| Objectives:To investigate the effects of the extracts from Phyllanthus urinaria L. on prevention of Schistosoma japonicum ova-induced granulomas and liver fibrosis, and to clarify its possible molecular mechanism.Methods:21 healthy and adult male Balb/c mice were randomly divided into 3 groups, Treated group (infected/the extracts from Phyllanthus urinaria L.-treated mice), Model group (infected/untreated mice), and Normal group(uninfected mice),7 mice per group. Treated group and Model group were infected by cercariae of schistosoma through the skin on the same day, via 6 weeks to form chronic infection model. After six weeks treated group and model group uesd the praziquantel to kill the insect. In Treated group, the extracts from Phyllanthus urinaria L. (100mg/kg·d) were administrated after insect disinfestation,and physiological saline was administrated the other groups (30 days of successive administration). After administration all mice were sacrificed. A portion of liver tissue were fixed in 4% paraformaldehyde in PBS, and another the other portion of the liver tissue were frozen in liquid nitrogen tank. Remove the spleen, isolate the spleen cells for RNA extraction. HE staining was used to assay pathological changes in liver,and to compare the size of egg granuloma. Masson trichrome staining for collagen was used to evaluate the score of hepatic fibrosis in each liver tissue section. Alkaline hydrolysis for measuring Hyp in liver tissue and ELISA for measuring IL-13 in homogenate of liver tissue. RT-PCR for measuring the expression of GATA3 mRNA in spleen cells. Quantitative real-time RT-PCR (q-RT-PCR) was used to quantitate the mRNA expression of Argl in liver tissue.Results:1. Under the light microscope, Liver sections from the normal mice with HE staining shows that the structure of hepatic lobules was obviously complete,the liver cells arranged rule, and there were not egg granuloma, not cell denaturation, not inflammatory cell infiltration. In the model group, the liver cells were severe damaged and showed vacuolar change. Enormous egg granulomas surrounded by an inflammatory cell accumulation and fibril aggregation. In contrast, the Treated group with smaller granulomas,less liver cells denaturation and less inflammatory cells infiltration; The connective tissues were decreased obviously. Masson trichrome staining revealed liver sections from the normal mice showed no fibrosis. The liver sections from the model mice showed the collagen distributed around granulomas, liver fibrosis increases in the portal tracts.And part of them the liver fibrosis forms interlobular tissue. But compared with the model group the pathological changes of Treated group were reduced,with less significant fibrosis. The collagen around granulomas and limited to them.2.The production of Hyp in liver:Increased Hyp levels were measured in the model group in comparison to the normal group (P<0.01). And, the level of hepatic Hyp in the Treated group was significantly reduced(P<0.05)compared with the model group.3.The level of IL-13 in liver:in the model group,the levels of IL-13 were higher than that of the normal group(P<0.01). In the Treated group, the levels of IL-13 were lower than that of the model group (P<0.05).4. Detection of GATA3 mRNA:The expression of GATA3 in the model group was significantly enhanced than the normal group. The expression of GATA3 in the Treated group was significantly reduced than the model group.5. Detection of Argl mRNA:The expression of Argl in the model group was significantly enhanced than the normal group(P<0.01), The expression of Argl in the Treated group was significantly reduced than the model group (P<0.05).Conclusions:The extracts from Phyllanthus urinaria L. might decrease the level of Th2 associated profibrotic cytokines IL-13, and to down regulate the transcription of GATA3 mRNA in spleen cells and Argl mRNA in liver tissue. They alleviate the hepatic fibrosis caused by egg granuloma in Schistosoma japonicum infection... |
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