异种肝细胞脾内移植海藻酸钠包裹最佳浓度及免疫隔离机制的初步研究

Primary Study of Alginate Solution Consistence and Immunity Isolation Mechanism by using Transplanted Microencapsulated Xenogeneic Hepatocytes

目的:探讨微囊化异种肝细胞脾内移植对药物性肝衰大鼠的治疗过程中海藻酸钠包裹移植肝细胞的最佳浓度;观察受体存活率,测定肝功能及CD4,CD8的变化。方法:分别以浓度为5.0%、1.5%、0.15%的海藻酸钠悬液体外包裹经胶原酶技术制备的异种(豚鼠)肝细胞为供体,SD大鼠为受体,D-氨基半乳糖(19.5ml/kg)腹腔内一次性注射,制作肝衰模型。48h后将三组微囊化的游离豚鼠肝细胞(1.5×107个)分别移植入大鼠脾脏内。以生理盐水1.2ml脾脏注射为对照。移植后48h、96h观察存活率,肝功能、免疫球蛋白及肝脏病理情况,免疫组化ABC法测定CD4,CD8的变化。结果:微囊化肝细胞移植组存活率分别为50.0%、80.0%、90.0%,明显高于生理盐水组(25.0%)(P<0.01)。移植后96h测定总胆红素和AST显示:在不同浓度的三组微囊化移植肝细胞中,0.15%的海藻酸钠包裹组明显低于其它各组及对照组。移植后48h、96h,分别测定受体脾脏CD4,CD8淋巴细胞,0.15%微囊包裹组96h呈阳性,1.5%、5.0%包裹组48h均呈阳性。结论:大鼠药物性肝衰微囊化肝细胞脾内移植海藻酸钠包裹浓度为0.15%,细胞免疫参与排斥反应,包裹浓度过高影响移植肝细胞功能及免疫隔离效果。

Objective： To discuss the best consistence which is equal to encapsulated hepatocytes by alginate. We observed the change of CD4 , CD8, survival rate and hepatic function of xenogeneic encapsulated hepatocytes intraspleen transplantation （XHT） in the treatment of toxic hepatic failure （THF） in SD rat. Methods： Three groups rats received intraspleen graft of 1.5 × 10^7 guinea pig encapsulation hepatocytes by difference consistence such as 5.0% ,1.5% ,0.15% alginate packed at 48h after D - glactosamine （ D - GI） introperitonear injection（19.5ml/kg）, and N. S injection 1.2ml （N. S n =20 ） was belonged to control group. The results of CD4, CD8, survival rate and hepatic function, pathology were carried out at 48h,96h after transplantation. Results： There was a significant difference in survival rate at 96h between each XHT group （50.0% ,80.0% ,90.0% ） and N. S group （25.0% ） （p 〈0.01 ）. Up to 96h, In the 0.15% XHT group , total bilirubin and AST value was the lowest. There was significant difference among 0.15% XHF group and the other groups. CD4, CD8 lymphocytes in the recipient spleens by immunohistochemistry inspected at 48 h , 96 h past - graft respectively. In the 0.15% XHT group, it response CD4, CD8 lymphocytes at 96 h. In the 1.5% and 5% XHT group, There was masculine after 48h. Conclusion： The authors concluded that the best enwrap consistence of encapsulated xenogenic hepatocytes was 0.15%. Cellular immune response participates in the rejection. If the warp consistence is too higher , It would be a disbennifit to replant hepatic function and a diminution to immune response.