Treatment Research Of LESWT And BMSCs On Chronic Cyclosporine Nephropathy In Rats

Background:Shock wave is a mechanical wave,which has been put into clinical practice as a physical therapy method for more than 30 years,developing from the initial extracorporeal lithotripsy therapy,to bone nonunion and pain therapy.Especially the improved therapeutic effect on chronic ischemic diseases by low-energy shock wave found in the last century made the application range of shock wave wider and wider.Chronic kidney disease(CKD)is a frequently-occurring disease in our country,with about 120 million patients with chronic kidney disease in it.But at the same time,most of renal diseases,especially chronic renal insufficiency,are still lack of effective therapeutic means.Renal microcirculation disorders and tubular interstitial fibrosis play important roles in the development of CKD.There is no report on effective therapy for chronic renal insufficiency by the low-energy shock wave,as a new treatment means.Stem cell therapy is a hotspot of medical research in recent years.Recently,there are a lot of reports about basic and clinical research on stem cell therapy for CKD.Stem cell transplantation has been proved to be of certain curative effect in acute kidney injury,renal transplant,renal interstitial fibrosis and ischemic kidney diseases.Shock wave,in vitro,has been reported to strengthen differentiation capacity of stem cells,but no relative literature shows that it can strengthen effect of stem cell therapy in vivo.In order to confirm the above hypothesis,we select chronic cyclosporine nephropathy model as treatment object.Cyclosporine,a common drug in the field of kidney disease,is mainly used for treatment of kidney transplant rejection,refractory nephrotic syndrome and other diseases,but its side effects are that it is likely to cause chronic cyclosporine nephropathy,with the main pathological features of focal and stripped renal interstitial fibrosis,afferent vessel hyaline degeneration,inflammatory cell infiltration and renal tubular atrophy.Besides,ischemic injury is one of the main mechanisms of renal toxicity.These characteristics are advantages of low-energy shock wave and mesenchymal stem cells therapy.Therefore,we select,in this study,cyclosporine nephropathy rat model,with the intervention of low-energy shock wave and mesenchymal stem cells,to observe therapeutic effect on CKD by low-energy shock wave and stem cell technology,and meanwhile observe synergistic effect between low-energy shock wave and stem cell technology,and explore the feasibility of low-energy shock wave applied to the therapy of CKD and its prospect applied with stem cells.Objectives:To observe therapeutic effect on chronic cyclosporine nephropathy by low-energy shock wave and stem cell technology and the synergistic effect between them and to initially discuss its mechanism.Methods:1.Use shock waves with different energy levels(6 KV,8 KV,10 KV and 12KV)for intervention of normal SD rats and dynamically monitor their renal function(serum creatinine and urea nitrogen)and rat growth change.Cut kidney on 28 d for tectology evaluation.2.Divide experimental animals into 5 groups: Group C: normal group(control);Group H: chronic cyclosporine nephropathy group;Group L1: 100 intervention by 8KV shock wave;Group L2: 200 intervention by 8KV shock wave;and Group L5: 500 intervention by 8KV shock wave.The low-energy shock wave therapy groups with different intervention times are to dynamically monitor renal function(serum creatinine and urea nitrogen)and rat growth change.Cut kidney on 28 d for tectology evaluation,meanwhile,measure the expression level of some cytokines.3.Divide experimental animals into 5 groups: Group C: normal group(control);Group H: chronic cyclosporine nephropathy group;group L: group of low-energy shock wave intervention;Group B: mesenchymal stem cells therapy group;Group BL: group of low-energy shock wave and stem cell therapy.Dynamically monitor renal function(serum creatinine and urea nitrogen)and rat growth change,take kidney tissue for tectology evaluation on 28 d,meanwhile,test the expression levels of some cytokines.Results:1.Compared with control group,rats in low-energy shock wave therapy groups(groups L6 and L8)have no significant changes in the kidney tectology,rats in group L10 group have few visible renal extravasated blood on kidney tissues,rats in group L12 have some visible ecchymosis on skin,with some visible hemorrhagic spots scattered on the surface of the kidney,many extravasated blood in kidney tissues and cloudy swelling in renal tubules.Compared with normal control group on renal function and body weight,difference of serum creatinine,urea nitrogen and body weight change trends between groups of different energy-level shock waves is not significant.For renal interstitial fibrosis(RIF)score,compared with control group,RIF scores of low-energy shock wave therapy groups(groups L6 and L8)have no obvious difference,while RIF score in high-energy shock wave groups(groups L10 and L12)are higher than that of normal group,with statistically significant difference.There is no significant change for expression level of cytokines ?-SMA,TGF-?1,NF-?B and VEGF between the normal control group and low-energy shock wave therapy groups(groups L6 and L8),while expression levels of those cytokines in high energy shock wave groups(groups L10 and L12)are increased.2.Compared with group C,body weight,serum creatinine and urea nitrogen for rats in group H and shock wave therapy groups(groups L1,L2 and L5)are significantly increased,with statistically significant difference.Compared with group H,for rats in groups L1 and L2,body weight and the renal function are improved,serum creatinine and urea nitrogen are significantly dropped and body weight is increased.But renal function and weight of rats in group L5 are not improved,while serum creatinine,urea nitrogen and body weight have no significant difference compared with those of model group rats.For kidney tectology,rats in model group and therapy group show significant kidney damages and different degrees of tubular interstitial fibrosis,with RIF scores significantly increased.Compared with group H,rats in groups L1 and L2 have significantly alleviated interstitial fibrosis,with RIF score dropped significantly.But levels of RIF score is significant increased in group L5.For mechanism exploration,expression levels of cytokines closely related to kidney disease development,i.e.?-SMA,TGF-?1,NF-?B and VEGF,are significantly increased in model group and intervention group.Compared with group H,expression levels of the above cytokines in groups L1 and L2 are dropped,while expression levels of the above cytokines in groups L5 are increased.3.Compared with group C,group H and therapy groups(groups L,B and BL)have serum creatinine and urea nitrogen significantly raised and body weight significantly dropped,with statistically significant difference.Compared with group H,rats in groups L,B and BL groups are improved on body weight and renal function,with serum creatinine and urea nitrogen levels significantly dropped and body weight gained.For kidney tectology,both model group and therapy group show significant kidney damage and different degrees of tubular interstitial fibrosis,with RIF scores significantly increased;and compared with group H,rats in groups L,B and BL have interstitial fibrosis significantly alleviated,with RIF scores dropped significantly.For cytokine expression,?-SMA,TGF-?1,NF-?B and VEGF in model group and intervention group are significantly increased.But improvement in kidney function,body weight growth,RIF and cytokine expression level in the groups BL and B have no significant difference.Conclusions:Moderate low-energy shock wave and mesenchymal stem cell therapy can improve health of rats with chronic cyclosporine nephropathy.Synergistic effect of both is not found in the body,and the mechanism of kidney function improvement by both still needs further research.