| Objective:To develop a rat model of heterotopic heart transplantation with high postoperative survival rate by using the simplified surgical technique, for further studies on transplantation immunology, pathology and organ protection. Using the heterotopic abdominal cardiac transplantation rat model to investigate the anti transplant rejection function of PTD-△hNFATminiDBD fusion protein in vivo.Methods:Development of abdominal heart transplantation took place on the base of Ono's procedure. The operation on recipients went first so that the vessels could be anastomosed right after donor's heart cut off. Donor's temperature decreased by saline ice cube placed in abdomen until its axillary fossa felt cold, then the thoracic cavity was opened to stop heartbeat quickly by saline ice water. Whole process was mainly completed by a single man without microscope assistance. Male Wistar rats served as donors and male SD rats as recipients, the models of heterotopic abdominal heart transplantation in rats were copied successfully. Two strains that contain PTD-△hNFATminiDBD and△hNFATminiDBD plamid seperately were propagated, then induced to express the fusion proteins for further step of purification. Thirty-six models were divided randomly into four groups:normal saline treated group,△hNFATminiDBD treated group (control), PTD-△hNFATminiDBD treated group (experimental group) and cyclosporin (CsA) treated group (positive control). All recipients with successful operation were given routine stimula, meanwhile, normal saline, PTD-△hNFATminiDBD,△hNFATminiDBD, and CsA were given into the rats everyday for 7 days by intraperitoneal injection respectively. The survival time of donor's heart in four groups was recorded for further statistical analysis, except one of each group was selected for pathological section at seventh day post-transplantation.Results:The model of heterotopic abdominal heart transplantation in rats was copied successfully by the modified Ono method. The success rate of abdominal heart transplantation was up to 90.2%(37/41). The mean value of survival time of donor's heart of no treatment group was (8.63±0.95) days,△hNFATminiDBD group was (9.06±1.32) days, PTD-△hNFATminiDBD group was (15.06±0.94) days, and CsA group was (22.94±1.15) days. The mean value of survival time of donor's heart was significantly prolonged by PTD-△hNFATminiDBD fusion protein but it's still shorter than that of the CsA group. (ANOVA, P<0.05)Conclusion:The advantage of Developed abdominal heart transplantation lies in simplifying the procedure and succeeding highly. The method is convenient specifically for those researchers whom haven't experience on vascular anastomosis. PTD-△hNFATminiDBD fusion protein can prolong the survival time of donor's heart in rats. It may open up the additional avenues for novel therapeutics for transplant reject reaction and autoallergic diseases. |
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