The Effects Of Ligation Intervention Blood And Lymphatic Vessels Of The Rat Walker-256Cell-burdened Lymph Nodes

Objective:（1）Histologically characterize angiogenesis and lymphangiogenesis in metastasticregional LNs proximal to tumors.（2）We observe the Morphology and distribution traits of angiogenesis MVD andlymphangiogenesis MLVD in the rat popliteal fossa of Walker256cell metastaticlymph nodes after the ligation intervention of lymph circulation and bloodcirculation,to explore its possible role in progression and metastasis.（3）Further discuss the mechanism of skip metastasis in malignant lymph nodes.Methods:1.70SD rats were from the Department of Animal Laboratory in South Chinaof University. To establish xenograft and popliteal lymph node metastasis model,Walker-256cells were injected in the bilateral plantar of70SD rats.6SD rats wereselected to transplantation experiment,64SD rats were selected and randomlydivided into4groups: ligation of lymph ducts as group1, ligation of vasculars asgroup2, ligation of lymph ducts and vasculars as group3，the parental group as group4. We Measured all the popliteal lymph nodes’ size before the ligationintervention,statistical analysis showed no significant differences2.After surgery Day3, Day7, Day14and Day21,1）we observe the size,volume and weight of the rat metastastic popliteal fossa lymph nodes;2）By HEstaining, observe the structural changes in the rat popliteal fossa lymph nodes,detection rate of malignant embolus;3)Immunohistochemistry: D2-40and CD34double labeling immunohistochemical staining was used in4groups to scan anddetect the changes of lymphangiogenesis MLVD and angiogenesis MVD.Results:（1）Compared with the control group, after ligation intervention, every time points of lymph nodes’ volume were reduced（P＜0.05）；Comparison between the groups,every time points of lymph nodes’ volume of Group3(ligations of all the input andoutput lymph ducts and vasculars)was significantly reduced (P <0.05).（2）The success rate of Walker-256rat sarcoma metastasis popliteal lymph nodesmodel was96.09%, confirming the model’s success rate was high.（3）HE pathological observation:Structural damage in each group, transplantmetastasis lymph nodes were atrophy obviously and partial necrosis. Tumor thrombuswere found in the intravascular or lymph vessel of metastatic lymph nodes, and7inGroup1（21.87%）,5in Group2（15.62%）,2in Group3（6.26%）,17in Group4, analysis between the groups was statistically significant P <0.05.（4）After rat Walker-256breast cancer sarcoma popliteal fossa metastasis lymphnodes were ligated,3day,7day,14day,21day time point of CD31and7day,14day,21day of D2-40double labeling immunohistochemical staining showedthat MVD and MLVD values of Group1,2and3were lower than the control group4(P <0.05). Comparison in groups, MVD and MLVD of each group were significantlyreduced with time passed (P <0.05), the MVD of Group2reduced more than Group1(P <0.05). And the MLVD of Group1reduced more than Group2(P <0.05).Conclusion:（1）SD rats popliteal fossa metastasis lymph node of Walker–256breast cancerwere a ideal model.（2）Ligations of lymph ducts and vasculars reduce the lymphangiogenesis andangiogenesis in metastasis lymph nodes.（3）Ligations of lymph ducts and vasculars reduce the rate of malignantembolus.