The Research Of The Expression Of Perforin And Granzyme B MRNA In Peripheral Blood During The Acute Rejection

Investigate the expression of perforin and granzyme B mRNA in peripheral blood during the acute rejection. First, improve technique for establishing a stable and easy replication model of heart transplantation in rat for the further research of acute rejection. Second, measure the expression of perforin and granzynie B mRNA in peripheral blood dynamically after heart transplantation in rat, in comparison with the pathological examination, in order to judge the relationship between the expression level and the acute rejection. Third, observe the expression level of perforin mRNA and granzyme B mRNA in the peripheral blood during the acute rejection after renal transplantation. Judge the worthiness of making the diagnosis of acute rejection and the value of evaluating the effective of anti-rejection therapy.MethodI. The establishment of heart transplantation in ratTest Group (n=16) the donor is the Wister and the SD as recipient respectively, the Contrast Group(n=17): SD was used as donor and recipient. Exposed the recipient's abdominal artery at first, obtained the donor's heart and up-right lung while the organs were in flowed, and the donor's aortic ascent artery was anastomosed to recipient's abdominal artery. The successful standard is the survival time above 24 hours.II. The expression of perforin and granzyme B mRNA in peripheral blood in acute cardiac allograft rejection in ratsTook the peripheral blood at variety time, at the meantime harvested the graft to observe the pathological changes. The blood samples were checked by the method of fluorescent quantitative reverse transcription polymerase chain reaction to measure the level of perforin and granzyme B mRNA.III. The clinical observation32 cases were investigated, who had suffered acute rejection after renal transplantation. We took the peripheral blood just after the transplant operation, and at variety time during the period of anti-rejection therapy. We check the serum creatinine and still used the method of fluorescent quantitative reverse transcription polymerase chain reaction to measure the level of perforin and granzyme B mRNA.ResultsI. The establishment of heart transplantation in rat80 heterotypic heart transplantations were performed, 4 died of anastomotic bleeding, 2 died of the stenosis of anastomsis and 1 died of the failure of heart rebound, and the total successful rate was above 80%. There is no difference between the test group and the contrast group (p > 0.05 ) .II. The expression of perforin and granzyme B mRNA in peripheral blood in acute cardiac allograft rejection in ratsAt 24 hours after transplantation, the expression of perforin and granzyme B mRNA began to up-regulate, reached the climax at 3 to 5 days, and still kept the high level during 7 to 11 days. They were significant difference to the level of pre-operation and the expression level of contrast group (p < 0.001). The test group's pathological examination showed I degree at 3-5 days, and ff-IV degree at 7-11 days. No changes could be observed in contrast group (p>0.05) .m. The clinical observationThe level of perforin and granzyme B mRNA was significant higher when the patient suffered acute rejection (p<0.001), and dropped down after effective anti-rejection therapy, the level of serum creatinine recovered also, but late to the drop of perforin and granzyme B mRNA.ConclusionI. Modified surgical technique and careful operative managements can contribute to success in heart transplantation in rats.II . The graft rejection process companied with the up-regulated expression of perforin and granzyme B mRNA in rats. The measure of perforin and granzyme B mRNA is maybe a good indicator of acute rejection.HI. Using fluorescent quantitative reverse transcription polymerase chain reaction to measure the expression of perforin and granzyme B mRNA in the peripheral blood offers a noninvasive means of diagnosing acute rejection of renal allograft, and it is useful to judge the effective of anti-rejection therapy.