TMP Delay The Proceeding Of Chronic Allograft Nephropathy Of Rat Associating With Dan-ShenRoot

Objective:To explore the therapeutical effect of Tetramethylpyrazine and Dan-ShenRoot on Chronic allograft nephropathy of rat , reveal the relationship with CAN , then discovering mechanism of action that delaying the proceeding of CAN of rat,which provide a new way of therapy to extend survival time of transplanting kidney.Methods:1.We used Fisher rats as donors and Lewis rats as recipients to establish the model of CAN.The left kidney of the donors was removed,the inferiorvena of the donor was anastomosed end-to-end with the left renal vein of the recipient, and the donor's abdominal aorta was anastomosed end-to-side with that of the recipient with the unaided eyes to carry out renal allograft in prime position.2.125 rats with transplanting kidney were divided into five groups: Ciclosporin group, TMP associating with Ciclosporin group, Dan-ShenRoot ssociating with Ciclosporin group, TMP and Dan-ShenRoot ssociating with Ciclosporin group,black group; every group for 25 rats. Route of administration: Ciclosporin was intragastric administration, TMP and Dan-ShenRoot were intraperitoneal injection. Medication was used for 10 days after postop. During the postop periods, observed the life of rats, all rats were sacrificed at the 2nd,4th,6th,8th and 12th, examined the changes of HE,PSA,Masson staining was apllied to transplanting kidney of five groups. 3.Transplanting kidney were detected by real-time PCR and immunohistochemistry methods were apllied to detect TGF-β1 mRNA and protein expression in the transplanting kidney, revealed the mechanism of action that delaying the proceeding of CAN of rat.Results:1.The model of CAN of rat was establishedIt was marched 140 pou operation after technique techniqued, the ratio of operation was reached 96.43%.Cumulative time of operation was 150 min, recipe kidney was 20 min, transplantation was 70 min. The pathological changes of the transplanted kidney began to appear 4 weeks after transplantation ,then the chronic rejection could be seen after 12 weeks.2.Histopathological changes of Transplanting kidneyThe rats of black group was died in two weeks. Group A appearance the histopathological changes of CAN since 4th week, it is thus evident that typical histopathological changes of CAN at 12th week.But Group B,C occurrenced were late,Group D was later. There was significant difference between group A and others (p<0.05); between group A and D has predominance difference(P<0.01);there was no significant difference between group B and group C (p>0.05).3.TGF-β1 mRNA and protein expression in transplanting kidney detected by real-time PCR and immunohistochemistry4 weeks after transplantion, masculine cells with TGF-β1 began to increase,which of group A increased obviously,group B and C were lower,group D lowest.The expression of TGF-β1 enhanced by the time of transplantation.At 12 week,A group enhanced obviously,D group was lowest.Then we made statistics analyse during every group,group A compared with other groups it had statistical significance(P<0.05),the difference between group A and D was more significant(P<0.01); It had no statistical significance that group B compared with group C (P>0.05).Through the expreesiom of TGF-β1 detected by real-time PCR in transplanted kidney of rats after transplantion,we found it group A increased obviously than other groups(P<0.05), the difference between group A and D was more significant(P<0.01); It had no statistical significance that group B compared with group C (P>0.05).Conclusion:1. This method was simple and direct-viewing,and reduced microsurgery equipment,which successfully established rat renal transplantation model of chronic allograft nephropathy and can achieve the requirement of establishing CAN models.2. The together use of TMP and Dan-ShenRoot was safe,reliable and had lower side effect,then slowed the proceeding of CAN.3. TMP and Dan-ShenRoot used together can decline the expression of TGF-β1 ,then it could play effect of anti-fibrous degeneration of CAN,and slow the proceeding of CAN.