A Experimental And Clinical Study Of Cyclophilin

Objective: Cyclosporin(CsA) has been used in immunologic diseases,which takes effect through Cyclophilin(CyP),the intracytoplasm receptor of CSA .In this article,we set up the new method to determine the CyP mRNA by RT-PCR firstly;secondly,we build the animal model of Adramycin Nephrosis (ADN),then we detected the CyP gene expression of nephridial tissue in ADN rats,we also test the total blood protein,albumn,cholestery,Urine Protein in 24 hours(24hUP), Blood urea nitrogen,creatinine,and the relationship between CyP mRNA and the blood and urine indexes mentioned as above.at the same time,we investigated the curative effect of CsA in different pathological types of nephrotic syndrome(NS), the clinical significance and the relation of CyP and the curative effect of CsA was studied.Methods: We found the most suitable method of RT-PCR to detect CyP by different reaction systems,through that method, we detected the CyP gene expression of nephridial tissue in ADN rats;83 cases patients that were diagnosed nephritic syndrome(NS) , orally administrated CsA 5mg/(kg·d) for 6 to 24 months with gradually decrement of glucocorticosteroid, meanwhile, the dose of CsA was adjusted according to CsA concentration in blood.The effect of CyP gene expression on the curative effect of CsA was studied.Results: By the RT-PCR method,the base pair of CyP was 408 bp. CyP andβ-Actin bands (234 bp) were seen distinctly in agarose gel electrophoresis without mixed strap.The CyP mRNA expression in nephridial tissue of ADN rats models was higher than the controls, and which was positive correlation with cholesterin and urine protein for 24 hours, and negative correlation with the blood total protein,albumin,there was no relation with renal function . The levels of CyP mRNA expression were found higher with mesangial proliferative glomerulo-nephritis(MsPGN) and focal segmental glomerulonephritis (FSGS) than in minimal change nephrotic syndrome (MCNS), there was no difference in the levels of CyP mRNA expression with steroid dependent, frequent relapsing and steroid resistant types. The levels of CyP mRNA expression in acute stage were also found to be higher in acute stage than in convalescent stage and in control groups. We get the better curative effect of nephrotic syndrome of different pathological types by CsA ,and The overall remission rate was 81.93 % .the renal toxicity appeared with high blood CsA concentration,when the concentration sustains at 100～200 ng/ml,and CyP level keeps stably, the NS will relieve.Conclusions: The method of RT-PCR for CyP we set up was a new and flawless means with high sensitivity and specificity ,and best reproducibility, and easy to perform, quickly get the result, which minimized the error and ensure the results accurate. The higher level CyP mRNA maybe one of the indexes of renal tissue damage. CsA is one of the effective substitutes for glucocorticoid for immunologic diseases,which could take effect quickly and effectively.this article provided the RT-PCR method and the theoretical evidence for clinical apply CsA and judgement of therapeutic effect and prognosis.