Imaging And Immunohistochemical Research On Blood Supply Of Rabbit Vx2Liver Metastases

Objective: Bulit the model of rabbit liver metastases byartificially.Explain the changes of blood supply of liver metastases through thechange of CT perfusion,including hepatic arterial perfusion, portal veinperfusion,hepatic arterial index,portal venous perfusion index;the DSA ofdirect hepatic arteriography and indirect portography;immunohistochemicalanti-CD34antibody, anti-ICAM-1antibody, anti-α-SMA antibody changes.Materials and Methods: Forty New Zealand white rabbits,weight2.5~3.0kg,male or female. Exposed the spleen through the left upper quadrantabdominal incision,inject the the VX2tumor strains in the spleen. Confirm theliver is form metastases though CT scan after2weeks.The planting successfulrabbit perform CT perfusion,observe the changes of HAP,PVP,HPI and PPI,confirm the change of blood supply of liver metastases.The next day make theDSA examine,make direct hepatic arteriography and indirectportography,direct observation of the blood supply of liver metastases inarteriography and portography.Then anatomy rabbits,separation of rabbit livermetastases,dehydration and fixation, immunohistochemicalstaining.Anti-CD34antibody, evaluation of new blood vessels within thetumor; anti-cell adhesion molecule antibody (ICAM-1), evaluation of theexistence of the tumor around the sinusoidal; anti-smooth muscle actinantibody (α-SMA), evaluation of arterial neovascularization within thetumor.Observed the CD34, ICAM-1, α-SMA MVD at different stages ofchange, to evaluation the liver metastasis of CD34and ICAM-1, α-SMAchanges on the evolution of blood supply of liver metastases.Results:1.The establishment of40rabbit liver metastasis model,successful modeling33, anesthesia death2,unknown willing death3,unsuccessful planting2. 2.CT perfusion of liver metastases of HAF,HAP,HPI is greater than theperlesion liver tissue,PVP and PPI is less than he perlesion liver tissue.3.33planting tumor rabbits underwent DSA,found62liver metastases.Directhepatic arteriograph show hepatic artery directly supply perlesion,andappearance ring staining,zone of necrosis can not staining. There is5rabbitsperform indirect portography.It shows portal vein surround livermetastases,appearance ring defect,no signs blood supply of portal vein.4.Make131paraffin blocks of liver metastases immunohistochemistry. Theaverage size of metastases approximately315.76±24.78μm, liver metastasestumor expression of CD34and ICAM-1, while the positive expression the rateof ICAM-1more than the rate of CD34expression; the average size of themetastases of approximately632.58±35.46μ m, the CD34positiveexpression rate of metastases is increase, ICAM-1expression is reduced,α-SMA positive expression rate is discover in the tumor.;the average size ofabout2683.15±167.32μm,only CD34and α-SMA-positive expressionrate of intratumoral,ICAM-1staining disappeared,only few staining foundaround the sinusoidal tumor or normal sinusoidal.The average size more than1cm metastases,with the tumor grow,the center necrotic tissue becomemore,the CD34positive expression rate is still increase,ICAM-1stainingdisappeared,but α-SMA positive expression rate is not obviously increase.Conclusion:1.PlantingVX2in spleen form liver metastases correspondthe route of human portal vein metastasis,more conducive study the bloodsupply of hepatic metastases.2.CT perfusion scan show liver metastases HAF,HAP and HPI is greater thanperlesion liver tissue, PVP and PPI is less than perlesion liver tissue.Itindicate that the artery blood supply of liver metastases increase and the portalblood supply reduce.3.DSA results found that the liver metastases supply hepatic arterial blood ofliver metastasis.DSA direct hepatic arteriography shows the tumor vascularring staining, tumor center no or minor staining, indirect portal veinangiography shows the ring defect of tumor region, no signs blood supply of portal vein.4.Pathological immunohistochemical study found that the hepatic artery andportal vein blood supply of liver metastases in the early stages of livermetastases, with increase the size of liver metastases,the artery blood supplyof liver metastases gradually increased while the portal vein blood supplygradually reduce, liver metastases is only supply by artery blood when becomegreater than2683.15μm, portal vein does not blood supply.With the livermetastases growing, the center necrotic tissue become growing,there isvascularization and arterial neovascularization in tumor solid area.