Curcumin attenuates lipopolysaccharide/D-galactosamine-induced acute rat liver injury via activating autophagy of hepatocytes

AIMTo investigate the effect of curcumin (CUR) on autophagy of hepatovyte in rats with lipopolysaccharide (LPS)/D-galactosamine (D-GalN)-induced acute liver injury (AHI). METHODSThe healthy male Sprague-Dawley (SD) rats were randomized into the control group,AHI group,CUR group, 3-methy-ladenine (3-MA) group and 3-MA+CUR group, with 6 rats in each group. AHI was induced with an intraperitoneal injection of LPS and D-GalN. Liver function was tested 12 h after LPS/D-GalN treatment. Pathological changes of liver tissues were analyzed by HE staining.The amount of autophagic bodies were observed by transmission electron microscopy. The protein levels of autophagy related-proteins LC3 and beclin 1 in livers were detected by Western blot. ELISA were used to examine the serum levels of tumor necrosis factor-α (TNF-α). RESULTSCompared with control group, the serum level of alanine aminotransferase (ALT) and asparated aminotransferase (AST) were significantly increased, hepatic pathological damage were aggravated and serum TNF-α level was significantly increased in AHI group, while the autophagic bodies and the protein levels of LC3 and beclin 1 were increased (P<0.01). Compared with AHI group, the serum level of ALT and AST were significantly decreased, hepatic pathological damage were attenuated and serum TNF-α level was significantly reduced (P?<0.05), while the autophagic bodies and the protein levels of LC3 and beclin 1 were significantly increased in CUR group (P<0.01). Compared with CUR group, the serum level of ALT and AST were significantly increased, hepatic pathological damage were aggravated and serum level of TNF-α was significantly increased in 3-MA group and 3-MA+CUR group, while the autophagic bodies and the protein levels of LC3 and beclin 1 were decreased (P<0.01). CONCLUSION Curcumin protects rats against LPS/D-GalN-induced liver injury, partially due to activation of hepatocyte autophagy in livers.     

Mechanisms for a decrease in mortality of LPS-challenged mice by rhynchophylline

AIM: To observe effect of rhynchophylline （Rhy） on mortality and organ injury in endotoxemic mice and further investigate the mechanisms of its actions. METHODS: Male mice were randomly assigned into control, LPS, Rhy +LPS and Rhy group, and injected subcutaneously with normal saline (0.05 mL/10 g), or rhynchophylline once a day for 3 d, 1 h after subcutaneously treatment on day 3, LPS (20 mg/kg) or normal saline was injected intraperitoneally. Survival rate was recorded every 12 h for 6 d. In another experiment, 12 h after LPS injection, the left lung and intestine tissue sections were prepared for histological analysis and the right lung were used to determine the ratio of wet to dry lung tissue weight (W/D),the serum was collected to detect the concentrations of alanine aminotransferase（ALT）, aspartate aminotransferase (AST ), bloodureanitrogen (BUN) and creatinine (Cr). In addition, the concentrations of tumor necrosis factor-α (TNF-α), interleukin-1β(IL-1β) and interleukin-10 (IL-10) in serum at 2 h after LPS challenge were detected by enzyme-linked immunosorbent assay. The concentration of NO in serum at 8 h was detected by enzymic method. The effect of Rhy on survival rate of mice subjected to cecal ligation and puncture (CLP) was also observed. RESULTS: Mortality of mice challenged with LPS alone was higher significantly than that in control at 24 h after LPS challenge, pretreated with Rhy at a dose of 8 or 16 mg/kg increased markedly the survival rate of LPS-challenged mice. However, Rhy at a dose of 8 mg/kg significantly increased mortality of mice subjected to CLP. In the histological analysis, severe inflammation was observed both in the lung and intestine tissues in the LPS group. LPS elevated lung W/D, the levels of ALT, AST, BUN, Cr, TNF-α, IL-1β, IL-10 and NO in serum. Pretreatment with Rhy had no obvious improvement in the lung and intestine tissue injury, and no significant depression in the lung W/D and the serum levels of ALT, AST, BUN, Cr, IL-1β, IL-10 and NO, but decreased the level of TNF-α in serum significantly in LPS -treated mice. CONCLUSION: Pretreatment with Rhy reduces the mortality in endotoxemic mice, but not decrease the mortality of mice challenged with CLP, at least in part, through inhibiting the synthesis and secretion of TNF-α.     

Mechanisms underlying the protection of berberine against liver injury induced by lipopolysaccharide in mice

AIM: To investigate the protective effects of berberine against liver injury induced by lipopolysaccharide in mice and the mechanisms underlying its protective effect. METHODS: The male mice were divided randomly into control, berberine group, LPS group and berberine treatment group. Mice were administered intragastrically with distilled water (0.01 mL/g) or 5 g/L neutral sulfate berberine (0.01 mL/g) once a day for 5 days and injected intraperitoneally with normal saline or LPS （0.02 mL/g，28 mg/kg）at 1 h after gavage on day 5. Blood was collected for determining alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, the content of tumor necrosis factors-α (TNF-α) at 10 h and 2 h after LPS or normal saline injection, respectively. Furthermore, the liver tissue was processed, and histological changes and ultrastructure in liver were observed with light and electron microscopy, malondialdehyde (MDA) content and superoxide dismutase (SOD) activity in liver were also detected. RESULTS: Both ALT and AST activities in serum in LPS group were higher than those in control and berberine treatment group. LPS increased the serum TNF-ɑ content at 2 h after injection, which was reversed by berberine pretreatment. The histological examination showed that LPS caused severe hepatic cell edema, degeneration, apoptosis and even necrosis, and ultrastructure observation demonstrated that LPS induced mitochondrial swelling, condensation and margination of chromatin, irregular nuclear envelope in hepatocytes. The above pathological changes produced by LPS were attenuated by berberine pretreatment. Moreover, MDA contents in liver tissue were higher in LPS group than control and berberine treatment group, but there were no significant difference in SOD activity between berberine treatment and LPS group. CONCLUSION: Berberine has a protective effect on LPS-induced liver injury in mice, the mechanisms may be related to its decreasing the production of TNF-α, inhibiting lipid peroxidation and protecting mitochondria.     

不同浓度氮素对翠菊幼苗质量的影响

以硝酸钙为氮源,采用单因素随机区组试验设计,设5个氮素浓度,研究氮肥对翠菊幼苗生长的影响。结果表明:硝酸钙用量为125 mg/kg基质时,翠菊幼苗的干重与其它处理间呈极显著差异;硝酸钙用量在1001、25 mg/kg基质水平时,翠菊幼苗的茎粗达到最大值;硝酸钙用量水平为125 mg/kg基质的处理壮苗指数达到最大;氮素用量对翠菊幼苗植株高度影响不明显。     

Effects of silymarin on LPS-induced acute lung injury in rats

AIM:To investigate the effects of silymarin on lipopolysaccharide (LPS)-induced acute lung injury in rats and its possible molecular mechanisms. METHODS:Fifty-eight male SD rats, weighting 230-250 g, were divided into four groups randomly: normal control (n=12); acute lung injury group (n=15), receiving intravenous LPS (O55∶〖KG-*2〗B5, 5 mg/kg); silymarin alone group (50 mg/kg, n=15); intervention group (n=16, receiving silymarin 50 mg/kg and LPS 5 mg/kg). The specimens were collected 6 hours later. The following changes, including blood gas analysis, the lung wet/dry weight ratio, the pulmonary vascular permeability, histological manifestations, lung tissue myeloperoxidase activity, the levels of TNF-α, IL-1β, MCP-1 and SOD, GSH-Px as well as malonaldehyde and conjugated diene in plasma and lung tissue, were observed. RESULTS:Compared with control group, the lungs of the rats in LPS treatment group showed significant hyperemia and spotted hemorrhage. The inflammatory granulocyte infiltrating, diffused alveolar septum thickening and spotted hemorrhage were observed in pathological examinations. The lung wet/dry weight ratio and Evans blue content (per gram) increased significantly after LPS treatment. The myeloperoxidase activity in plasma and lung tissue, the levels of TNF-α, IL-1β, MCP-1 and SOD, GSH-Px as well as malonaldehyde and conjugated diene were increased significantly in LPS treatment group. However, in intervention groups, all the above-mentioned measurements were reversed significantly by silymarin treatment compared with LPS treatment group. CONCLUSION:Silymarin may decrease inflammatory reaction and oxidative stress, and further decrease lung damage induced by LPS in rats, all indicating protection of silymarin against acute lung injury.     

Astragaloside Ⅳ inhibits JAK2/STAT3 signaling pathway and alleviates severe acute pancreatitis-associated acute liver injury in rats

AIM: To investigate the effect of astragaloside Ⅳ on severe acute pancreatitis (SAP)-associated acute liver injury in the rats and to explore the underlying mechanisms. METHODS: Male Sprague-Dawley (SD) rats (n=96) were randomly divided into sham-operated group, SAP model group, astragaloside Ⅳ treatment group and AG490 treatment group. SAP model was induced by retrograde injection of 5% sodium taurocholate (1 mL/kg) into the biliopancreatic duct. The rats in astragaloside Ⅳ treatment group were intraperitoneally injected with 20 mg/kg astragaloside Ⅳ, while the rats in AG490 treatment group were injected with 8.0 mg/kg AG490 2 h before sodium taurocholate injection. The rats in sham-operated group and model group received the same volume of saline. The rats were sacrificed at 12 h, 18 h and 24 h after the treatment. The levels of ascites, serum amylase, ALT and AST were detected after the blood samples were collected by the puncture through inferior vena cava. The serum levels of TNF-α, IL-6 and IL-1β were also examined by ELISA. Furthermore, HE staining was used to observe the liver pathological changes, and the protein levels of p-JAK2 and p-STAT3 in the liver were evaluated by Western blot. RESULTS: Compared with sham-operated group, the levels of ascites, serum amylase, ALT, AST, IL-6, TNF-α and IL-1β in the rats in model group were significantly increased, while they were decreased in the rats in astragaloside Ⅳ treatment group and AG490 treatment group compared with the rats in model group. Meanwhile, the phosphorylation levels of JAK2 and STAT3 was significantly increased in model group compared with sham-operated group. The rats in astragaloside Ⅳ treatment group and AG490 treatment group both had a better improvement in the liver injury and lower phosphorylation levels of JAK2 and STAT3.CONCLUSION: Astragaloside Ⅳ exerts a protective effect on pancreatitis-associated acute liver injury in the rats possibly via inhibiting JAK2/STAT3 signaling pathway.     

LPS preconditioning relieves chronic liver injury induced by carbon tetrachloride

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-α content and expressions of TLR4, p38, p-p38, IκΒ, NF-κΒ 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-κΒ in the liver were the same.In contrast, the expression of IκΒ 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.     

Protective effect of low dose of hydrocortisone on the liver in LPS attack rats

AIM:To study the effect of different dosage of hydrocortisone on the liver in lipopolysaccharides(LPS) attack rats.METHODS:The model of LPS attack rats was established,and different doses of hydrocortisone were given to the rats. ALT and AST levels in rat plasma were tested,and the histology of rat liver was observed by microscope. RESULTS:ALT and AST levels were high in LPS group and had significant difference compared with the normal control group. ALT level in low dose(LD) group had no significant difference compared with the normal control group. The pathological change in the liver was obviously congested in high dose(HD) group and LPS group,many inflammatory cells were infiltrated. The change of liver in LD group was slight. CONCLUSION:Low dose hydrocortisone may have the protectiive effect on liver in LPS attack rats. High dose and middle dose of hydrocortisone have no effects.     

Effects of ouercetin on lipopolysaccharide induced hepatocyte injury in vitro

AIM: To study the effects of quercetin on lipopolysaccharide (LPS) induced hepatocyte injury and the expression of TNF-α in vitro. METHODS: Hepatocytes were isolated from male Sprague-Dawley rats by collagenase perfusion. LPS at concentration of 40 mg/L was used to induce injury to the cultured cells, and 0.5-10 μmol/L quercetin was added at the same time. After 24 h of incubation, the cell apoptosis rates were detected by MTT and PI-AnnexinV. LDH and TNF-α were measured by kits. RESULTS: 40 mg/L LPS caused a 27% growth inhibition. The apoptosis rate was 30.2%. LDH leakage was 20 folds higher than normal. TNF-α expression significantly increased. Treated with quercetin at doses of 0.5-10 μmol/L, the apoptosis rate, LDH leakage and TNF-α expression in hepatocytes were attenuated in a dose dependent manner. CONCLUSION: 0.5-10 μmol/L of quercetin protects hepatocytes from injury induced by LPS, which is associated with suppression of the inflammatory cytokine TNF-α.     

Effects of propofol on expression of AIF and cell apoptosis in brain tissues of the rats with LPS-induced brain injury

AIM: To investigate the effects of propofol on the expression of apoptosis-inducing factor (AIF) and cell apoptosis in brain tissues of rats with lipopolysaccharide(LPS)-induced brain injury. METHODS: Seventy-two male and female SD rats weighing 220~250 g were randomly divided into 3 groups (n=24 each). Cerebral edema was induced by injection of LPS at 1 mg/kg via left internal carotid artery in LPS group and LPS+propofol group. In control group, equal volume of normal saline was administered instead of LPS. The rats in LPS+propofol group received intraperitoneal injection of propofol at 100 mg/kg immediately after LPS administration. Six rats in each group were decapitated 6 h, 12 h, 24 h or 48 h after operation and the frontal lobe cortex were immediately removed for determination of the water content. The apoptotic neurons were detected by Annexin V-PI staining. The protein levels of AIF, NF-κB and caspase-3 were measured by immunohistochemistry. The protein expression of AIF was detected by Western blotting analysis. RESULTS: Compared with control group, the brain water content, the number of neuronal apoptosis and the protein expression levels of AIF, NF-κB and caspase-3 were significantly increased in LPS group and LPS+propofol group. Compared with LPS group, the results mentioned above were markedly reduced in LPS+propofol group. CONCLUSION: Propofol attenuates LPS-induced brain injury by decreasing AIF protein expression and inhibiting apoptosis.     

Effects of neutrophil extracellular traps on acute lung injury in neonatal rats

AIMTo investigate the role of neutrophil extracellular traps (NETs) in neonatal rats with acute lung injury (ALI). METHODSThirty 7-day-old SD rats were randomly divided into normal saline control group, ALI group and ALI+deoxyribonuclease (Dnase) group (each n=10). The rats in ALI group were intraperitoneally injected with lipopolysaccharides (LPS) at 20 mg/kg, and the rats in ALI+Dnase group were intraperitoneally injected with Dnase at 5 mg/kg after LPS injection. After 6 h, the rats were anesthetized with chloral hydrate, bronchial alveolar lavage fluid (BALF) was collected, and the content of cell-free DNA (cf-DNA) in BALF was detected by fluorescence microarray. The right lung tissues were fixed in 4% paraformaldehyde, and the morphological structure of the lung tissues were observed by HE staining. The content of interleukin-6 (IL-6) and tumor necrosis factor-α （TNF-α） in the left lung homogenate was measured by ELISA. Immunofluorescence and Western blot were used to detect the production of citrullinated histone H3 (CitH3) and myeloperoxidase (MPO) in the rat lung tissues. RESULTSCompared with control group, the levels of cf-DNA, CitH3, MPO, IL-6 and TNF-α in lung tissues of neonatal rats in ALI group and ALI+Dnase group were all increased (P<0.05), and severe inflammatory infiltration in the lung tissues was observed. Compared with ALI group, the levels of cf-DNA, CitH3, MPO, IL-6 and TNF-α in ALI+Dnase group were decreased (P<0.05), and the inflammatory infiltration was attenuated. CONCLUSION In neonatal rats with ALI, the level of NETs is an important indicator of lung tissue injury, and NETs may be a new target for the treatment of neonatal ALI.     

Effects of GW1929 on macrophage TLR expression and inflammation induced by ox-LDL

AIM: To investigate the effects of ox-LDL on TLR2 and TLR4 expression and production of TNF-α, IL-10, IL-12, NO and MDA in macrophages and to observe intervention effect of GW1929 in above procedure. METHODS: The mouse peritoneal macrophages were pretreated with ox-LDL (50 mg/L, 100 mg/L) and GW1929 (20 μmol/L) respectively for 24 h. The concentrations of MDA, NO-2/NO-3, TNF-α, IL-10 and IL-12 in the culture fluid were detected. Flow cytometry was used to observe TLR2 and TLR4 expressions after the mouse peritoneal macrophages were pretreated with ox-LDL (50 mg/L) and GW1929 (20 μmol/L) respectively for 6 h, 12 h, and 24 h. RESULTS: The concentrations of MDA, NO-2/NO-3, TNF-α and IL-10 in ox-LDL (50 mg/L, 100 mg/L) group were higher than those in control and GW1929 group obviously, but the concentrations of above index in ox-LDL (50 mg/L, 100 mg/L)+GW1929 group were lower than those in ox-LDL (50 mg/L, 100 mg/L) group apparently. No IL-12 in every group was detected. Expressions of TLR-2 in ox-LDL+GW1929 (6 h, 12 h, 24 h) group were lower than those in ox-LDL (6 h, 12 h, 24 h) group respectively. TLR-4 expressions in ox-LDL+GW1929 (12 h) were lower than those in ox-LDL (12 h) apparently. CONCLUSION: ox-LDL up-regulates TLR2 and TLR4 expressions and promotes the production of ROX, NO, TNF-α and IL-10 in macrophages. GW1929 is capable of inhibiting the above ox-LDL effects.     

Effects of thalidomide on the expression of NF-κB and TNF-α in experimental hepatic fibrotic rats

AIM: To study the effects of thalidomide on the expressions of nuclear factor κB (NF-κB) and tumor necrosis factor-α (TNF-α) in rat liver fibrosis.METHODS: The fibrosis of rat liver was induced by intraperitoneal injection of carbon tetrachloride thrice weekly.Meanwhile thalidomide (10 mg·kg^-1·d^-1 or 100 mg·kg^-1·d^-1) was given daily by the intragastric route for 8 weeks.Serum alanine aminotransferase (ALT),aspartate aminotransferase (AST),prealbumin (PA),hyaluronic acid (HA) and laminin (LN),and hydroxyproline (HYP) contents in the liver,NF-κB p65 and α-smooth muscle actin (α-SMA) protein in the liver,IκBα and TNF-α protein in cytoplasm and NF-κB p65 protein in nucleus and TNF-α mRNA levels in the liver were studied.RESULTS: Compared with the model group,the Knodell score,serum ALT,AST,HA,LN levels and HYP contents in liver,NF-κB p65 protein in nucleus and α-SMA protein in the liver,and TNF-α mRNA and protein in the liver of rats given high dose of thalidomide were decreased significantly (P<0.01).Meanwhile PA level and IκBα protein in cytoplasm were elevated significantly (P<0.01).CONCLUSION: Thalidomide exerts its effect on the down-regulation of NF-κB-induced TNF-α via inhibiting dissociation and degradation of IκB and prevents liver fibrosis in rats.     

Role of IRAK-4 activity in inhibitory effects of lipopolysaccharide pretreatment on hepatic ischemia/reperfusion injury

AIM:To observe the changes of interleukin-1 receptor associated kinase-4 (IRAK-4) in ischemia/reperfusion (I/R) liver pretreated with lipopolysaccharide (LPS) and to explore the protective mechanisms of LPS pretreatment against hepatic I/R injury. METHODS:Male Sprague-Dawley rats, weighing 240-280 g, were divided into three groups:control, ischemia/reperfusion group (I/R group) and LPS-pretreated group (LPS group). On the first day, LPS group received 0.1 mg/kg LPS via the tail vein, followed by 0.5 mg/kg on the 2nd, 3rd, 4th and 5th day. I/R group received the equivalent volumes (0.5 mL) of sterile PBS. Experiments of I/R injury was induced by temporary ischemia of the left lateral liver lobe for 90 min followed by 3 h reperfusion on 2 days after the last LPS treatment. At 0 min, 60 min and 180 min after reperfusion, the expression of IRAK-4 gene and protein level were determined by RT-PCR and Western blotting. The activity of NF-κB and the serum TNF-α level were also detected by ELISA. RESULTS:Although the level of IRAK-4 gene and protein were higher in the LPS group than that in I/R group and control group (P<0.01), no difference of the activities of NF-κB and the TNF-α level was observed between the LPS group and I/R group (P>0.05) at 0 min after reperfusion. However, all those indexes were evidently lower in the LPS group than those in I/R group (P<0.01) at 60 min and 180 min after reperfusion. CONCLUSION:This data suggests that the protective effects induced by LPS pretreatment against hepatic I/R injury may be via down-regulation of IRAK-4 expression.     

Effect of intervention for mast cell function before reperfusion on intestinal ischemia-reperfusion-induced early liver injury in rats

AIM：To explore the effect of intervention for mast cell function before reperfusion on intestinal ischemia-reperfusion (IR)-induced early liver injury. METHODS：Adult SD rats (n=35) were randomized into 5 groups with 7 rats each: sham operation group (S group), IR group, cromolyn sodium treatment group (IR+C group, 25 mg/kg), ketotifen treatment group (IR+K group, 1 mg/kg), compound 48/80 treatment group (IR+CP group, 0.75 mg/kg). IR was induced by superior mesenteric artery occlusion for 75 min followed by 4 h of reperfusion. The agents were intravenously administered 5 min before reperfusion. The serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and histamine, and the liver levels of lactate dehydrogenase (LDH), tumor necrosis factor α (TNF-α), interleukin-8 (IL-8), malondialdehyde (MDA) and superoxide dismutase (SOD) were assessed. The liver histopathologic changes were also evaluated. RESULTS：IR resulted in severe liver injury as demonstrated by great increases in injury scores, concomitant significant increases in serum levels of AST, ALT and histamine, and liver levels of LDH, TNF-α, IL-8, and MDA, accompanied by reduced SOD activity (all P<0.05 vs S group). Treatment with cromolyn sodium or ketotifen markedly alleviated IR-mediated liver injury as confirmed by significant reduction of the above biomedical changes, whereas compound 48/80 further aggravated liver injury by dramatically enhancing the biomedical changes (all P<0.05 vs IR group). CONCLUSION：Inhibition of mast cell function before reperfusion may reduce early liver injury induced by intestinal ischemia reperfusion. Histamine, oxidative stress and inflammatory response may provide promising effects on it.     

Porcine surfactant in treatment of LPS-induced early-stage acute lung injury in rats

AIM: To evaluate the therapeutic efficacy of intratracheal instillation of porcine pulmonary surfactant (PPS) in rats with lipopolysaccharides (LPS)-induced early-stage ALI in this study.METHODS: SD rats weighing 200 g-300 g were randomly divided into 4 groups: LPS (1.5 mg·kg-1)+saline,LPS+PPS 100 mg·kg^-1,LPS+PPS 150 mg·kg^-1,LPS+PPS 200 mg·kg^-1.The PaO₂ and PaCO₂,as well as survival rate of rats were examined for 6 h after the start of PPS-instillation.Then,rats were killed and lungs were immediately removed for lung index (LI) and histological analysis.The bronchoalveolar lavage fluid (BALF) was collected for measurement of total protein (TP) contents,TNF-α level and white blood cell(WBC) numbers.RESULTS: Significantly increased PaO₂,reduced mortality rate,decreased total protein and TNF-α contents in BAL,as well as lung index and meliorated histological appearance were observed in three PPS-treated groups compared with group given saline after LPS (P<0.05).The therapeutic effect in PPS150 and PPS200 groups was better than that in PPS100 group.CONCLUSION: Intratracheal PPS instillation provides protective effect on acute lung injury in rats induced by LPS.     

Apoptosis in rat hepatocytes with ischemia/reperfusion injury

AIM: To investigate the distributive rules of apoptosis index (AI) in liver with ischemia/reperfusion (I/R) injury and evaluate the factors related to the hepatocyte apoptosis. METHODS: Sixty SD rats in specific pathogen free grade were randomly divided into three groups: control group (n=18), sham operation group (n=18) and I/R group (n=24). In I/R group, liver injury was induced by blocking blood inflow in rat liver for 20 min, then reperfusion for 22 h. Rats in the control group didnt receive any management. Rats in the sham operation group only subjected sham operation. All rat blood samples and livers were obtained for determination. Blood serum ALT, AST, TBIL, TNF-α, IFN-γ, IL-4, plasma endotoxin concentration, MDA level and SOD activity in liver were detected. Hepatic histological analysis was conduced through HE staining. Apoptosis was detected by TUNEL methods. RESULTS: Focal necrosis occurred in six rats livers in I/R group, in control group and sham group no necrosis cell was found in livers. The hepatic AI of I/R group was significantly increased compared with other groups. The AI in region under hepatic amicula was higher than that in central veins region and portal area. The necrotic regions contained apoptotic cells and AI was higher than that of other regions. Hepatic AI was significantly associated with ALT, AST, TNF-α, IFN-γ and SOD/MDA. CONCLUSION: In liver with I/R injury, the apoptotic cells in the region under hepatic amicula and the focus of necrosis are significantly higher than those in other regions, apoptotic cells and necrosis cells co-exist in the same zone. Hepatic AI may be significantly associated with ALT, AST, TNF-α and SOD/MDA.     

Protective effect of dexmedetomidine-ulinastatin combination on lipopolysaccharide-induced acute lung injury in rats

AIM：To investigate the effects of dexmedetomidine-ulinastatin combination on acute lung injury induced by lipopolysaccharide (LPS) in rats. METHODS：Male Wistar rats were randomly divided into 5 groups: saline control group (NS group) was given saline (5 mL/kg, iv) alone; LPS group (L group) was given LPS (10 mg/kg, over 10 min); dexmedetomidine+LPS group (L+D group) was treated with the additional administration of dexmedetomidine (1 μg·kg -1·h -1) immediately after LPS injection; ulinastatin+LPS group (L+U group) was treated with the addi-tional administration of ulinastatin (50 000 U/kg, ip) immediately after LPS injection; dexmedetomidine+ulinastatin+LPS group (L+D+U group) received dexmedetomidine (1 μg·kg -1·h -1) and ulinastatin (50 000 U/kg) immediately after LPS injection. The animals were sacrificed at 6 h after LPS or NS administration. Partial pressure of arterial oxygen (PaO 2), pH and base excess (BE) were measured, and the lungs were removed for evaluation of histological characteristics and determining the concentrations of TNF-α, IL-1β, macrophage inflammatory protein 2 (MIP-2), malondialdehyde (MDA), nitric oxide (NO), prostaglandin E 2 (PGE 2) and myeloperoxidase (MPO) in lung tissues, lung wet/dry weight ratio (W/D), and albumin in brochoalveolar lavage fluid (BLAF). The pulmonary expression of nuclear factor kappa B (NF-κB) p65 was evaluated by Western blotting. RESULTS：Compared with NS group, PaO 2, pH and BE was lower in L group, which was increased by treatment with dexmedetomidine-ulinastatin combination but not by dexmedetomidine or ulinastatin alone. Compared with NS group, LPS induced marked lung histological injury, which was less pronounced in the animals treated with dexmedetomidine-ulinastatin combination but not dexmedetomidine or ulinastatin alone. The levels of IL-1β, IL-6, MIP-2, MDA, NO and PGE 2 in the lung tissues increased in L group compared with NS group, which were reduced by dexmedetomidine-ulinastatin combination but not by dexmedetomidine or ulinastatin alone. The MPO activity, MDA level and W/D increased in the lung tissues in L group compared with NS group, which was reduced by dexmedetomidine-ulinastatin combination but not by dexmedetomidine or ulinastatin alone. Compared with NS group, the albumin concentration in the BLAF increased, which was reduced by dexmedetomidine-ulinastatin combination but not by dexmedetomidine or ulinastatin alone. Compared with NS group, the expression of NF-κB p65 increased in L group, which was reduced by dexmedetomidine-ulinastatin combination but not by dexmedetomidine or ulinastatin alone.CONCLUSION：Dexmedetomidine-ulinastatin combination has a protective effect on LPS-induced acute lung injury in the rats.     

Protective effects of aFGF compound solution on rat liver in hypothermic preservation

AIM: To investigate the protective effects of human acidic fibroblast growth factor (aFGF) on hypothermic preservation of rat liver. METHODS: Twenty-four Wistar rats were randomly divided into 2 groups (n=6): hypertonic citrate adenine (HCA) solution control group and experimental group (aFGF compound solution, containing 40 μg/L aFG_14-154 in HCA solution). The isolated livers were preserved in corresponding solution for 12 h or 24 h, and the content of malondialdehyde (MDA), the activity of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), and the changes of liver weight in the 2 groups were analyzed. The histopathological changes of the livers were also examined. RESULTS: Compared with HCA control group, the levels of ALT, AST and MDA and the changes of liver weight were significantly decreased in experimental group (24 h), and the changes of liver histopathology were obviously alleviated in experiment group. CONCLUSION: aFGF can reduce rat hepatic injury. The protective effects of aFGF may be related to attenuating cell swelling and improving the capacity of anti-oxidative damage in the liver.     

Minocycline inhibits rat ocular inflammation induced by lipopolysaccharide

AIM: To investigate the inhibitory effect of minocycline on the ocular inflammation induced by lipopolysaccharide in the rats. METHODS: Experimental endophthalmitis was induced in Sprague-Dawley rats by a single intravitreal injection of lipopolysaccharide (LPS of Escherichia coli, 1 μg). Minocycline (45 mg/kg) was administered intraperitoneally 12 h before and immediately after LPS injection and then every 24 h for 3 d. At 6, 12, 24, 48 and 72 h after LPS injection, eyes were graded daily for signs of clinical inflammation by slit lamp examination. The inflammatory cells were counted on histological sections. Vitreous was removed for cytokine analysis using standard enzyme-linked immunosorbent assay. Protein concentration in aqueous humor was determined.RESULTS: At all time points after LPS injection, significant clinical inhibition of ocular inflammation in the eye was observed in minocycline-treated rats. Quantitative analysis of ocular tissues revealed a significant decrease in infiltrated leukocytes ［(1 182.63±191.15) cells/eye in endophthalmitis group and (291.50±63.77) cells/eye in minocycline-treated group at 24 h postinjection, P<0.01］. Treatment with minocycline decreased tumor necrosis factor-α levels in vitreous ［(931.17±99.81)ng/L in endophthalmitis group and (353.02±71.67)ng/L in minocycline-treated group at 24 h postinjection, P<0.01］. Protein concentration in aqueous humor was decreased also, but the differences between the two groups at each time point was small and not significant (P>0.05).CONCLUSION: Minocycline treatment inhibits LPS-induced ocular inflammation with inhibition of tumor necrosis factor-α release and leukocytes infiltration. These results confirm the role of the tumor necrosis factor-α pathway and leukocytes infiltration in the pathogenesis of LPS-induced endophthalmitis, suggesting that minocycline may represent an attractive candidate for the therapeutic management of endophthalmitis.