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.     

Effect of bitter melon on liver fibrosis induced by carbon tetrachloride

AIM: To investigate the effects of bitter melon (BM) on liver fibrosis induced by CCl₄ in Wistar rats. METHODS: Healthy male Wistar rats were randomly divided into 4 groups (with 8 each): olive oil control group (group C), olive oil CCl₄ model group (group M), CCl₄+BM at low concentration (BM 100 g/kg, group BM-L), CCl₄+ BM at high concentration (BM 200 g/kg, group BM-H). All rats except those in group C were subcutaneously injected with CCl₄ twice a week for 8 weeks to induce liver fibrosis. After injection of CCl₄ for 8 weeks, all rats were sacrificed and the samples of blood and livers were collected. The weight ratio of liver to body was measured. The serum level of MDA and the activity of SOD were tested. The contents of total protein and albumin, the activity of GSH-Px, the content of hydroxyproline and the activity of monoamine oxidase in the liver homogenate were determined. Hepatic inflammation and collagen deposition were observed under microscope with Masson staining. RESULTS: In the rats treated with BM, the weight ratio of liver to body, the serum level of MDA, the content of hydroxyproline and the activity of monoamine oxidase in the liver homogenate were lower than those in group M (P<0.01). The serum activity of SOD, the contents of total protein and albumin, and the activity of GSH-Px in the liver homogenate were enhanced (P<0.01). The livers of the model rats had remarkable inflammatory necrosis, collagen accumulation and fibrosis. The rats in BM-treated group showed slighter hepatic injury and collagen deposition, and the liver functions were much better than those in the model group. High dose of BM showed more obvious liver-protective effects. CONCLUSION: BM attenuates liver fibrosis by its antioxidant effect and the mechanisms of reducing hydroxyproline content and monoamine oxidase activity.     

Changes of liver sinusoidal endothelial cells and basement membrane in early stage of liver fibrosis induced by carbon tetrachloride in rats

AIM：To explore the development of hepatic sinusoidal capillarization in the early stage of liver fibrosis induced by carbon tetrachloride (CCl4) in rats. METHODS：Clean SD rats were randomly divided into normal control group (group N, n=6) and liver fibrotic model group (group M, n=32). The rats in group N were intraperitoneal injected with saline and the rats in group M were intraperitoneal injected with CCl4 (2 mL/kg, twice a week for 4 weeks). At the end of the 3rd day and the 1st, 2nd and 4th weeks, all rats were killed and then the samples were collected. The pathological changes in the livers were observed by HE staining and Masson straining. The development of hepatic sinusoidal capillarization was observed by transmission electron microscopy (TEM) and immunohistochemical staining. The cell surface expression of vascular endothelium-associated marker CD31, collagen type Ⅳ (Col IV) and laminin (LN) was determined. RESULTS：HE and Masson staining showed the formation of liver fibrosis after treatment with CCl4 for 4 weeks. TEM showed that the fenestrate diameter of liver sinusoidal endothelial cells (LSECs) grew down, the fenestrate numbers of LSECs were decreased along with the development of liver fibrosis, and the consecutive basement membrane was formed at the end of the experiment. The expression of CD31 was significantly increased along with the development of defenestration, and the expression of Col IV and LN was significantly increased after the treatment with CCl4 for 2 weeks and 4 weeks, respectively. CONCLUSION：The typical hepatic sinusoidal capillarization was detected in the early stage of liver fibrosis, and the deposition of LN in the liver sinusoidal walls was the mainly factor of formation of the consecutive basement membrane.     

Hepatoprotective effect of Paecilomyces tenuipes polysaccharides on carbon tetrachloride -induced liver injury

AIM:To investigate the protective effect of crude Paecilomyces tenuipes polysaccharides (cPtPs) and Paecilomyces tenuipes polysaccharides (PtPs) in a rat model of acute hepatic necrosis induced by carbon tetrachloride (CCl4). METHODS:Wistar rats were divided into four groups (control group, CCl4 group, cPtPs+CCl4 group and PtPs +CCl4 group), the four groups were given intragastrically with normal saline, cPtPs and PtPs for 15 d, respectively. In the last two days, these groups except control group were injected peritoneally with CCl4. Serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL), direct bilirubin (DBIL) and indirect bilirubin (IBIL) were detected by automatic biochemistry analyzer. Pathological changes of hepatic tissue were assessed by hematoxylin-eosine (HE) staining. The levels of superoxide dismutase (SOD) and malondialdehyde (MDA) in liver homogenate were analyzed using xanthinoxidase and thio-barbituric acid, respectively. The concentration of Ca2+ in hepatocyte mitochondria was determined by colorimetric method. The expression of α-smooth muscle actin (α-SMA) was examined in hepatic tissue by immunohistochemical method. RESULTS:Compared with control group, serum levels of ALT, AST, TBIL, DBIL and IBIL in CCl4 group increased significantly, denaturation and necrosis implicated to the whole hepatic lobules. Compared with CCl4 group, serum levels of ALT, AST, TBIL, DBIL and IBIL in PtPs +CCl4 group decreased significantly, denaturation and necrosis located in the third region of hepatic lobules, the level of SOD increased and MDA decreased (P<0.05) in endochylema. Concentration of Ca2+ in mitochondria decreased in PtPs +CCl4 group and cPtPs +CCl4 group (P<0.05, P<0.01, respectively). Expression of α-SMA was found little in PtPs+CCl4 group. CONCLUSION:PtPs, the effect is better than cPtPs, lessens CCl4-induced hepatonecrosis significantly. The role may be related with anti-lipid peroxidation.     

Ethanol extract from Cortex Albiziae attenuates mouse acute liver injury induced by carbon tetrachloride via modulation of NF-κB pathway

AIM:To investigate the protective effect of ethanol extract from Cortex Albiziae on acute liver injury, and to explore its possible mechanism. METHODS:Acute liver injury in mice was induced by single intraperitoneal injection of 25% carbon tetrachloride (olive oil solubilization). The effective parts of ethanol extract from Cortex Albizziae against acute liver injury were screened. The pathological changes of the liver tissues were examined by pathological sections with HE staining. The activity of total superoxide dismutase (T-SOD) and the content of malondialdehyde (MDA) of the liver tissues were detected, the serum levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were mea-sured by ELISA, and the protein expression levels of NF-κB p65, Bcl-2 and Bax in the liver cells of the mice in each group were determined by Western blot. RESULTS:Compared with model group, the serum levels of AST and ALT in low-dose n-butanol phase of ethanol extract from Cortex Albiziae (AB-L, 4 mg·kg^-1·d^-1) group and high-dose n-butanol phase of ethanol extract from Cortex Albiziae (AB-H, 8 mg·kg^-1·d^-1) group were significantly decreased. The necrosis extent and degree of the hepatocytes and infiltration of inflammatory cells were significantly lower than that in model group. Compared with model group, the serum levels of TNF-α and IL-6 in AB-H group and AB-L group were significantly decreased (P<0.05). The protein level of NF-κB p65 in the nuclei of mouse liver cells in AB-H group and AB-L group were also decreased significantly (P<0.05). Compared with model group, the protein expression of Bax was decreased, the protein expression of Bcl-2 was increased, and the Bcl-2/Bax ratio was increased in AB-L group and AB-H group. CONCLUSION:The n-butanol phase of ethanol extract from Cortex Albiziae may protect the liver by reducing the activation of NF-κB p65, inhibiting the excessive release of inflammatory cytokines IL-6 and TNF-α, and decreasing hepatocyte apoptosis via regulating Bcl-2 and Bax expression.     

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.     

Activation of farnesoid X receptor attenuates liver injury in MRL/lpr lupus mice

AIM: To observe how the activation of farnesoid X receptor (FXR) influences liver function in MRL/lpr lupus mice. METHODS: The expression of FXR at mRNA and protein levels was determined in the liver specimens of MRL/lpr mouse model.MRL/lpr mice and the control BALB/c mice received concanavalin A (ConA) to induce liver injury. The FXR agonist chenodeoxycholic acid (CDCA) was administered to MRL/lpr mice. Blood samples were taken at the time points of 0 h, 4 h, 8 h, 12 h, 16 h and 24 h after ConA injection for the detection of serum ALT, AST, IFN-γ, TNF-α and IL-6. RESULTS: FXR was down-regulated in the liver specimens of MRL/lpr mice. MRL/lpr mice were more susceptible to ConA than BALB/c mice to induce significantly higher levels of aminotransferases and inflammatory cytokines. Activation of FXR by CDCA significantly reduced the levels of aminotransferases and inflammatory cytokines IFN-γ, TNF-α and IL-6 caused by ConA injection in MRL/lpr mice. CONCLUSION: FXR activation ameliorates liver injury and suppresses the production of inflammatory cytokines, indicating that FXR protects the liver functions in lupus mice.     

Protective effect of emodin on lung injury induced by hepatic fibrosis in rats

AIM:To explore the protective effect of emodin on lung injury induced by hepatic fibrosis in rats. METHODS:The hepatic fibrosis rat model was established with multiple pathogenic factors (CCl 4, ethanol, high fat, high cholesterol and low choline) and treated with different doses (20 mg/kg and 40 mg/kg) of emodin for 4 weeks. The hepatic index was measured. The biochemical indexes, endotoxin, homocysteine, albumin, aspartate aminotransferase,alanine aminotransferase, total bilirubin, total cholesterol and triglyceride, and hepatic fibrosis indexes, hyaluronic acid, laminin, collagen IV and procollagen Ⅲ, were detected. The histopathological changes of the liver were observed. The pulmonary index was determined. The histopathological changes of the lungs were observed. The levels of tumor necrosis factor α (TNF-α), malondialdehyde (MDA), nitric oxide (NO) and peroxynitrite (ONOO-) in the lung tissues were analyzed. RESULTS:The rat hepatic fibrosis model was successfully established. In model group, lung edema and inflammation occurred, and the pulmonary index and the levels of TNF-α, MDA, NO and ONOO- in the lung tissues were increased significantly. In emodin treatment groups, the pulmonary indexes were lower than that in model group. The pathological injury of the lung tissues was alleviated. The levels of TNF-α, MDA, NO and ONOO- in the lung tissues were decreased. CONCLUSION:Emodin has a protective effect on lung injury induced by hepatic fibrosis in rats.     

Up-regulation of CD36 induced by casein-induced inflammation promotes renal injury in mice

AIM: To investigate the role of CD36 in casein-induced mouse renal injury.METHODS: Eight-week-old male C57BL/6J mice and CD36 knockout (CD36KO) mice were randomly divided into C57BL/6J saline injection group, C57BL/6J casein injection group and CD36KO casein injection group (n=8 in each group). After 14 weeks of treatment with high-fat diet, the mouse serum, 24 h urine and kidney tissue samples were collected for analysis. The serum content of tumor necrosis factor-α (TNF-α) was measured by ELISA. The renal function markers in the serum and urine were determined by an automatic biochemical analyzer. The pathological changes of the kidney were observed by HE staining and Masson staining. The expression of CD36 and cytokines/chemokines (TNF-α, IL-6 and MCP-1) at mRNA and protein levels in the renal tissues were determined by real-time PCR and Western blot. The content of tissue hydrogen peroxide (H₂O₂) was measured by a commercial kit. The protein levels of Nrf2 and TGF-β1 in the renal tissues were measured by immunohistochemical staining.RESULTS: Compared with saline injection group, casein injection increased the level of TNF-α in the serum and in the kidney tissues of C57BL/6J mice (P<0.05), suggesting that casein injection successfully induced chronic inflammation in C57BL/6J mice. Casein injection also promoted the protein expression of CD36 and TGF-β1 in the renal tissues of the C57BL/6J mice, accompanied with glomerular sclerosis, proteinuria, increased serum creatinine content, increased H₂O₂ content, and decreased Nrf2 protein level and the ability of antioxidant in the kidneys (P<0.05). Furthermore, CD36 deficiency protected the mice from casein-induced renal injury, as evidenced by improved kidney pathological changes and decreased proteinuria. The content of H₂O₂ in the kidneys of casein-treated CD36 knockout mice was also lower than that in casein-treated C57BL/6J mice.CONCLUSION: Inflammatory responses promote the oxidative stress and renal injury in a CD36-dependent manner.     

Rebamipide repairs injury of small intestinal epithelial barrier induced by aspirin in mice

AIM: To investigate whether rebamipide repairs the small intestinal epithelial barrier in aspirin-induced small intestinal injury (SⅡ) in mice and its mechanism.METHODS: Small intestinal injury was induced by aspirin (200 mg·kg^-1·d^-1 for 5 d) in BALB/c mice. Based on the treatment with aspirin and/or rebamipide (320 mg·kg^-1·d^-1), the mice were divided into 4 groups (n=18 in each group). The living mice in each group (n=6) were sacrificed via cervical dislocation method at day 0, day 5, and day 10. The structure and function of intestinal barrier and the levels of the signaling pathway factors were measured by transmission electron microscopy, immunohistochemistry, qPCR, and Western blot.RESULTS: Tight junctions between intestinal epithelial cells improved significantly after reba-mipide treatment. The expression of ZO-1 and occludin in the injured small intestine showed a gradually increasing trend after rebamipide administration (P<0.05). There was a decreased trend of D-lactate level in rebamipide-treated SⅡ mice (P<0.05). The expression of cyolooxygenase-2 (COX-2), β-catenin, and c-Myc, and prostaglandin E₂ concentration in small intestinal tissues were significantly increased in rebamipide treatment group (P<0.05). However, down-regulated COX-1 expression in the SⅡ mice was sustained at a low level after rebamipide administration.CONCLUSION: Rebami-pide repairs the injury of small intestinal mucosa and improves the structure and function of small intestinal barrier in aspirin-induced SⅡ mice by up-regulating the expression of COX-2.     

柳松菇对急性四氯化碳肝损伤的防护作用

利用柳松菇水提物对小鼠急性四氯化碳肝损伤的防护作用进行了研究。结果证明，柳松菇对急性CCl4肝损伤有良好的保护作用。     

The reversal effects of recombinant human augmenter of liver regeneration in immunocomplex induced liver fibrosis

AIM: To observe the anti-fibrosis activity of recombinant human augmenter of liver regeneration (hALR). METHODS: Albumin induced rat model of liver fibrosis was established and hALR was given peritoneally after the model production. Serum concentration of alanine aminotransferase(ALT),aspartic aminotransferase(AST),lactate dehydrogenase(LDH), hepatic collagen contents and pathological examination were selected as observing parameter. RESULTS: Recombinant human augmenter of liver regeneration (hALR) could decrease ALT, AST, LDH concentration of fibrotic rats. The measurements of hepatic collagen contents showed that hepatic collagen contents in hALR treatment group was much lower than those of model group and negative control group. Pathological examination also indicated that the degree of liver fibrosis in hALR treatment group was attenuated in comparison with those of model group and negative control group. CONCLUSION: Recombinant human augmenter of liver regeneration (hALR) had reversal effects on immunocomplex induced rat liver fibrosis.control group. CONCLUSION: Recombinant human augmenter of liver regeneration (hALR) had reversal effects on immunocomplex induced rat liver fibrosis.     

Characteristics in the secretion of liver TNF-α induced by infusion of LPS into veins in goats

AIM: To study the role of liver in immune regulation in experimental endotoxemia. METHODS: 17 castrated male goats were subjected to simultaneously installing catheters in jugular, hepatic and portal veins by surgery. Four days later, lipopolysaccharide (LPS) was infused in term of three groups as followings: In group ①, LPS of 20 EU (endotoxin unit, EU)·kg^-1 was infused into portal vein; In group ②, LPS of 20 EU·kg^-1 was infused into jugular vein and LPS of 1 500 EU·kg^-1 infused into jugular vein in group ③. Before and after infusion, blood samples were collected from the three veins through the catheters for 8 h.The plasma levels of TNF-α were measured by RIA. RESULTS: In group ①, the plasma TNF-α levels of hepatic and portal vein rose to peak value at 5 h, but that of the jugular vein did not changed. In group ②, the plasma TNF-α levels in hepatic vein rose to peak value at 3 h. The TNF-α levels of jugular vein rose to peak value at 1 h and the one in portal vein enhanced continuously between 0-8 h. In group ③, the plasma TNF-α levels in jugular, hepatic and portal vein rose to significant peaks at 1 h simultaneously. CONCLUSION: During experimental endotoxemia,liver showed different dynamic characteristics in TNF-α secretion according to the pathway and doses of LPS delivery.     

Functional analysis of differential microRNA expression profile in mouse fibrotic liver tissues induced by CCl4

AIM:To discover the expression profile of microRNAs (miRNAs) in mouse fibrotic liver tissues induced by carbon tetrachloride (CCl4), and to investigate the functions of these differential miRNAs based on the gene ontology (GO) analysis and KEGG Pathway analysis. METHODS:The mice were randomly divided into normal group and model group. Liver fibrosis was induced by subcutaneous injection of CCl4. miRNA expression profile of the liver tissues was assayed by a mouse miRNA microarray (Agilent 12.0). The differential expression of miRNAs between the normal and model mice was screened, and GO analysis and KEGG Pathway analysis were performed to determine the functions of these differential miRNAs. RESULTS:Thirty-nine miRNAs with differential expression were discovered in the model mice compared with the normal mice, among which 23 were up-regulated and 16 were down-regulated. GO analysis and KEGG Pathway analysis indicated that most pathological processes of liver fibrosis regulated by miRNAs included cell proliferation and activation, cell apoptosis, cell cycle, cell adhesion, inflammatory reaction, cell migration, transforming growth factor β (TGF-β) signaling pathway, Wnt signaling pathway and proteometabolism process. GO analysis revealed that the key up-regulated miRNAs were mmu-miR-322, mmu-miR-15b, mmu-miR-195, mmu-miR-200b and mmu-miR-214, and the key down-regulated miRNAs were mmu-miR-16, mmu-miR-130a, mmu-miR-101b, mmu-miR-30a and mmu-miR-30e. Analyzing the target genes screened out by GO analysis and Pathway analysis simultaneously, we found that the key up-regulated miRNAs included mmu-miR-200b, mmu-miR-322, mmu-miR-106b, mmu-miR-23a and mmu-miR-15b, and the key down-regulated miRNAs included mmu-miR-16, mmu-miR-30e, mmu-miR-30c, mmu-miR-30a and mmu-miR-130a. CONCLUSION: Differential expression of miRNAs is discovered in mouse fibrotic liver tissues induced by CCl4 compared with the normal liver tissues. Most of the pathological processes involved in liver fibrosis may be regulated by miRNA, such as cell proliferation and activation, cell adhesion and apoptosis, cell migration and differentiation, metabolism, TGF-β receptor signaling pathway and so on.     

Role of carbon monoxide in CCK-8 ameliorating acute lung injury induced by LPS

AIM: To study the role of carbon monoxide (CO) in the mechanism of cholecystokinin-octapeptide (CCK-8) for attenuation of acute lung injury (ALI) induced by lipopolysaccharide (LPS). METHODS: Fifty-six adult male rats were randomly divided into seven groups: control group, LPS group, LPS+ZnPP (a specific inhibitor of HO-1) group, LPS+Hemin (Hm， CO donor) group, CCK-8+LPS group, CCK-8+LPS+ZnPP group and CCK-8 group (n=8 for each). Bronchoalveolar lavage was performed 2 h, 6 h and 12 h respectively after treatments. The numbers of polymorphonuclear leukocytes (PMN) in bronchoalveolar lavage fluid (BALF) was detected. The mortality of rats and the structure of lung tissues were observed. MDA and CO contents in lung tissues were also measured. RESULTS: The mortalities of rats were both zero 2 h and 6 h after agent administration. The mortality of rats was higher than control group 12 h after LPS administration. The mortality of rats in LPS+Hm and CCK-8+LPS group were lower than that in LPS group, and its in LPS+ZnPP and CCK-8+LPS+ZnPP group were lower than that in LPS and CCK-8+LPS group, respectively. Lung injury was observed in LPS group. At the same time the number of PMN, MDA and CO content were higher than those in control group. The degree of lung injury, PMN numbers and MDA content were lower, while CO content in LPS+Hm and CCK-8+LPS group were higher than those in LPS group. However, the degree of lung injury, PMN number and MDA content were higher, CO content were lower in LPS+ZnPP and CCK-8+LPS+ZnPP group than those in LPS and CCK-8+LPS group, respectively. CONCLUSION: CCK-8 attenuates the LPS-induced acute lunginjury by means of anti-oxidation and inhibition of PMN aggregation, which are both mediated by CO.     

Inhibitory effects of Angelicae sinensis and sodium ferulate on acute inflammatory liver injury and expression of ICAM-1 and E-selectin in mice

AIM: To explore the effects of Angelicae sinensis preparation and sodium ferulate on inflammatory liver injury induced by lipopolysaccharide (LPS) and their molecule mechanism. METHODS: ICR mice were divided into five groups: Angelicae sinensis group, sodium ferulate group, dexamethasone group, inflammation control group and normal group. The model of inflammatory injury in mice was set up by tail vein injection with the bacillus calmette-guerin (BCG) and LPS respectively prior and posterior to administration of those tested drugs. The tested drugs (the preparations of Angelica sinensis, sodium ferulate and dexamethasone) and normal saline were given respectively to the corresponding group. The pathological observation of the liver tissues in mice was made for the intensity of inflammatory liver injuries. The immunohistochemical detection and comparison were performed for the expression of ICAM-1 and E-selectin proteins in the liver tissues in those mice. RESULTS: The intensities of liver inflammatory injuries in mice from drug-treated groups were obviously lighter than that from the inflammation control group (P<0.01). The expressions of ICAM-1 and E-selectin proteins in the liver tissues of mice from the inflammation control group were not only significantly higher than that in normal control, but also obviously higher than that from drug-treated groups (P<0.01). CONCLUSIONS: Angelica sinensis and sodium ferulate alleviate inflammatory liver injury induced by injection of LPS. Their suppressive effects on inflammatory liver injury may be associated with the decrease in the expression of ICAM-1 and E-selectin proteins.     

Role of hydrogen sulfide in alleviation of liver injury by mesenteric lymph drainage in hemorrhagic shock rats

AIM：To investigate the role of hydrogen sulfide (H2S) in alleviation of liver injury by mesenteric lymph drainage in hemorrhagic shock rats. METHODS：A hemorrhagic shock model was established in male Wistar rats. DL-propargylglycine (PPG), an inhibitor of cystathionine γ-lyase (CSE) which is a synthase of H2S, or sodium hydrosulfide (NaHS), a donor of H2S, was administered to the hemorrhagic shock rats with mesenteric lymph drainage. The rats were randomly divided into sham, shock, shock+drainage, shock+drainage+PPG (45 mg/kg, ip, 0.5 h before hemorrhage) and shock+drainage+NaHS (28 μmol/kg, ip, 0.5 h before hemorrhage) groups. Fluid resuscitation was performed 1 h after hypotension, and then mesenteric lymph was drained in the rats of shock+drainage, shock+drainage+PPG and shock+drainage+NaHS groups for 3 h. The hepatic histomorphology was observed. The biochemical indexes of hepatic function in plasma, and H2S, CSE, Toll-like receptor 4 (TLR4), interleukin (IL)-10, IL-12 and tumor necrosis factor α (TNF-α) in hepatic homogenate were also examined. RESULTS：The levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and total bile acid (TBA) in plasma, and H2S, CSE, TLR4, IL-10, IL-12 and TNF-α in hepatic homogenate in shock group were significantly higher than those in sham group. Mesenteric lymph drainage obviously reduced these indexes in shock rats, except for TLR4. PPG further decreased these indexes except for CSE, while NaHS increased these indexes except for TBA and CSE. Morphological observation showed that liver injury appeared in the rats from shock and shock+drainage+NaHS groups, and there was nearly normal hepatic structure in the rats from sham, shock+drainage and shock+drainage+PPG groups. CONCLUSION：The mechanism of mesenteric lymph drainage alleviating liver injury in hemorrhagic shock rats is related to reducing the production of H2S and alleviating the H2S-mediated inflammation.     

Exogenous carbon monoxide protects against the lung injury induced by ischemia-reperfusion of hind limbs in rats

AIM: To study the mechanism of protective effect of exogenous carbon monoxide (CO) in the lung injury induced by ischemia-reperfusion (IR) of hind limbs in rats. METHODS: Thirty-two SD rats were randomly divided into 4 groups: control, control+CO, IR and IR+CO. A rat model of ischemia in hind limbs and the reperfusion lung injury was made. The rats in IR+CO and control+CO groups were exposed to air containing 2.5×10-8 CO for 1 h before reperfusion or the corresponding control time point, while the other two groups were exposed to the routine air. The lung tissue structure, polymorphonuclear leukocyte (PMN) count, wet-to-dry weight ratio (W/D), malondialdehyde (MDA) content and the animal survival rate were observed. The carboxyhemoglobin (COHb) levels in artery blood were detected with CO-oximeter and the expression of intercellular adhesion molecule-1 (ICAM-1) in the lung was detected by Western blotting. RESULTS: Compared to control, the animal mortality, lung PMNs number, W/D, MDA content and ICAM-1 expression were all significantly increased in IR group. Compared with the IR group, the blood COHb level was significantly increased and the animal mortality, lung PMNs number, W/D, MDA content and ICAM-1 expression were all significantly decreased in IR+CO group. CONCLUSION: These data suggest that exogenous CO attenuate limb IR-induced lung injury by down-regulatiny ICAM-1 expression and suppressing PMN sequestration in the lung following limb IR in rats.     

Effect of betaine on liver injuries in aging db/db mice induced by lipids

AIM: To evaluate the effect of betaine on lipid metabolism disorder in inherited db/db mice with long-term nonalcoholic fatty liver disease (NAFLD).METHODS: Experimental NAFLD models were established by feeding the db/db mice with high-fat diet. Fifty 7-month-old db/db mice were randomly divided into 5 groups: the mice in low, medium and high dose groups were given betaine by intragastric administration at doses of 200 mg/kg, 400 mg/kg and 800 mg/kg for 6 weeks, respectively,while the mice in saline control group and positive drug group were given normal saline and positive control drug,respctively. All the animals were killed, and serum alanine aminotransferase (ALT), triglyceride (TG), total cholesterol (TC), low-density lipoprotein (LDL) and glucose tolerance were detected. The pathological changes of the liver tissues were also observed.RESULTS: Betaine significantly decreased the levels of ALT, TC and LDL (P<0.05 or P<0.01). The pathological changes of the liver tissues indicated that the content of lipid in the hepatocytes of betaine treatment groups was less than that in saline control group.CONCLUSION: Betaine significantly improves the lipid metabolism and the liver function in the aging db/db mice, and reduces the accumulation of lipid in the hepatocytes.     

Pretreatment of hypertonic saline attenuates the hepatic ischemia reperfusion injury induced by neutrophils

AIM: To explore the effect of the pretreatment of hypertonic saline (HTS) in hepatic ischemia reperfusion (I/R) injury.METHODS: The rats were divided into sham group (sham group), ischemia reperfusion group (IR group) and pretreatment of hypertonic saline group (HTS group). Partial hepatic ischemia reperfusion model was used. The rats were sacrificed at the time of 1 h, 3 h, 6 h, 12 h and 24 h after reperfusion in each group, respectively. Blood samples were obtained to examine ALT. The expression of the CD11b/CD18 (Mac-1) on the neutrophils was analyzed by flow cytometry. RT-PCR and Western blotting were used to examine the expression of intercellular adhesion molecule-1 (ICAM-1) in livers and chromatometry was performed to detect the activity of myeloperoxidase (MPO) in livers. The morphology of hepatocytes and the structure of sinusoid were observed by histological examinations. RESULTS: ① HTS pretreatment decreased the level of ALT at the time points of 3 h, 6 h and 12 h after reperfusion (P<0.05). ② Mac-1 expression in HTS group was lower at 6 h and 12 h after reperfusion compared with IR group (P<0.05). ③ MPO activity in HTS group was lower at 6 h, 12 h and 24 h compared with IR group (P<0.05). ④ RT-PCR and Western blotting analysis indicated that the pretreatment of HTS inhibited the expression of ICAM-1 in livers after reperfusion. ⑤ Moderate hepatocyte swelling and few neutrophil infiltration were observed in HTS group.CONCLUSION: Pretreatment with HTS has the effect on hepatic ischemia reperfusion injury by inhibiting the expression of Mac-1 on circulating neutrophils and the expression of ICAM-1 in the liver.