Experimental Studies Of Inimicus Japonicus Venoms On The Immunomodulation And Erythrocyte Of Rat

Objective: The injury stung by Inimicus japonicus is an professional disease for fishermen, which cause intense pain, swelling, severe damage and hemorrhage around the wounded tissues, accompanying some immune systemic symptoms. In order to identify the mechanism of the damage, we investigated the cellular immunity and nonspecific immunity, observed the effects of the venoms on the immune function and structure of the macrophagocytes of the abdominal cavities in vitro. The effects of the venoms on thymus and spleen of rats were also investigated in vivo. We studied the effect of the venoms from Inimicus japonicus on fluidity of erythrocytemembrane lipids and structure of rat erythrocyte in vitro, and further investigated the preventive function of VitC (inhibitor of reactive oxygen species) on the damage. The experiments aimed to find the methods of treating the injury.Methods: We observed the effects of the venoms from inimicus japonicus on the immune function by determining the phagocytic function and structure of the rat macrophagocytes in vitro and in vivo. The phagocytosis of rat macrophages was evaluated by neatral red method and chicken erythrocytes method. The proliferative ability of macrophagocyte was determined by MTT method and SDH cytochemical staining. The effect of the venoms on the SP expression of macrophages and the immune organs of rats were detected using immunocytochemistry staining. The morphological changes of macrophagocytes and erythrocytes were observed by electricitive microscopy and HE staining. The damaging effects of the venoms were determined by fluidity of erythrocytemembrane lipids method. The content of MDA and the activities of GSH-Px, SOD, and T-AOC in the skin were examined by biochemistry and immunohistochemistry.Results:The results in vitro were as following: ( 1 ) The venoms could decrease the phagocytic function of macrophagocytes. Compared with control groups and VitC treatgroups, the venom groups phagocytic function decreased (P<0.01; P<0.01). Compared with the venom groups, VitC treat groups phagocytic function increased (P<0.01). (2 ) SDH cytochemical staining and the MTT activity of macrophagocytes: Compared with control groups and VitC treat groups, that of venoms groups decreased (P<0.01). Compared with venom groups, that of VitC treat groups increased (P<0.01). ( 3 ) Compared with control groups, the SP expression in vitro of macrophagocytes in venom groups decreased (P<0.01; P<0.01). Compared with venom groups, the differences of VitC treat groups were not detected. ( 4 ) The morphological changes of the macrophagocytes were observed by electricitive microscopy . VitC (inhibitor of reactive oxygen species) had preventive function to the damage in varying degrees. (5) The different doses all caused spina or swelling change of erythrocyte in varying degrees after 10min, 30min. 60min and 90min of the venoms administration, the damage of erythrocytemembrane were significant. Compared with the control group, the differences were significant in various venom groups (P<0.01). Compared with control groups and VitC treat groups , the fluidity of erythrocytemembrane lipids of venom groups decreased(P<0.01). Compared with venom groups, that of VitC treat groups increased(P<0.01).The results in vivo were showed as following: (1) Compared with control groups and VitC treat groups, the phagocytic function of venom groups decreased (P<0.01;P<0.01). Compared with venom groups, that of VitC treat groups increased (P<0.01). ( 2) Compared with control groups and VitC treat groups, staining and the activity of macrophagocytes of venom groups decreased (P<0.01). Compared with venom groups, VitC treat groups increased (P<0.01). (3 ) Compared with control groups, the SP expression of macrophagocytes of venom groups in vivo increased (P<0.01; P<0.01). Compared with venom groups, the differences of VitC treat groups are not significant. ( 4) Compared with control groups, the SP expression of the thymus and spleen in vivo of venom groups increased (P<0.01). Compared with venom groups,...