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Objective To explore the effects of polyclonal antibody to lipopolysaccharide binding protein (pLBPab) on IL 10 and IL 18 mRNA expressions of alveolar macrophages in lipopolysaccharide LPS induced acute lung injury in rats. Methods A total of 40 male Wistar rats were randomly divided into 5 groups: normal control (A), LPS treated group (B), group of pLBPab preconditioning at 30 min before injection of LPS (C), group of treatment with LPS and pLBPab (D), and group of pLBPab preconditioning at 30 min after injection of LPS (E). mRNA expressions of IL 10 and IL 18 in the alveolar macrophages in each group were measured by semi quantitative reverse transcription polymerase chain reaction (RT PCR). Results The IL 10 and IL 18 mRNA expressions were highly increased respectively in the alveolar macrophage (AM?) stimulated with single LPS, but they were significantly decreased in the AM? after stimulation with LPS + pLBPab, particularly stimulation with pLBPab 30 min before LPS injection. Conclusion pLBPab can decrease the mRNA expressions such as IL 10 and IL 18 by alveolar macrophages in acute lung injury in rats induced by LPS, and LBP antibody could be used to cure some diseases such as SIRS, acute lung injury, ARDS, and septic syndrome.
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OBJECTIVE: To investigate the classification and incidence of acid-base disturbance (ABD) in the patients with post-traumatic multiple organ dysfunction syndrome (MODS). METHODS: A total of 119 patients with MODS were examined with arterial blood gas analysis and serum electrolytes detection for 675 times in this study. RESULTS: Different types of ABD existed in 647 times out of 675 times (95.9%) of blood-gas analyses. There were 270 times (41.7%) of simple ABD, 271 times (41.9%) of double ABD and 106 times (16.4%) of triple ABD. Among which, 404 times (62.4%) were in respiratory alkalosis (RAL), 332 times (51.3%) in metabolic acidosis (MA), 227 times (35.1%) in metabolic alkalosis (MAL) and 167 times (25.8%) in respiratory acidosis (RA). In this study, 79 cases (66.4%) out of 119 cases with MODS died from these kinds of ABD. CONCLUSIONS: It suggests that in the early stage of MODS, RAL with or without hypoxemia may exist, and later on, MA or even triple ABD may occur. In order to detect and correct the primary disorders as early as possible, it is important to keep the balance of hydrolyte. The treatment of primary diseases is also important. Disorders of acid-base balance were corrected according to pH standard values, anion gap (AG) and the potential [HCO(3)(-)] were also calculated simultaneously. When pH was more than 7.50 or lower than 7.20, it is necessary to give drugs of acidity or alkalinity to the patients with ABD to maintain pH value within a normal range.
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0.05). VIP, however, could inhibit the Con A-induced proliferation of T-cells from control subjects more significantly than that from asthmatics (P0.05). The cAMP level in T-cells, however, increased more significantly in the control group than that in the asthmatic group after the treatment of VIP or NaF (P0.05). CONCLUSION: Inhibition effect of VIP on Con A-induced proliferation of T-cells was less in asthmatics than in control subjects, which may be related to insufficiency of Gs ? coupled VIP receptor on T-lymphocytes in asthmatics.
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AIM: To observe the effects of dexamethasone and interleukin-10 (IL-10) on the release of proinflammatory cytokines, tumor necrosis factor-?(TNF-?), IL-6 and activation of transcriptional factors, nuclear factor-?B(NF-?B), activator protein-1 (AP-1) and cAMP response element binding protein (CREB) in cultrued human peripheral blood mononuclear cells (hPBMC). METHODS: The hPBMC were divided into control group, lipopolysaccharide (LPS) stimulated group, dexamethasone and IL-10 treated group. The contents of TNF-? and IL-6 in supernatant were mensured by ELISA. The activity of NF-?B, AP-1 and CREB of nuclear abstract were analyzed by electrophoretic morbility shift assay (EMSA). RESULTS: The content of TNF-? was significantly increased 1 hour after LPS stimulation, and it was significantly inhibition by dexamethasone and IL-10. The contents of IL-6 and IL-10 were significantly increased after LPS stimulation for 12 hours. The production of IL-6 was still inhibited by dexamethasone and IL-10, but the production of IL-10 was not affected by dexamethasone. The activities of NF-?B, AP-1 and CREB were significantly increased 1 hour after LPS stimulation. Dexamethasone and IL-10 significantly ihibited their activities, but the effects of dexamethasone was stronger than that of IL-10. CONCLUSIONS: LPS induces the release of several pro and anti- inflammatory cytokines and induces the activation of several transcriptional factors in hPBMC. Dexamethasone and IL-10 can inhibite the production of proinflammatory cytokines TNF-?, IL-6 and the activation of NF-?B, AP-1 and CREB. Dexamthasone has more significant inhibitory effect on AP-1 and CREB than IL-10.
الملخص
AIM: To investigate the regulatory effects of lipopolysaccharide binding protein(LBP)on activation of p38 signaling pathway induced by lipopolysaccharide(LPS)in alveolar macrophages. METHODS: The LBP from actue phase rat serum was purified by ammonium sulphate precipitation, Bio-Rex70 resin and the MonoQ column. Rat alveolar macrophages were exposed to LPS (0 01 mg/L or 1 mg/L) the various concentrations of LBP(0 mg/L, 0 01 mg/L, 0 1 mg/L,1 mg/L and 10 mg/L) Western blotting were used to detect phospho-p38 in alveolar macrophages RESULTS: SDS-PAGE analysis indicated that the purified preparation of rat LBP showed homogeneity and the molecular weight was 60 kD.The binding of lipopolysaccharide to mononuclear cells were enhanced by purified rat LBP. Stimulation of rat alveolar macrophages with LPS at concentration of 0.01 mg/L was LBP dependent. LBP at concentrations up to 1 mg/L was able to increase the activation of p38. However , when LBP concentrations were further increased to 10 mg/L, the phosphorylation levers of p38 were lower as compared with that in the presence of 1 mg/L. Stimulation of rat alveolar macrophages with LPS at concentrations of 1 mg/L was LBP-independent. CONCLUSION: The activation of p38 induced by LPS at lower concentration(0.01 mg/L ) was LBP-dependent, meanwhile, LPS at higher concentration (1 mg/L ) was LBP-independent.
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Changes of pulmonary ?_1-and ?-adrenergic receptor (?_1-AR and ?-AR)during endotoxin-induced acute lung injury in rates were oberved to find out the rela-tionship between them and the mechanism of change. Results showed that there was amarked decrease of B_max of both ?_1-AR and ?-AR by 35% and 43% respectively, dur-ing acute lung injury. The down regulation of ?-AR might be one of causes of acutelung injury while that of ?_1-AR seems to be a protective response. Active oxygen playedan important role in endotoxin-induced down regulation of AR in the rat lungs. The increa-sed level of norepinephrine and epinephrine was not the main factor that initiate the downregulation of AR. Intravenous injection of tumor necrosis factor (5?10~6U/kg ) exerts noiafluence on the changes of pulmonary AR in rats.
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AIM: To study the effect of IFN-? inhalation on the anti-infection ability of the lungs in the immunocompromised host. METHODS: The immunological factors in the immunocompromised rats and the immunocompromised rats administrated IFN-? via aerosol were investigated after 1, 3, 7 days when they were injected Candida albicans via tracheal The Canidda albicans count of the left lung was also determined after 7 days when injecting pathogen. RESULTS: The Canidda albicans count of the left lung in IFN-? group was significantly less than that of control group. The phagocyting and bactericidal percentages, Ia antigen expression percentages, the levels of TNF-?, IL-1? and IL-6 in the culture supernatant of the AM, the activity of IFN-? and TNF-? in BALF (except the TNF-? on 7 th day) in IFN-? group were markedly higher than those in control group. The expression of IFN-? and IL-1? pulmonary tissues in IFN-? group was higher than that in control group. The expression of TNF-? in IFN-? group was less than that in control group. The expression of IL-6 was no changes between two groups. The levels of IFN-?, IL-1? and IL-6 in the blood (except IL-1? on 3 rd day), and the killing ability of the lymphocytes in blood had no difference between two groups. CONCLUSION: Administration of IFN-? via aerosol obviously enhanced the anti-infection ability of the lungs in the immunocompromised host, but has no influence on the whole body cellular immunity.
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The changes of erythrocytic membrane band 3 protein and intraery-throcytic and extrar rythrccytic gases and electrolytes were studied in 69 cases of cor pulmonale and 50 normal subjects.It was found that in the patients of cor pulmonale accompanied with type Ⅱ respiratory failure,the relative low level of erythrocytic membrane band 3 protein and the restriction of HCO-3/Cl-exchange were the factors to aggravate CO2 retention and respiratory acidosis,relative intraerythrocytic alkalosis resulted from the relative increase of intra-erythrocytic HCO-3([HCO-3]),and prompt adminstration of oxygen to cor pulmonale patients with hypoxemia could not only improve extraerythrocytic acid-base imbalance but also increase intraerythcocytic P5O2 and the tissue capacity to store oxygen.
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Arterial blood gases,intraerythrocytic pH(pHi),2,3-diphosphoglycerate,standard P50(P50(aid))and in vivo P50(P50iv)were determined in 54 patients with cor pulmonale and in 23 normal subjects.It was found that there was no significant change of pHi but the difference between pHi and extraerythrocytic pH was decreased.P50aid was decreased(P
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Recent evidence suggests Toll like receptor 4 (TLR4) and its accessory protein MD2 mediate endotoxin/lipopolysaccharide (LPS) signaling. LPS binds to LPS binding protein (LBP) in plasma and is delivered to CD14. Next, LPS is transferred to TLR4 MD2 complex. LPS activates several intracellular signaling pathways that include the NF ?B pathway and three mitogen activated peotein kinase (MAPK) pathway: extracellular signal regulated kinase (ERK), c Jun N terminal kinase (JNK) and p38. These signaling pathways in turn activate NF ?B and AP 1 (c fos/c jun), which coordinate the induction of many genes encoding inflammatory mediators. Targeting the signaling molecule in the pathway will develop new remedies for treatment of inflammatory disorder.
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AIM: To explore the mechanism of lipopolysaccharide (LPS) on vascular endothelial cell(VEC) damage. METHODS: By using cytometry techniques, we studied the effects of LPS on apoptosis of human umbilical vein endothelial cells (HUVECs) in vitro . RESULTS: LPS was able to induce apoptosis of HUVECs in a time-dose-dependent fashion.CONCLUSION:Apoptosis might play a role in LPS-induced damage of vascular endothelial cells.
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AIM: To explore effects of LPS,TNF-?,IL-1? on cyclooxygenase-2(COX-2) expression and prostaglandins(PGs) production in human umbilical vein endothelial cells (HUVEC). METHODS: HUVEC were stimulated with lipopolysaccharide(LPS),TNF-?,IL-1? for 24 hours. Using in situ hybridization,reverse-transcription polymerase chain reaction and immunohistochemistry, COX-2 expression in both mRNA and protein level were observed and prostaglandins in culture medium were measured. RESULTS: Resting HUVEC expressed a little COX-2 mRNA. Expression of mRNA and protein of cyclooxygenase-2 increased significantly post-stimulation with LPS,TNF-?,IL-1? and PGs raised markedly at the same time. CONCLUSIONS: Resting HUVEC expressed a little COX-2 mRNA, inflammatory stimuli induced COX-2 superexpression and PGs production. These results suggested that endothelial cells could involve in inflammation by controlling of COX-2.
الملخص
AIM: In order to explore the role of activator protein-1 (AP-1) and cAMP response element binding protein (CREB) in acute lung injury and anti-inflammatory mechanism of glucocorticoid. METHODS: Using the acute lung injury (ALI) model of rats by intravenous lipopolysaccharide (LPS, 5 mg/kg), the activities of AP-1 and CREB by electrophoretic mobility shift assay (EMSA) in lung tissue and effects of dexamethasone on the activities were observed. RESULTS: After injection of LPS, the peak activity of AP-1 was observed at 2 hours and it returned to normal in 12 hours. CREB peak activity was at 1 hours but it did not return to normal in 12 hours. Dexamethasone significantly inhibited the activities of AP-1 and CREB. CONCLUSIONS: AP-1 and CREB may play important roles in ALI in rats. One of the important anti-inflammatory mechanisms of glucocorticoid is its transcription al modification. [
الملخص
Twenty-five hybrid dogs were injected intravenously with oleic acid of the dose 0.3 ml/kg of body weight to prepare a model of respiratory distress syndrome (RDS) . The animals were killed 24 hours after injection. PaO2, PaCO2 and pH of the arterial and mixed venous blood were determined before and immediately, 0.5, 1?2, 4, 6, 22 and 24 hours after injection. The average pulmonary arterial pressure was measured hourly. The chest x-ray films were taken 2,4, 6 and 24 hours after injection. The electrolytes T3, T4, the hematocrit and RBC count, and the serum corticosteroid level were measured before and 24 hours after injection.25 dogs were divided into two groups; the control group consisted of 8 dogs and the therapeutic group consisted of 17 dogs, among which nine were treated with hyosine hydrobromide and 8 with dexamethasone. The histologic specimens of the animals of the control group and hyosine hydrobromide treated group were examined with both light and electron microscopes but the specimens of the animals of dexamethasone treated group were examined with light microscope only.It was found that dexamethasone is effective in the treatment of RDS produced with oleic acid injection while hyosine hydrobromide is of no value.
الملخص
Homogenous fat extract was injected through the femoral vein to induce respiratory distress syndrome in 15 dogs. It was found that the changes of blood gases, chest x-ray films, and lung pathology of the dogs were similar to those of adult respiratory distress syndrome.The pathogenesis was extensive pulmonary fat embolism with complement activation and free radicals formation. Vitamin E was consumed during antiperoxidation. It is believed that this model serves better for the study of respiratory distress syndrome.
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The therapeutic effects of prostaglandin E1 on respiratory distress syndrome induced with homogeneous fat extraction were observed in dogs.It was found that prostaglandin E1 could alleviate hypoxemia,reduce pulmonary capillary permeability,and attenuste pulmonary edema.The mechanism of the therapeutic efficiency of prostaglandin E1 on pulmonary damages is that prostaglandin E1 can inhibit the adherence of polymorphonuclear netttrophils and the genesis of oxygen free radicals,and protect the pneumocyte type Ⅱ.
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The changes of the number of beta-adrenergic receptors, the activity of cAMP-PDE and the level of cAMP were observed in the guinea pigs with allergic asthma with radioligand assay, radioenzyme assay and radioimu-noassay respectively. It was found that there were a 25% decrease of the number of beta-adrenergic receptors on the lung membrane, a 36% decrease of the level of cAMP in the lung tissue, and a 35% increase of cAMP-PDE activity in the asthmatic animals as compared with those of the control. A close positive correlation was observed between the number of beta-adrenergic receptors and cAMP level,but a negative correlation between cAMP-PDE activity and cAMP level in the asthmatic group.
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The levels of CEA, Lys, Fr and ?2-MG of serum and brochoalveolar lavage fluid (BALF) were determined in 99 patients with inflammatory lung diseases and 23 healthy subjects for controls. Their levels in BALF were compared between diseased and normal sides of lungs. The value of diagnosis of individual formulas was suggested. The combined ditermination of multiple biomarkers in serum and BALF can help to raise the positivity and spcificity of diagnosis of lung cancer.
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Twenty-five mongrel dogs were randomized into 3 groups;the animals of group Ⅰ were traumatized with bone marrow extract,those of group Ⅱ were similarily traumatized and then treated with anisodamine,and those of group Ⅲ received saline only and served as control.The specimens of arterial and venous blood were collected and bronchoalveolar lavage fluid (BALF) and lung homogenate were obtained after the animal was killed.The activity of superoxide dismutase (SOD) and the concentration of lipid peroxides (LPO) were determined.Meanwhile,the clinical manifestations,blood gas analysis.chest radiography,and pathological examinations were performed or observed.It was found there were following findings:(1)In group I,SOD activity and LPO level in the lung homogenate and BALF were markedly elevated immediately after injury,which suggests that there had been a rapid production of active oxygen.As a result.various degrees of lung damages were pricipitated.(2)In group Ⅱ,an increase of SOD activity and LPO level in the blood,lung homogenate and BALF:The elevation was more marked than that in group Ⅲ but less marked than that in group Ⅰ.which indicates that there was a relatively of less amount of active oxygen production by the interference anisodamine.(3)In group Ⅱ,no significant changes of SOD activity and LPO livel were found.Our findings suggest that active oxygen is likely to play a very important role in the pathogenesis of acute lung damages in respiratory distress syndrome.
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To explore the effects of lipopolysaccharide (LPS), TNF ? and IL 1? on cyclooxygenase 2 (COX 2) expression and prostaglandins (PGs) in rat polymorphonuclear (PMN), measured COX 2 mRNA was determined by reverse transcription polymerase chain reaction and changes in PGs after stimulation of PMN with LPS, TNF ? and IL 1? for 24 hours were observed. The resting PMN were found to express COX 2 mRNA weakly, while it was significantly up regulated after the stimulation of LPS,TNF ?and IL 1?. The level of PGs was increased markedly at the same time. These results suggested that COX 2 might play an important role during inflammatory process involving PMN.