| The adhesion of leukocyte to vascular endothelial cell is the key step of leukocyte migration and lymphocyte recirculation, and plays a essential role in inflammation and immune response.HuangLianJieDuTang (HLJDT) and it's major chemical component berberine,baicalin,gardenoside have the function of anti-inflammation and immune regulation and others.Therefore,the function of HLJDT is may be related to the synergy effects of it's major chemical components berberine,baicalin,gardenoside.Based on the study results of the HLJDT and it's major chemical components,the prescription of QingJingNing (QJN) was consist of the major components of HLJDT in this experiment.The influence of QJN on the adhesion of leukocyte to endothelial cell and adhesion molecules expression was investigated in vivo and in vitro,so as to research the machanism of anti-inflammation and immune regulation function of QJN .1 Experiments in vivo1.1 The effects of QJN on immune function in SRBC-immunized miceTo study the effects of QJN on immune function in mice with high dose(14mg/kg),middle dose (7mg/kg) and low dose (3.5mg/kg) . The animal model was made by immuning mice with sheep red blood cell (SRBC),delayed type hypereneitivity (DTH) by measuring foot thickness,quantity of plaque forming cell (PFC) by hemolytic spectrophotometry, proliferation of T and B lymphocyte by MTT colorimetry, phagocytosis of Mφ in abdominal cavity by neural red colorimetry were observed respectively. The results demonstrated:(1)Three dose could significantly decrease DTH index in mice (P<0.05).(2)High dose and middle dose both could siginificantly inhibit PFC(P<0.05,P<0.01), low dose had no effect(P>0.05). (3) Three dose could significantly inhibit proliferation of T lymphocyte induced by ConA (P<0.01).(4) Three dose could significantly inhibit proliferation of B lymphocyte induced by LPS (P<0.01).(5) Three dose could significantly decrease the phagocytosis of Mφin mice.The results indicates that QJN has siginificant effects of anti-inflammation and immunosuppression.1.2 The effects of QJN on cytokine in SRBC-immunized miceTo investigate the effects of QJN on IL-1,IFN-γ,IL-2 in SRBC-immunized mice. The IFN-γlevel was measured by macrophage NO2ˉ-release assay, the IL-1 level was measured by thymocyte assay, the IL-2 level was measured by mitogen activated lymphocytoblast assay respectively. The results showed:(1)Three dose could significantly decrease IL-1 in serum (P<0.05,P<0.01),and also could inhibit macrophage to secrete IL-1 induced by LPS (P<0.05,P<0.01).(2) Three dose could significantly decrease IL-2 in serum (P<0.05,P<0.01),and also could inhibit splenocyte to secrete IL-2 induced by ConA (P<0.05,P<0.01).(3) Three dose could significantly decrease IFN-γ inserum (P<0.05,P<0.01),and also could inhibit splenocyte to secrete IFN-γ induced by ConA (P<0.05,P<0.01).The results suggests that the inhibition effects of QJN on immunocyte to secret IL-1,IFN-γ and IL-2 could be associated with anti-inflammation and immunosuppression.2 Experiments in vitro2.1 The effects of QJN on the adhesion of leukocyte to HUVEC The highest concentration of QJN without effect on cell activity was 117μg/ml determined by the effect of QJN on activity of human umbilical vein endothelial cell (HUVEC)and human peripheral blood mononuclear cell (PBMC) by MTT assay,and the effect of QJN on activity of human peripheral blood polymorphonuclear(PMN) cell by trypan-blue stain assay.Based on this result,the effects of QJN with high (117μg/ml),middle (58.5μg/ml),low(29.25μg/ml)concentration on the adhesion of PMN,PBMC to HUVEC TNF-induced were oberserved. The results showed:(1)After 2h and 12h HUVEC treated with high and middle concentration QJN,both could siginificantly decrease the PMN-HUVEC adhesion (P<0.05,P<0.01), after 2h HUVEC treated with low concentration QJN ,QJN could siginificantly decrease the adhesion of PMN to HUVEC (P<0.01),but had no effect after 12h (P>0.05).(2) After 2h and 12h HUVEC treated with high c...
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