The Intervention Of BMSCs To Adriamycin Nephrosis Rats And MTOR Expression

Object: To separate and cultivate BMSCs; and defining the relatedbiological characteristics of BMSCs.To build model of adriamycinnephrosis rats by injecting ADR.To compare the effects of cuingadriamycin nephrosis rats by transplanting BMSCs through biochemicalDetection, electron microscopy observation and observation by PCR andWestern blot. And researches in the intervention of BMSCs to adriamycinnephrosis rats and mTOR expression.Methods: To extract, cultivate and identify the SD BMSCs: to separate asingle nuclear cell layer from Bandicoot percoll, then cultivate themsingly by density gradient centrifugation and attachment culture. Finally,to identify the facial materials by flow cytometer and identify BMSCs byossification and adipogenesis induction.To build model of adriamycinnephrosis rats: to inject ADR7.5mg/kg under non-narcotic situation and14days later, composite all kinds of results:general behaviors, detectionof biochemical indicators such as24h urine protein quantitative andcreatinine and justification of the model by observing HE coloration ofkidney tissue and by Electron microscopy morphology.The interventionof BMSCs to adriamycin nephrosis rats and mTOR expression:experiment groups: model group, MSC group and normal group, todynamically observe the mRNA level and protein expression of slit diaphragm nephrin, nephrin of mammalian target of rapamycin and othernephrins at different time.Results: The rat BMSCs cells cultivated by density gradientcentrifugation and attachment culture can be easily divided and cultivated.They have surface antigen of leydig cell, that is high-purity, high-activity,multi-lineage differentiation. They are Completely accords with thebiological characteristics of rat BMSCs.14days later, after analyzing allkinds of results, the model made by this method is successful, and it canbe experimented. Comparing model group with MSC group, foot processfusion and microvilli degeneration of sertoli cell in kidney tissue aredecreasing obviously. MSC plays a protective role in cellular-structuredamage caused by adriamycin.,Comparing model group with MSCgroup, the expression of nephrinm RNA in kidney cortex is increasingobviously, the expression of mTOR is decreasing obviously,Comparingmodel group with MSC group, the2w and Western blot showed that theexpression of nephrinin creasing obviously,the expression of mTOR andits related proteins are decreasing obviouslyConclusions: Rat BMSCs of high-purity, high-activity, multi-lineagedifferentiation can be seperated from bone marrow of rats by densitygradient centrifugation and attachment culture. And they are accord withbiological characteristics of Rat BMSCs.The model can be made byinjecting ADR7.5mg/kg into caudal vein disposable.14days later, the model shaped.According to the results of this experiment, the mechanismof MSC protecting adriamycin nephrotic rats is that the injection of MSCsuppressed the activation signal path of mTOR in kidney texture whichkeeping the normal expression of nephrin. The normal expression ofnephrin maintains the completion of membrane structure of Sertoli celland they improve proteinuria and protect glomerulus from the damage ofADR. All these achieve the purpose of slowing down the diseaseprogression.