AIM:To investigate LPS(lipopolysaccharide)stimulated cytokine secretion from normal rat kupffer cells in vitro. METHODS: Kupffer cells were isolated from wistar rats liver and cultured. Tumor necrosis factor - α (TNF- α) and endothelin- 1 (ET- 1 ) secreted by LPS stimulated kupffer cells were detected. RESULTS: LPS had an stimulative effect on kupffer cell activity. LPS in definite concentrations promoted kupffer cell secretion. CONCLUSION: LPS promotes kupffer cell secretion, which may be associated with liver injury induced by LPS.

AIM:To investigate LPS(lipopolysaccharide)stimulated cytokine secretion from normal rat kupffer cells in vitro. METHODS: Kupffer cells were isolated from wistar rats liver and cultured.Tumor necrosis factor -? (TNF-?) and endothelin-1(ET-1) secreted by LPS stimulated kupffer cells were detected. RESULTS: LPS had an stimulative effect on kupffer cell activity. LPS in definite concentrations promoted kupffer cell secretion.CONCLUSION: LPS promotes kupffer cell secretion, which may be associated with liver injury induced by LPS.

AIM:The objective of the study was to explore whether intestinal endotoxemia participate in the development of Alzheimer disease.METHODS:Adult Wistar rats were subjected to 90 days intraperitoneal injection with D-galactose and aluminum trichloride(AlCl3) to establish the model of Alzheimer disease.After the administration,the study and memory ability in the rats were observed by Morris water maze.The level of lipopolysaccharide(LPS) in the sera of Alzheimer disease's rats was determined by tachypleus amebocyte lysate method.The level of tumor necrosis factor-?(TNF-?) and interleukin-1(IL-1) in the sera were determined by radioimmunoassay.The expressions of amyloid ?-protein precursor(APP),presenilin 1(PS1) and ?-site APP-cleaving enzyme(BACE) in hippocampus were detected by RT-PCR.RESULTS:Compared with the normal control,the level of LPS in the sera and the expressions of APP,PSI,BACE mRNA in the hippocampus were markedly increased(P

Objective To investigate the effect of hepatic microcirculatory disturbance induced by intestinal endotoxemia in liver injury. Methods The model of rats with intestinal endotoxemia induced by thioacetamide(TAA) was established. 25 Male Wistar rats were divided into 4 groups as normal control group (N), Heparin control group(H), TAA treated group(T), and TAA + heparin treated group(T +H). The changes of plasma biochemistry and hepatic histopathology were measured. Results The plasma endotoxin level, alanine transaminase (ALT) activity and malondialdehyde (MDA) content in TAA treated rats were markedly higher than that in the control ones (P < 0.05), while plasma endotoxin level was lower ( P > 0.05) and ALT as well as MDA were decreased significantly ( P < 0.05) in TAA + heparin group than in TAA group. Conclusion Intestinal endotoxemia could induce disturbance of hepatic microcirculation, which results in ischemia and hypoxia of liver cell. Heparin could not only correct disturbance of hepatic microcirculation induced by intestinal endotoxemia, but also reduce liver injury induced by ischemia and hypoxia.

AIM:To study the effect of intestinal endotoxemia(IETM) on hepatic energy metabolism in acute liver failure. METHODS:Intoxication by thioacetamide (TAA) was used to establish rat model of acute liver injury.Ketone body(acetoacetate and ?-hydroxybutyrate) in arterial blood and ATP content of hepatocellular mitochondria were determined by using enzymatic fluorimetric micromethod.Colectomy was adopted in observing the changes in plasma endotoxin content and serum alanine aminotransferase (ALT) activity. RESULTS:In the TAA group,plasma endotoxin content and serum ALT activity were all significantly higher than those in the control group(P

Objective To investigate the relationship between dendritic cell (DC)and intestinal endotoxemia in patients with chronic hepatitis B (CHB).Methods Peripheral blood were collected from CHB patients (n = 80)and healthy controls (n = 21 ).Plasma endotoxin (ET)levels,liver function (alanine transaminase,total bilirubin)were detected.According to plasma ET concentration,all CHB patients were divided into two groups:ET positive and ET negative.The peripheral blood mononuclear cells (PBMCs)were isolated and then cultured with recombinant human granulocyte-macrophage colony-stimulating factor ( rhGM-CSF),recombinant human interleukin-4 ( rhIL-4 ),FMS-related tyrosine kinase 3 ligand (Flt3L)and tumor necrosis factor-alpha (TNF-α)to derive DC.The phenotypic patterns were characterized by flow cytometry.The proliferation of T lymphocytes was evaluated with mixed leukocytes reaction (MLR)and the levels of IL-12 and interferon-γ (IFN-γ)produced by DC were analyzed with enzyme-linked immunosorbent assay (ELISA).Comparisons among the two groups and healthy control group were done by single factor analysis of variance.Results Compared to healthy controls,the expressions of CD83,CD80,CD86,human leucocyte antigen (HLA)-DR and the proliferation of allogeneic T lymphocytes by DC were all significantly reduced in CHB patient groups.The expressions of CD83,CD80,CD86,HLA-DR and the activation of proliferation in ET positive subjects were lower than those in ET negative subjects [CD83 (8.25±3.63)% vs(11.39±4.35)% ,CD80 (10.63±4.52)% vs (13.56±5.13)%,CD86 (36.61±16.16)% vs (45.90±15.35)%,HLA-DR (61.65±14.33)% vs (70.35±18.89)%,the activation of proliferation0.812±0.311 vs 1.153±0.324; F=5.123,4.213,3.714,3.323 and 3.125,respectively; all P＜0.05].After cultured for 9 days,the secretions of IL-12 and IFN-γ by DC were significantly lower in CHB patients than in healthy controls [IL-12 (16.99± 6.74)pg/mL vs (44.51±14.56)pg/mL,IFN-γ (10.52±4.19)pg/mL vs (17.94±5.86)pg/mL].The level of IL-12 in the ET positive group was significantly lower than that ET negative group [( 13.14 ±5.71)pg/mL vs (20.98 ± 9.03)pg/mL; F= 3.225,P = 0.016].The level of IFN-γ was not different between two groups [(9.46 ± 3.24)pg/mL vs (11.54 ± 5.20)pg/mL; F = 2.003,P =0.076].Conclusion The intestinal endotoxemia may play a role in DC dysfunction in CHB patients.

Objective To evaluate the effect of anti-histamine treatment on intestinal endotoxemia and liver inflammation in experimental chronic hepatitis rats.Methods Thirty Wistar rats (15 males and 15 females) were randomly divided into control group (n =8),chronic hepatitis group (n =12) and hepatitis + anti-histamine group (n =10).Chronic hepatitis was induced by subcutaneous injection with 40％ of CCl4,and feeding with low protein,low choline,high cholesterol and high alcohol diet.Antihistamine treatment was given 1 week after the modeling by intragastric administration of ketotifen (1.25 mg/kg).All rats were sacrificed 4 weeks later.Plasma endotoxin,alanine aminotransferase (ALT),total bilirubin (TBil),tryptase,histamine,interferon-γ (IFNγ),iuterleukin (IL)-12,IL-10 and IL-4levels were detected,and the changes in liver histology,the morphology and ultrastructure of mast cells were observed.SPSS 13.0 software package was used for statistical analysis.ANOVA was used for the comparison of measurement data,and SNK method was used for pairwise comparison.Results Plasma endotoxin,ALT,TBil,tryptase,plasma and liver tissue histamine concentrations were (81 ± 19) pg/mL,(186 ± 140) U/L,(10.2±6.2) μmol/L,(0.75 ±0.21) mg/mL,(145 ±52) ng/mL,and (107 ±43) ng/100 mg in chronic hepatitis group,while the above parameters were significantly lower in anti-histamine group except TBil (P ＜ 0.05).Under light microscope,fatty degeneration and fibrosis were formed in liver of chronic hepatitis rats,the hepatic injury was attenuated in anti-histamine group.Toluidine blue stain showed that there was many degranulating and degranulated mast cells filled with purple granula around liver blood vessels and in fiber-interval in chronic hepatitis group,and there were few purple granula in anti-histamine group.The number of mast cells in anti-histamine group was (6.5 ± 1.5)/HP,which was significantly lower than chronic hepatitis group [(10.9 ± 1.6)/HP,P =0.000],but was still higher than that in the control group [(2.2 ± 0.9)/HP,P =0.000].Under electron microscope,the phenomenon of degranulation was severe in chronic hepatitis group and moderate in the anti-histamine group.Compared with the chronic hepatitis group,IL-4 and IL-10 in anti-histamine group were significantly decreased (P ＜0.05),IL-12 was increased (P ＜0.05),but the level of IFN-γ had no significant change (P ＞ 0.05).Conclusion Anti-histamine therapy can significantly improve liver inflammation and alleviate intestinal endotoxemia.

AIM:To investigate the effect of lipopolysaccharides(LPS)preconditioning on CCl4-induced liver injury and the change of LPS signal transduction.METHODS:The male Wistar rats were divided randomly into liver-injury group,which were injected with CCl4 5 mL/kg first,three days later were injected 0.3 mL 40% CCl4 and 60% olive oil. Animals in LPS preconditioning group were injected with LPS 0.5 mg/kg before the day CCl4 was given. Rats received high fat diet were as liver injury group,and normal control group received normal diet. The lymphocytes infiltrated in the liver tissue were counted. The endotoxin and ALT level in rat plasma,TNF-? content and expressions of TLR4,p38,p-p38,I??,NF-?? in the rat livers were also determined.RESULTS:The lymphocytes in liver slice and ALT level of the plasma in LPS preconditioning group were lower significantly than those in the liver injury group,and the expressions of TLR4,p-p38,NF-?? in the liver were the same. In contrast,the expression of I?? was higher.CONCLUSION:LPS preconditioning relieves obviously CCl4-induced chronic liver injury. The mechanism may be associated with change of signal transduction of LPS,which results in decrease of pre-inflammatory cytokines.

AIM: To investigate the effect of LPS on phagocytosis of Kupffer cells in vitro. METHODS: Isolated Kupffer cells were treated with LPS in vitro . The phagocytosis, microfilament, microtubules and apoptosis of Kupffer cells were determined by the beads phagocytosis test, fluorescence staining, fluorometry and flow cytometric analysis. RESULTS: LPS enhances the phagocytosis, actin content, microtubules fluorescence density of Kupffer cells in vitro , while at a large dose or for a long time, it lessened the phagocytosis increasing or phagocytosis, inhibites the microfilament and microtubules expression, and induced apoptosis. CONCLUSION: LPS enhances the phagocytosis of Kupffer cells in vitro , but in large amount, it inhibites the phagocytosis of Kupffer cells, which is probably related to LPS -induced microfilament, microtubules expression changes and apoptosis in Kupffer cells.

Objective To investigate the effects of Shuangligan and Glycine on Thl/Th2 balancing on severe acute pancreatitis ( SAP) in rats. Methods Thirty-two Wistar rats weighing (260 ± 20) g were randomly divided into sham operation (SO) group, SAP group, SAP + Slg (with the treatment by Shuangligan) group and SAP + Gly (with the treatment by Glycine ) group. Each group included 8 rats, which accepted different treatment according to the experimental design. Changes of plasma level of endotoxin ( ET) and serum amylase (AMY) and the effects of Shuangligan and Glycine on Thl/Th2 ratio at the 24th hour after operation were observed respectively. Results The plasma endotoxin (ET) level ( (0. 67 ±0. 11) EU/ml),proinflammatory cytokine (INF-γ:(8.43 ± 0.86) ng/L, IL-12: (8.26 ± 1.97) ng/L) and Thl/Th2 ratio (0.36 ± 0.07) in SAP group were significantly higher than those in SO group( ET: (0. 44 ±0.07) EU/ml, INF-γ: (3. 80 ±0. 55) ng/L, IL-12: (3. 34 ± 1. 34)ng/L,Thl/Th2 ratio (0. 24 ±0. 05) ) (P <0. 05). Compared with SAP group, SAP + Slg and SAP + Gly group had remarkably decreased plasma ET level ( (0. 57 ± 0. 08,0. 52 ± 0. 04) EU/ml) (P < 0. 05) and the Thl/Th2 ratio reached equilibrium ( SAP + Slg group; (0. 29 ± 0. 04 ), SAP + Gly group: (0. 25 ± 0. 06 )) . Conclusions In the earlier stage of SAP, the rising plasma ET level may cause the overreaction of the cell mediated immune response, which leads to the aggravated damages in tissue cells. Our data indicates that Shuangligan and Glycine can restrain the formation of intestinal endotoxemia and alleviate or prevent the tissue injuries.