was 55.6% (20 of 36), whereas it was only 25.0% in DAHP-treated animals (4 of 16, p =.08). CONCLUSIONS: Early treatment with DAHP might be a potential strategy to prevent the development of postburn Staphylococcal sepsis, which appears to be associated with down-regulation of biopterin and nitric oxide formation by DAHP.


Subject(s)
Biopterins/analogs & derivatives , Burns/microbiology , Enzyme Inhibitors/therapeutic use , GTP Cyclohydrolase/antagonists & inhibitors , Hypoxanthines/therapeutic use , Sepsis/prevention & control , Staphylococcal Infections/prevention & control , Analysis of Variance , Animals , Biopterins/biosynthesis , Down-Regulation/drug effects , Enzyme Inhibitors/pharmacology , Hypoxanthines/pharmacology , Male , Nitric Oxide/biosynthesis , Nitric Oxide Synthase/biosynthesis , Prospective Studies , RNA, Messenger/metabolism , Random Allocation , Rats , Rats, Wistar , Sepsis/etiology , Sepsis/microbiology , Staphylococcal Infections/etiology
12.
Zhonghua Nei Ke Za Zhi ; 41(6): 393-8, 2002 Jun.
Article in Chinese | MEDLINE | ID: mdl-12137602

ABSTRACT

OBJECTIVE: To study the role of metalloproteinases(MMPs) in the airway extracellular matrix (ECM) remodelling of chronic obstructive pulmonary disease (COPD) rat models. METHODS: The COPD rat models which was established by intratracheal instillation of 200 microg lipopolysaccharide once for every two weeks(twice), and exposed to 5% smoke for 0.5 h/d for 4 weeks. The pathological changes were observed, lung function and blood gas changes were also deter mined. The fibroblasts, lymphocytes of bronchial walls and alveolar macrophages were counted. The hydroxyproline of bronchial lung tissue homogenates were deter mined by biochemistry method. The expression of MMP-9,MMP-2 and tissue inhibitor of metalloproteinase-1(TIMP-1) in bronchi and lung tissue was verfied by immunohistochemical analysis and by reverse transcription-polymerase chain reaction analysis. The gelatinolytic activities of MMPs of lung tissue were performed by gelatin zymographic analysis. RESULTS: The pathological changes of bronchi and lung tissue, the changes of lung function and blood gas analysis were similar to those of the COPD patients. The number of fibroblasts, lymphocytes and alveolar macrophages of model group were significantly increased than those of control group (P < 0.001). The hydroxyproline of model group was significantly in creased than that of control group (P < 0.001). By using image analyzer, immunoreactivity of MMP-9, MMP-2,TIMP-1 were markedly increased in epithelial cells of bronchi, fibroblasts, macrophages, endothelial cells and pneumocytes in model group as compared with those of control group. The protein expressions of MMP-9, MMP-2 and TIMP-1 in model group were significantly increased than those in control group(P < 0.0001 or P < 0.01). The mRNA expression of MMP-2, MMP-9 and TIMP-1 in COPD model group (1.11 +/- 0.06,1.04 +/- 0.26 and 0.85 +/- 0.34,respectively) were significantly increased than those in control group (0.30 +/- 0.17,0.36 +/- 0.09 and 0.23 +/- 0.08,respectively) as well(P < 0.001 or P < "0.01). The relative gelatinolytic activities of 72 000 MMP-2, 92 000 MMP-9 in model group (3 263.5 +/- 665.1 and 1 338.4 +/- 241.2, respectively) were also significantly higher than those in the model group(388.6 +/- 60.8 and 116.1 +/- 49.8,respectively, P < 0.001). CONCLUSION: The findings suggested that there were up regulations of MMP-9,MMP-2 and TIMP-1 of lung tissue in COPD model group which may contribute to the pathogenesis of airflow limitation through the airway remodelling and alveolar structure destruction(emphysema). The ECM degradation and deposition were imbalanced and abnormally activated. The evaluation of MMP-9,MMP-2 which are responsible for the inflammation and destruction process, and the evaluation of TIMP-1 which is responsible for the repair and remodelling process of the airways, may play an important role in the airway ECM remodelling in COPD.


Subject(s)
Matrix Metalloproteinase 2/metabolism , Matrix Metalloproteinase 9/metabolism , Pulmonary Disease, Chronic Obstructive/metabolism , Tissue Inhibitor of Metalloproteinase-1/metabolism , Animals , Blood Gas Analysis , Disease Models, Animal , Electrophoresis, Polyacrylamide Gel/methods , Extracellular Matrix/metabolism , Forced Expiratory Volume , Gene Expression , Lung/metabolism , Lung/pathology , Lung/physiopathology , Male , Matrix Metalloproteinase 2/genetics , Matrix Metalloproteinase 9/genetics , Pulmonary Disease, Chronic Obstructive/genetics , Pulmonary Disease, Chronic Obstructive/pathology , Pulmonary Disease, Chronic Obstructive/physiopathology , RNA, Messenger , Rats , Rats, Wistar , Tissue Inhibitor of Metalloproteinase-1/genetics
13.
Shock ; 17(4): 329-33, 2002 Apr.
Article in English | MEDLINE | ID: mdl-11954836

ABSTRACT

There has been a widespread impression that tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) mediate the toxicity of high doses of lipopolysaccharide (LPS, endotoxin) and are key factors in septic shock. However, the clinical efficacy of treatment with antagonists of TNF-alpha and IL-1beta is still controversial, suggesting that mediators other than TNF-alpha and IL-1beta might contribute causally to endotoxin-induced death. Recent studies implicated high mobility group-1 (HMG-1) protein as a late mediator of endotoxin lethality in mice. However, the role of HMG-1 in mediating multiple organ damage-associating trauma has not been studied. This study was designed to investigate changes in HMG-1 gene expression in vital organs, and its potential role in mediating multiple organ damage following major burns. Wistar rats were subjected to a 35 percent full-thickness thermal injury, and randomly divided into three groups as follows: normal controls (n = 7), thermal injury (n = 24), and recombinant bactericidal/permeability-increasing protein (rBPI21) treatment (n = 12). Tissue samples from liver and lungs were collected to measure tissue endotoxin levels and HMG-1 mRNA expression. In addition, blood samples were obtained for measurement of organ function parameters. Our data demonstrated a significant increase in HMG-1 gene expression in tissues at 24 h postburn, which remained markedly elevated up to 72 h after thermal injury (P< 0.05-0.01). Treatment with rBPI21 could significantly decrease tissue HMG-1 mRNA expression in the liver and lung (P < 0.01). In addition, there were high positive correlations between hepatic HMG-1 mRNA and serum aminoleucine transferase (ALT) and aspartate aminotransferase (AST) levels, and also between pulmonary HMG-1 mRNA and myeloperoxidase activities (P < 0.05-0.01). Taken together, these findings indicate that thermal injury per se can markedly enhance HMG-1 gene expression in various organs. Up-regulation of HMG-1 expression may be involved in the pathogenesis of endogenous endotoxin-mediated multiple organ damage secondary to major burns.


Subject(s)
Burns/genetics , HMGB1 Protein/genetics , Membrane Proteins , RNA, Messenger/genetics , RNA, Messenger/metabolism , Alanine Transaminase/blood , Animals , Anti-Infective Agents/pharmacology , Antimicrobial Cationic Peptides , Aspartate Aminotransferases/blood , Blood Proteins/pharmacology , Burns/complications , Burns/enzymology , Endotoxins/toxicity , Gene Expression , Liver/metabolism , Lung/metabolism , Male , Multiple Organ Failure/etiology , Peroxidase/metabolism , Rats , Rats, Wistar
14.
Zhonghua Shao Shang Za Zhi ; 18(2): 84-7, 2002 Apr.
Article in Chinese | MEDLINE | ID: mdl-12515652

ABSTRACT

OBJECTIVE: To investigate the protective effect and its underlying mechanism of 2,4-diamino-6-hydroxy-pyrimidine (DAHP), an inhibitor of GTP-cyclohydrolase I (GTP-CHI), on postburn Staphylococcus aureus (S. aureus) sepsis in rats. METHODS: Fifty-six Wistar rats were randomly divided into four groups, i.e. normal control, scalding control, postburn sepsis group and DAHP treatment group. Tissue samples from liver, kidneys, lungs and heart were aseptically taken, and in which the GTP-CHI and inducible nitric oxide synthase (iNOS) contents and the mRNA expression of tumor necrosis factor-alpha (TNFalpha) were determined. Furthermore, biopterin (BH(4)) and nitric oxide (NO) levels in these tissue were also measured. RESULTS: After the scalding injury followed by bacterial challenge, the GTP-CHI gene expression and biopterin levels were significantly increased in all tissue sampled, and so were iNOS mRNA expression and NO (P < 0.01), especially in liver and lungs. The expressions of GTP-CHI mRNA and iNOS mRNA and the production of BH(4) and NO in all tissue were evidently inhibited by the pretreatment with DAHP (P < 0.05 approximately 0.01). At the same time, the TNFalpha expression was also obviously decreased. In addition, The mortality at 6 hr in rats of DAHP treatment group was decreased. CONCLUSION: The prognosis of the scalding rats complicated by sepsis caused by G(+) bacteria could be improved by DAHP pretreatment, which might be related to the inhibition of the production of BH(4) and NO by DAHP.


Subject(s)
Burns/complications , Enzyme Inhibitors/pharmacology , Hypoxanthines/pharmacology , Staphylococcal Infections/prevention & control , Staphylococcus aureus/drug effects , Animals , Biopterins/metabolism , Burns/genetics , Burns/metabolism , GTP Cyclohydrolase/antagonists & inhibitors , GTP Cyclohydrolase/genetics , Gene Expression Regulation/drug effects , Heart/drug effects , Kidney/drug effects , Kidney/metabolism , Liver/drug effects , Liver/metabolism , Lung/drug effects , Lung/metabolism , Male , Myocardium/metabolism , Nitric Oxide/metabolism , Nitric Oxide Synthase/genetics , RNA, Messenger/drug effects , RNA, Messenger/genetics , RNA, Messenger/metabolism , Rats , Rats, Wistar , Sepsis/etiology , Sepsis/prevention & control , Staphylococcal Infections/etiology , Staphylococcus aureus/growth & development , Sugar Acids , Time Factors , Tumor Necrosis Factor-alpha/genetics
15.
World J Gastroenterol ; 4(4): 357-359, 1998 Aug.
Article in English | MEDLINE | ID: mdl-11819320

ABSTRACT

AIM:To observe the changes in oxygen free radical (OFR) and the curative effect of traditional Chinese medicine Qing Yi Tang in acute necrotizing pancreatitis (ANP).METHODS:After induction of ANP by injection of sodium taurocholate into pancreatic duct, 16 dogs were randomly divided into control group and Chinese medicine group.Serum amylase, SOD and MDA were determined on postoperative day 1, 2, 4 and 7. The animals were sacrificed on day 7. SOD and MDA in organs were determined, and pathological changes in pancreas were observed.RESULTS: As compared with control group, the serum level of amylase (734U/L vs 2783U/L) and MDA (7.8nmol/ml vs 14.8nmol/ml) in Chinese medicine group were decreased on day 7 (P < 0.05), while SOD increased significantly (281nU/ml vs 55nU/ml, P < 0.01), and similar changes occurred in MDA and SOD in organs, especially in the pancreas; the pathological changes in the pancreas were alleviated as well.CONCLUSION: Qing Yi Tang is effective in clearing OFRs and alleviating pathological changes in ANP.

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