Inhibition Of TF Expression In Islet Grafts In Vivo IBMIR Models Of Acute Research

Objective:In clinical islet transplantation, the instant blood mediated inflammatory reaction (IBMIR) is a major factor contributing to the poor initial engraftment of the islets. This reaction is triggered by tissue factor (TF) expressed by the transplanted pancreatic islets。In this experiment, Antisense RNA was constructed to inhibit expression of TF in porcine islet cell, use intraportal pig islet xenotransplantation into athymic mice as an in vivo model for he study of the instant blood-mediated inflammatory reaction, then The Effect of inhibition of Expressing TF in islet cell was observed for IBMIR after transplantation.Methods:Antisense RNA was designed to transfect porcine islet cell with antisense technology, then the realtime-quantitative reverse transcription-PCR were applied to select the best Antisense RNA for TF silencing; the indexes(the levels of TAT^ blood glucose and C peptide)were detected in vivo model of IBMIR in the porcine islet cell after TF silencing, to known the level of development of IBMIR. Hepatic samples were collected for Hematoxylin-Eosin staining and immunohistochemistry staining analysis.Result:The oligoTF-1006 which had the best effect of silence was selected by PCR detection; the realtime- quantitative PCR showed that the results of TF 1006-mRNA seliencing after 4 days by transfection of Antisense RNA in islet cells was the best; porcine islets after TF silencing were implanted into the portal vein of SCID mice, then the indexes (the Decine of blood glucose level in one hour,the diversification of C peptide and TAT) showed that the development of IBMIR in this group were lighter than those in control group and in blank transfection group. Conclusion:Tissue factor is the main initiation factor IBMIR. Antisense RNA can inhibit the expression of TF, TF expression in the silence of porcine islets transplanted via the portal vein of SCID mice can reduce the early activation of coagulation and platelet activation and reduce the development of IBMIR after replantaion of islet cells. This method can be used to lower the lost of grafts and increase the survival rate of islet cells clinically in the future.