| PurposeIn our experiments,we researching for the macrophage's phagocytic ability,and T lymphocyte-mediated cellular immunity as well as associated cytokines produced,and B lymphocyte mediated the humoral immunity by the carbon clearance test,Immune imbalance models and hemolysin experiments.Finally,the use of ConA causing Immunological live injury model,in cellular and molecular level detecting the immune-related gene expression and discussing the protected mechanisms of ASPS on protecting the liver injury by mean of ELISA,RT-PCR,Western blot and other biotechnology.Methods1?Using of carbon clearance experiment to research the mononuclear macrophages function of immunosuppressive mice on ASPS,by calculating thymus and spleen indexes,K value and a value.2?Researching the activation of T lymphocytes and the ratio of CD4 +/CD8+ in peripheral blood lymphocytes on ASPS by using DTH model.Observing the HE staining in mice ear sliced as well as calculating thymus and spleen indexes,ear swelling and swelling rate and detecting the proportion of CD4 + and CD8 +inCD3 +by flow cytometry method.3.Researching on serum cytokine in DTH and immunocompromised model of ASPS.4?Researching the B lymphocyte proliferation on ASPS by calculating HC50 in hemolysin experiment.5?Using ConA to research the mechanism of immunological liver injury treatment with ASPS.Taking the liver tissue into pathology examination and pathology grade;Dectecting the content of AST,ALT,IL-2,IL-4and INF-? in serum and observing the content of NO,SOD,MDA,GSH-PX,IL-1? and TNF-a levels in liver homogenates;Measuring the expression levels of IL-2mRNA,IL-4mRNA and INF-?mRNA in splenic tissue and TNIF-amRNA,iNOSmRNA and ICAM-1mRNA in liver tissue by RT-PCR;Analysis the protein expression levels of TNF-a,ICAM-1,iNOS and NF-?B in liver tissue by Western blot.Results1?the thymus index,spleen index,K value and a value of model mice compared with the control group has a significant difference(p<0.01),The thymus index,spleen index,a value of ASPS high dose group and positive drug group compared with model group have increased significantly difference(p<0.01).ASPS median dose every index except thymus index compared with the model group were significantly increased(p<0.01),and other indicators like ASPS low dose had no significant difference(P>0.05).2?Model group Observed by microscope has lot of monocytes,eosinophils and neutrophils infiltration,tissue swelling greatly compare with control group,there is a significant difference.Positive drug DXM group and ASPS high dose group effectively inhibited ear swelling,and inflammatory cell infiltration,While every ASPS administration group make the ear tissue swelling degree more subsequently decreased,and a dose-dependent manner.Swelling of the model group with the control group,spleen and thymus index increased,with a very significant difference(p<0.01),the positive control group(DXM)and ASPS high dose group ear swelling in mice,the thymus index compared with the model group be greatly lower and have a significant difference(p<0.01).Others ASPS group swelling rate and the thymus,spleen index compared with model group be decreased,but no significant difference compared to control group(p>0.05).In DTH model group,the peripheral blood lymphocytes of CD4+lymphocytes relative levels increased,CD8+lymphocytes relative content decreased,CD4+/CD8+ratio was significantly increased,there was a significant difference compared with the control group(p<0.01),positive(DXM)group and ASPS high dose group compared with the model group,CD4+,CD8+proportion were reduced,CD4+/CD8+compared with model group has significantly decreased(p<0.01);ASPS in low-dose group and median dose group have no significant difference(p>0.05)compared to model group.3?In the DTH model,the model group increase IL-2,IL-4 and IFN-? content compared to control group have a very significant difference(p<0.01),positive drug DXM group and ASPS high dose group compared with the model group the content of cytokines IL-2,IL-4 and IFN-y has significantly decreased(p<0.05),ASPS median and low-dose group compared with the control group,the cytokine IL-2,IL-4,IFN-? content is also showing an decreasing trend has no difference(P>0.05).In immunocompromised model,The content IL-2,IL-4 and IFN-yof the model group was reduced comparing with control group,and has a highly significant difference(p<0.01),ASPS administration of each group compared with the model group,mouse serum cytokines IL-2,IL-4 and IFN-y correction increasing,especially the high-dose group ASPS comparison with the model group with a very significant difference(p<0.01),ASPS median dose group,ASPS low-dose group reveal the dose correlation.4?ASPS treatment group compared with the control group,every group of HC50 has tended to increase,and the ASPS high-dose group compared with the control group with a very significant difference(p<0.01),ASPS median and low dose group had no significant difference compared with the normal control group(p>0.05).6?ASPS can not only decrease lymphocyte infiltration,reduce swelling and necrosis of liver cells,but also increase the content of SOD,GSH as while as reduced the content of MDA,AST,ALT,IL-2,IL-4,INF-y,NO,IL-1?,TNF-a;ASPS group Can reduce the expression level of IL-2mRNA,IL-4mRNA and INF-ymRNA in splenic tissue,and reduce the content of TNF-amRNA,iNOSmRNA,ICAM-1mRNA and NF-?B in liver tissue,as well as reduce the ICAM-1,iNOS and NF-?B protein expression levels.Conclusion1?In this study,Analysing of the monosaccharide composition of GlcUA?rha?xyl?fru?gluc in ASPS by UPLC-MASS.Detecting the molar ratio of monosaccharides between the initial establishment of quality standards.2?ASPS has ability to activate the macrophages and enhance the body's non-specific immunity ability in immune suppression model,promote the expression and secretion of relevent cell factors;ASPS also can inhibit T cell proliferation and differentiation in DTH,as well as inhibit inflammation-related cytokine secretion and expression;In normal status,ASPS can activate the B cells and enhance the humoral immunity.Further explanation immunomodulatory effects of ASPS.3?The results show Acanthopanax polysaccharides ASPS has a protective functionon in immunological liver injury,which is not only reflected in directly decreasing aminotransferase protecting liver aspects,but also in removing free radicals,increase antioxidant enzyme activity,improve the body's anti-fat quality peroxidation.The mechanism may be associated with decreased secretion of inflammatory cytokines and adhesion and expression of related factors. |
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