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Objective To study the effect of plant Coleus forskohlii active elements Isoforskolin(ISOF)and CT-E(analogs mixture of Isoforskolin)on human neutrophill(PMN)in vitro in order to uncover the mechanism of their properties of mitigating acute lung injury(ALI).Method The effects of ISOF and CF-E on PMN aggregation induced by N-formyl-methiony-leucyl-phenylalanine(fMLP)was performed by using a 4-channel platelet aggregometer.Cytometry Was applied to analyze the effect of tested samples on adhension between PMN and endothelial cells(ECV-304)activated by using lipopolysaccharides(LPS).Expression of LPS-induced PMN adhension molecules was determined with flow cytometry.Radioimmunoassay Was applied to detect the level of TNT-α liberated bv PMN and intracellular cyclic adenosine monophosphate(cAMP)level of PMN.Results It was found that ISOF(25,50,100 μmnol/L)and CF-E(1.25,2.5,5 mg/ml)inhibitted PMN aggregation induced by fMLP,PMN adhemion to ECV-304 indeed by LPS,expression of PMN adhesion molecules,and TNF-α level released by PMN.ISOF and CF-E also increased intracellular cAMP level of PMN.Condusions ISOF and CF-E inhibit PMN aggregation,adhension and adhension molecules,and TNF-α released by PMN,while they increase intracellular cAniP level of PMN.It suggests that their specific alleviating the ALI by the mechanism of the modulation of PMN function.
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Objective: To study the effects and significances of lipopolysaccharide preconditioning on the activities of NF-?B and the expression of ICAM-1/LFA-1 in rats graft liver ischemia/reperfusion injury. Methods: Male Sprague-Dawley rats were divided into three groups: sham operation group(Sham group),orthotopic liver transplantation group (OLT group) and LPS preconditioning group (LPS group). Only dissecting hepatoduodenal ligament was perfomed in Sham group. Experiments of OLT were performed by two-cuff method in OLT group and LPS group. The activities of NF-?B and the expression of ICAM-1/LFA-1 in hepatic tissue ,the levels of ALT,AST in inferior caval vein blood were detected at 0,60,180 min after dissecting of hepatoduodenal ligament in Sham group and after portal vein reperfusion in OLT group and LPS group. Results: Compared with those in Sham group at the different time points respectively,the activities of NF-?B and the expression of ICAM-1/LFA-1 were higher in OLT group and LPS group(P0.05) at 0 min after reperfusion,they were evidently higher in OLT group than in LPS group (P
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Objective To investigate the effects of Ginaton on acute lung injury (ALI) induced by lipopolysaccaride (LPS).Methods The acute lung injury rat model was induced by receiving 5 mg/kg LPS injection in tail vein.A total of 63 Wistar rats were divide into 3 groups without caring sex.Saline control group,LPS treated group and Ginaton treated group.The samples of different groups were collected 2 h,6 h,and 10 h after tail vein administration.Serum soluble cell ashesion molecule-1 (sICAM-1) was measured by ELISA,immunohistochemical staining and Western blotting of NF-kB were performed on sections of lung specimens.Results The acute lung injury rat model was successfully induced by receiving LPS injection in tail vein.The sICAM-1 levels increased more in Ginaton treated group than those in Saline control group (P＜0.01),and the increase in LPS treated group were the highest.By immunohistochemical staining,the positive NF-kB cells in Ginaton treated group were much less than those in LPS treated group (P＜0.01),and in Saline control group were least (P＜0.01).The results of the Western-blot method were consistent with immunohistochemical method.Conclusion ALI could be induced by LPS,Ginaton showed a protective effect through probably inhibiting activation of NF-kB on LPS-induced-ALI in this animal model.
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This study was carried out to examine the potency of the three surface components from Porphyromonas gingivalis to stimulate the murine macrophage cell line RAW264.7 to synthesize the pro-inflammatory cytokine tumor necrosis factor alpha(TNF-alpha) and nitric oxide (NO). Lipopolysaccharide(LPS), lipid A-associated proteins(LAP) and saline-extractable surface -associated material(SAM) were isolated from P. gingivalis 381. TNF-alpha release into culture supernatants was determined by two-site ELISA. NO production was assayed by measuring the accumulation of nitrite in culture supernatants. Western blot analysis of iNOS and analysis of reverse transcription(RT)-PCR products were carried out. The surface components extracted from this bacterium were almost equally potent in stimulating release of TNF-alpha and NO by RAW264.7 cells. TNF-alpha that was being measured immunologically was due to activation of TNF-alpha gene transcription. The present study clearly shows that P. gingivalis surface components fully induced iNOS expression in RAW264.7 cells in the absence of other stimuli. The ability of P. gingivalis surface components to promote the production of TNF-alpha and NO may be important in the pathogenesis of inflammatory periodontal disease.
Subject(s)
Tumor Necrosis Factor-alphaABSTRACT
BACKGROUND: Brain inducible nitric oxide synthase (iNOS) might be detectable in several pathologic conditions, and it is thought to play an important role in their pathophysiology. Tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta are believed to be essential factors of iNOS induction of the brain. METHODS: After intrahippocampal stereotaxic injection of lipopoly-saccharide (LPS), the rat brains were removed at 6, 12 and 24 h. The rat brain tissues were examined to clarify the expression patterns of TNF-alpha, IL-1beta and iNOS. RESULTS: The inflammatory cells which were stained with anti-TNF-alpha antibody, appeared in 6 h and increased for 24 h after LPS injection. The iNOS positive cells appeared after 12 h of LPS injection. A semiquantitative analysis of reverse transcription-polymerase chain reaction (RT-PCR) revealed that the TNF-alpha and IL-1beta mRNA arose at 1 h, peaked at 6 h and then declined until 48 h after LPS injection. The iNOS mRNA arose after 6 h, peaked at 12 h, and declined until 48 h after LPS injection. CONCLUSIONS: We conclude that the induction of inflammatory events by intrahippocampal injection of LPS activates TNF-alpha and IL-1beta secretion, and this is followed by an induction of iNOS expression. TNF-alpha and IL-1beta seem to be related with iNOS expression in brain inflammation.
Subject(s)
Animals , Rats , Brain , Encephalitis , Hippocampus , Interleukin-1beta , Interleukins , Nitric Oxide Synthase Type II , RNA, Messenger , Tumor Necrosis Factor-alphaABSTRACT
Objective To investigate whether gingko biloba extract (GBE) have protective effect on acute lung injury(ALI)induced by Lipopolysaccharide ( LPS) in aging rats and renal function damage induced by ALI. Methods Male aging Wistar rats(36) were used in the study. LPS group (LPS, iv route, 5mg/kg body weight) and GBE+LPS group (GBE was given starting from 7 days before experiment, once a day oral administration). The blood, lung and kidney samples were collected at 2 or 6 hours after LPS or saline administration. Results Compared with the aging control, ALI was obviously observed in LPS group, blood creatinine and urea nitrogen were increased significantly from (68.7? 6.9) mol/L and (5.9?0.6) mmol/L to (94.7?10.3) mol/L and (11.4?1.9) mmol/L, respectively (P