The Transplantation Of Bone Marrow Mesenchymal Stromalcells For Treating Gout Kidney

Objective: The objective of this study was investigating the effects of the intravenoustr-ansplant ation of bone marrow mesenchymal stromal cells(BM-MSCs)on alleviati-ng epitheli al to mesenchymal transition(EMT)and promoting renal cell differenti-ation and gr owth and antioxidative stress in rats with gout kidney.Furthermore,t-he correspondi ng mechanisms were explored.Methods:1.36 immature male Wistar rats were divided into two groups randomly,one group named the control group had 12 rats,and the other group called the model group had 24 rats.The model group with gout kidney was established through a-de nineinducing for 4 weeks.2.After the gout kidney model was established successfully,the model group was randomly divided into PBS group and treatment group.Treatment group wou-l dbe injected with BM-MSCs,and the PBS group would be injected with phos-phat ebuffered saline(PBS).Then we got three groups,the control group,the PBS group,the treatment group.3.When made the gout kidney model,there were 6 immature male Wistar r-a ts which body weights were 50±10g.The rats were sacrificed for getting the bo-n emarrow,and the BM-MSCs were isolated,cultured and purified in vitro usingden sity gradient centrifugation combined with adherent screening method.We iden-tifi ed the cells by detecting their phenotype using the flow cytometer,and observetheir morphology,and induced their multiple differentiation potential for evaluatingB M-MSCs.4.After the models were established,we got the blood from eye sockets fordetec ting the serum creatinine(Scr)and blood urea nitrogen(BUN),at the same ti-me coll ected the rats urine to test 24 hours proteinuria.Then treatment groupratswere inje cted with BM-MSCs via tail vein,whereas the PBS group rats wereinj-ected with ph osphatebuffered saline(PBS).5.After six weeks for transplantation,urine and blood were detected again toas sess 24 hours proteinuria,serum creatinine(Scr)and blood urea nitrogen(BUN).Then killed the rats,we got kidneys to make the paraffinembedded specimens f-or HE,PA S and Masson staining.The staining renal tissues of three groups wereevaluated via using a semi-quantitative technique in a blinded fashion.6.Methods Immunohistochemistry was perfomed to observe transforming gro-wth factor-?1(TGF-?1).Western blot to determine protein expression of P-P38 /P38 and s elenium-containing enzyme thioredoxin reductase 1(Trx R1)in renal tissues.Results: 1.The rats of the model group which was induced by adenine,appeared the mass proteinuria,deterioration of renal function compared with the control group.And we observed big and white kidneys,the histopathologic injury in kidneys also appeared.Massive uric acid stones appeared at renal tubules and interstitium,renal tubular epithelial cells were damaged and became necrotic,Lymphocytes andmonocytes infiltrated,and formed the multinucleated giant cells and fibrosis in theinterstitium,that implied the gout kidney evolved as chronic renal failure(CRF).2.The third generation of cells were detected by the flow cytometer.CD90 was a positive marker which expression was higher than 95%,whereas CD45 wasused asa negative marker,its expression level was less than 4%.At the same ti-me osteogeni c differentiation was determined using alizarin red staining to exami-ne and got calcified nodules,whereas adipogenic differentiation was assessed usin-g oil red O staining and found lipid droplets.All proved that BM-MSCs were s-uccess fully isolateed.3.BM-MSCs improved renal function,the level of BUN and Cr and24 hoursprot einurine were reduced.Additionally,TGF-?1 elevation was cut down,at the t-ime,ac tivation of P38(i.e,P-P38 significantly decreased)in the treatment groupwas decreas ed obviously.Trx R1 were also increased compared withthe PBS group.Conclusion:BM-MSCs could alleviate the renal damages of adenine-induced gout kidney/chr onic renal failure,improve the renal function,promote renal cell differentiatio-n and growth and anti-oxidative stress by increasing the Trx R1 elevation,in addi-tion,dec rease the expression of TGF-?1 that contributed to alleviating EMT,whi-ch was me diated through supressing TNF-?/P38 signaling pathway.