OBJECTIVETo investigate the role of excretory/secretory antigens from Clonorchis sinensis (CsESAs) in hepatic fibrosis induced by C. sinensis infection in rats and explore the possible mechanism.

METHODSCsESAs was collected from adult C. sinensis cultured in sterile condition for 12 h and injected intraperitoneally in Wistar rats. Masson staining was used to observe the changes in the hepatic collagen fiber after the injection. HE staining and immunofluorescence staining were performed to detect the expression of alpha-smooth muscle actin (alpha-SMA) to examine the proliferation and the activity of hepatic stellate cells. The specific antibody titer of CsESAs was determined using enzyme-linked immunosorbent assay to investigate the role of the antigen-antibody complex in the development of hepatic fibrosis.

RESULTSAfter intraperitoneal injection of CsESAs, obvious hepatic fibrosis and hepatic stellate cell proliferation and activation were observed in the rat livers. The severity of the hepatic fibrosis was associated with the dose of CsESAs injected, whereas the titer of the specific antibody against CsESAs showed no direct relation to the hepatic fibrosis.

CONCLUSIONIntraperitoneal injection of CsESAs can cause hepatic stellate cell activation and hepatic fibrosis in rats, but the antigen-antibody complex does not seem to play the key role in the activation of the hepatic stellate cells.

Actins ; metabolism ; Animals ; Antigens, Helminth ; immunology ; Clonorchiasis ; parasitology ; Clonorchis sinensis ; immunology ; pathogenicity ; Hepatic Stellate Cells ; pathology ; Liver Cirrhosis ; immunology ; parasitology ; Male ; Rats ; Rats, Wistar

This study examined endogenous cannabinoid (ECB)-anandamide (AEA) and its cannabinoid receptors (CBR) in mice liver with the development of schistosoma japonicum. Mice were infected with schistosoma by means of pasting the cercaria onto their abdomens. Liver fibrosis was pathologically confirmed nine weeks after the infection. High performance liquid chromatography (HPLC) was employed to determine the concentration of AEA in the plasma of mice. Immunofluorescence was used to detect the expression of CBR1 and CBR2 in liver tissue. Morphological examination showed typical pathological changes, with worm tubercles of schistosoma deposited in the liver tissue, fibrosis around the worm tubercles and infiltration or soakage of inflammatory cells. Also, CBR1 and CBR2 were present in hepatocytes and hepatic sinusoids of the two groups, but they were obviously enhanced in the schistosoma-infected mice. However, the average optical density of CBR1 in the negative control and fibrosis group was 13.28+/-7.32 and 30.55+/-7.78, and CBR2 were 28.13+/-6.42 and 52.29+/-4.24 (P<0.05). The levels of AEA in the fibrosis group were significantly increased as compared with those of the control group. The concentrations of AEA were (0.37+/-0.07) and (5.67+/-1.34) ng/mL (P<0.05). It is concluded that the expression of endocannabinoids AEA and its cannabinoid receptor CBR were significantly increased in schistosoma-infected mice. Endogenous endocannabinoids may be involved in the development of schistosoma-induced liver fibrosis.
Arachidonic Acids/*metabolism ; Endocannabinoids/*metabolism ; Liver Cirrhosis/etiology ; Liver Cirrhosis/*metabolism ; Liver Cirrhosis/parasitology ; Polyunsaturated Alkamides/*metabolism ; Random Allocation ; Receptor, Cannabinoid, CB1/*metabolism ; Receptor, Cannabinoid, CB2/*metabolism ; Schistosomiasis japonica/*complications ; Schistosomiasis japonica/metabolism

Animals ; Female ; Liver Cirrhosis, Experimental ; drug therapy ; parasitology ; Mice ; Mice, Inbred Strains ; Pentoxifylline ; therapeutic use ; Schistosomiasis japonica ; drug therapy

BACKGROUND AND OBJECTIVES: Placenta and blood that remained in the umbilical cord is routinely available as a discarded tissue after deliveries and it is free of any legal, moral, ethical or religious objections, providing a high number of multipotent CD34+ progenitor and stem cells. Using ex vivo isolated CD34+ cells from human umbilical cord blood (hUCB) have emerged as promising candidates to treat various diseases, including exogenous pathogenic infections. We have expanded to build a rational approach to study the effect of CD34+ cells after damaged liver tissues by the devastating human parasitic flatworm Schistosoma mansoni. METHODS AND RESULTS: Experimental studies were conducted in the Department of Zoology, Faculty of Science and Departments of Parasitology and Physiology, Faculty of Medicine, SCU, Egypt. We have studied the impact of ex vivo preparation of CD34+ cells from hUCB on S. mansoni-induced liver fibrosis de novo, and treated for shorter and longer periods in vivo. Ova count, ALT and albumin were measured at specific time interval and histopathological examination of liver was conducted to confirm the biochemical results. The data obtained were statistically analyzed by ANOVA between groups. It was found that the administration of CD34+ cells have modestly reduced liver damage; reduced the S. mansoni infection associated elevation in serum levels of ALT; significantly improved serum levels of albumin and reduced egg granuloma diameter in the livers. CONCLUSIONS: We demonstrated that CD34+ cells can markedly ameliorated liver fibrosis in vivo and may be beneficial for therapy to recover organ structure and/or function of S. mansoni-infected mice.
Animals ; Egypt ; Fetal Blood ; Fibrosis ; Granuloma ; Humans ; Liver ; Liver Cirrhosis ; Mice* ; Ovum ; Parasitology ; Physiology ; Placenta ; Platyhelminths ; Schistosoma mansoni* ; Stem Cells ; Umbilical Cord ; Zoology

Animals ; Cannabinoid Receptor Modulators ; Liver ; metabolism ; Liver Cirrhosis, Experimental ; metabolism ; parasitology ; Male ; Mice ; Mice, Inbred Strains ; Receptor, Cannabinoid, CB1 ; metabolism ; Schistosoma mansoni ; Schistosomiasis mansoni ; metabolism

OBJECTIVETo investigate the effects of sympathetic neurotransmitters and adrenergic receptors on liver fibrosis in murine schistosomiasis.

METHODSMice were infestated with schistosoma by means of pasting cercariae on their abdomens. Thirty mice were randomly divided into a control group and a model group. Hematoxylin eosin and Van Gieson staining were used to view the histopathology of their livers. Immunofluorescence histochemistry and laser scanning confocal fluorescence microscopy were used to measure the a1A and beta2 adrenergic receptors in livers of the two groups of mice. High performance liquid chromatography-electrochemical detector (HPLC-ECD) was used to determine the concentration of norepinephrine (NE) and dopamine (DA) in the plasma of the mice.

RESULTSImmunofluorescence histochemistry showed that a1A and beta2 receptors were present in hepatocytes and hepatic sinusoids of the livers of the mice of the two groups, but there were many more in the livers of the schistosoma infected mice (t=-2.888; t=-6.648) (P<0.05). The results of HPLC-ECD showed that the levels of NE and DA in the model group were higher than those of the control group (t=-3.372; t=-4.428) (P<0.05).

CONCLUSIONSympathetic neurotransmitters and adrenergic receptors may participate in liver fibrogenesis in mice infected with schistosoma.

Animals ; Dopamine ; blood ; Liver ; pathology ; Liver Cirrhosis ; metabolism ; parasitology ; pathology ; Male ; Mice ; Mice, Inbred Strains ; Neurotransmitter Agents ; blood ; Norepinephrine ; blood ; Receptors, Adrenergic ; blood ; Schistosomiasis ; metabolism

Chronic Opisthorchis viverrini-induced hepatobiliary disease is associated with significant leukocyte infiltration, including activated macrophages; however, the polarization of infiltrating macrophages remains to be fully characterized. In this study, we characterized macrophage polarization and phenotype in chronic O. viverrini-induced hepatobiliary disease in humans and hamsters using gene expression and histochemical analysis. Chronic O. viverrini infection and associated hepatobiliary diseases were associated with iron loaded M2-like macrophages in both humans and hamsters. This study provides suggestive evidence that iron loaded M2-like macrophages promote hepatobiliary disease in chronic O. viverrini infection.
Animals ; Cricetinae ; Gene Expression Profiling ; Histocytochemistry ; Humans ; Immunohistochemistry ; Iron/*metabolism ; Liver Cirrhosis/*parasitology/*pathology ; Macrophages/*immunology/metabolism ; Mesocricetus ; Opisthorchiasis/*complications/*pathology ; Opisthorchis/*isolation & purification

Animals ; Female ; Interferon-alpha ; therapeutic use ; Interferon-gamma ; therapeutic use ; Liver Cirrhosis ; drug therapy ; parasitology ; Mice ; Mice, Inbred ICR ; Schistosoma japonicum ; Schistosomiasis japonica ; complications

OBJECTIVETo investigate the action of nitric oxide (NO) on testicular dysfunction in cirrhotic rats.

METHODSCirrhotic rats were induced by bile duct ligation (BDL). Concentration of NO in the serum and homogenates of the testicular tissue in biliary cirrhotic rats, L-NAME rats, and sham operated rats were measured by assay of nitrate reductase. Concentrations of testosterone in the serum of 3 groups were measured by radioimmunoassay. Sperm density and percent of motive sperm in the epididymis of the rats were determined.

RESULTSConcentrations of NO in the serum and homogenates of the testicular tissue of cirrhotic rats were significantly greater than those of sham operated rats (4.165 micromol/L 1.162 micromol/L, and 1.305 micromol/g 0.087 micromol/g vs 0.535 micromol/L 0.237 micromol/L and 0.720 micromol/g 0.063 micromol/g). Concentrations of testosterone in the serum, the sperm density and percent of motive sperm in the epididymis were significantly lower in cirrhotic rats than sham operated rats (0.049mug/L 0.020 microgram/L, 16.46% 4.84%, and 86.89 10(6)/ml 33.17 10(6)/ml vs 2.680 microgram/L 0.403 microgram/L, 62.45% 9.21%, and 299.43 10(6)/ml 53.85 10(6)/ml). By contrast, the administration of a low dose of L-NAME (0.5 mg/kg per day) for one week to cirrhotic rats was associated with a significant reduction in concentration of NO (1.975 micromol/L 0.406 micromol/L and 0.950 micromol/g 0.057 micromol/g) and a significant increase in concentration of testosterone in the serum, the sperm density and percent of motive sperm in the epididymis (0.993 microgram/L 0.179 microgram/L, 33.85% 4.93%, and 188.94 10(6)/ml 38.34 10(6)/ml).

CONCLUSIONSNO is associated with testicular dysfunction in cirrhosis. The testicular dysfunction induced by cirrhosis can obtain improvement by using low dose of L-NAME.

Animals ; Liver ; parasitology ; Liver Cirrhosis, Experimental ; pathology ; physiopathology ; Male ; NG-Nitroarginine Methyl Ester ; therapeutic use ; Nitric Oxide ; physiology ; Rats ; Rats, Sprague-Dawley ; Testicular Diseases ; drug therapy ; etiology ; Testis ; drug effects ; pathology ; physiopathology ; Testosterone ; blood

OBJECTIVETo characterize the biological function of calmodulin (CaM) from Clonorchis sinensis (C. sinensis, Cs) and investigate its role in clonorchiasis-associated hepatic fibrosis.

METHODSThe full-length sequence of CsCaM gene was isolated from Cs cDNA library and its homologues were searched using BLASTx for comparison. Bioinformatics analysis was performed to compare the homologues and predict the physiochemical characteristics and functional domains. The gene was cloned in a prokaryotic plasmid and expressed in E. coli, and the recombinant protein was purified by affinity chromatography for immunizing rats to produce polyclonal antibodies, whose titer was determined using ELISA analysis. Immunoblotting analysis was carried out to determine of the purity and antibody recognition of CsCaM. Immunofluorescence assay was employed to analyze the tissue location of the protein. A rat model of liver fibrosis was established by introperitoneal injection of the recombinant protein.

RESULTSThe recombinant CsCaM protein obtained contained 150 amino acids with a theoretical molecular mass of 23.4 kD. CsCaM homologue had EF hand motifs. The recombinant pET-30a-CsCaM plasmid expressed in BL21 E. coli was about 23.4 kD. The total IgG antibody titer in the immunized mice reached the peak level (over 1: 51200) 2 to 4 weeks after the first injection. Immunohistochemistry showed that CsCaM located in the testis of adult C. sinensis. The rats receiving intraperitoneal injection of CsCaM showed severe liver inflammation with mild to moderate liver fibrosis.

CONCLUSIONThe pro-inflammation and pro-fibrosis effects of CsCaM in rat liver suggest its involvement in clonorchiasis- associated hepatic fibrosis.

Animals ; Antibodies, Helminth ; blood ; Antigens, Helminth ; immunology ; Calmodulin ; immunology ; Clonorchiasis ; immunology ; Clonorchis sinensis ; immunology ; Enzyme-Linked Immunosorbent Assay ; Gene Library ; Immunoglobulin G ; blood ; Inflammation ; Liver Cirrhosis ; parasitology ; Male ; Mice ; Rats ; Recombinant Proteins ; immunology