| Objective:To investigate effect of embryonic kidney mesenchymal stem cells on the expression of Nephrin and Podocin in the model rats of adria-Mycin-induced nephropathy(minimal-change type).Methods:The rats were divided into three groups randomly:model group,control group and RIMM-18 group.Model group and RIMM-18 group were injected adriamycin of one time via caudal vein.At the same time,control group were injected nomal Saline.After 3 days we gave rats of contral group 1ml culture medium via caudal vein and rats of RIMM-18 group were injected 1×10~7 RIMM-18 cells.At 6d,17d and 31d after the model being manufactured,the urine and blood of rats were collected to examine urine protein in 24h,serum albumin and serum cholesterol.The kidney of all rats were collected to examine pathological change of renal tissue and observe the expression of Nephrin and Podocin which were marker proteins of podocyte through technique of immunofluorescence and Western blot.Results:(1)The establishment model was succeed.(2)The influnce of RIMM-18 cells on 24h urine protein:At 6d,the 24h urine protein were respectively 10.15±0.47mg/L,10.98±0.85mg/L, 10.92±0.73mg/L of control group,model group and RIMM-18 group.There were not obviously differences among these three groups.At 17d the 24h urine protein were respectively 10.32±0.57mg/L,83.88± 7.88mg/L,78.5±6.44mg/L of control group,model group and RIMM-18 group.The 24h urine protein of model group and RIMM-18 group was obviously higher than control group.But there were not obviously differences between model group and RIMM-18 group.At 31d the 24h urine protein were respectively 10.27±1.21mg/L,150.33±8.8mg/L, 111.68±8.66mg/L of control group,model group and RIMM-18 group.The 24h urine protein of RIMM-18 group was apparently lower than model group.And 24h urine protein of model group and RIMM-18 group were obviously higher than control group.(3) The influnce of RIMM-18 cells on serum albumin:At 6d,the serum albumin were respectively 40.47±1.03g/L,39.05±1.39g/L,37.88±1.81g/L of control group,model group and RIMM-18 group.There were not obviously differences among these three groups.At 17d the serum albumin were respectively 39.83±1.32g/L,21.54±1.86g/L,23.68±1.68g/L of control group,model group and RIMM-18 group.The serum albumin of model group and RIMM-18 group were obviously lower than control group.But there were not obviously differences between model group and RIMM-18 group.At 31d the serum albumin were respectively 38.98±1.41g/L,15.72±1.03g/L,22.13±1.54g/L of control group,model group and RIMM-18 group.The serum albumin of RIMM-18 group was apparently higher than model group.And the serum albumin of model group and RIMM-18 group were obviously lower than control group.(4)The influnce of RIMM-18 cells on serum cholesterol:At 6d,the serum cholesterol were respectively 1.23±0.11 mmol/L,1.33±0.11mmol/L,1.27±0.13 mmol/L of control group,model group and RIMM-18 group.There were not obviously differences among these three groups.At 17d the serum cholesterol were respectively 1.17±0.14mmol/L,6.25±0.43mmol/L, 6±0.23mmol/L of control group,model group and RIMM-18 group.The serum cholesterol of model group and RIMM-18 group was obviously higher than control group.But there were not obviously differences between model group and RIMM-18 group.At 31d the serum cholesterol were respectively 1.2±0.13mmol/L,8.4±0.36mmol/L,5.7±0.36mmol/L of control group,model group and RIMM-18 group.The serum cholesterol of RIMM-18 group was apparently lower than model group.And serum cholesterol of model group and RIMM-18 group were obviously higher than control group.(5)The influnce of RIMM-18 cells on pathological change of renal tissue:There were few pathological changes at renal tissue under light-microscopic.Compared with control group at 6d,mild fusion of foot process could be observed in the model group.At 17d,the fusion was more serious than at 6d when was segmental.At 31d the fusion changed to be extensive-the most serious.There were not also a few differences among these three groups at 6d and 17d.But RIMM-18 group was better than the model group and culture medium group at 31d. (6) The influnce of RIMM-18 cells on the expression of nephrin and podocin which were marker proteins of podocyte:①Immunofluorenscence staining of Nephrin and Podocin scattered following kidney glomerular capillary loops as continuous striation showing green fluorescence in control group.The intensity of immunofluorenscence staining reduced as the passage of time and it was weakest at 31d in model group.There were not also obviously differences between model group and RIMM-18 group at 6d,17d.But the intensity of immunofluorenscence in RIMM-18 group was more powerful than model group at 31d.②Western blotting:At 6d,the expression of Nephrin were respectively 0.74±0.03,0.53±0.04,0.55±0.09 of control group,model group and RIMM-18 group.The expression of Nephrin of model group and RIMM-18 group were obviously lower than control group.At 17d the expression of Nephrin were respectively 0.29±0.05,0.32±0.04 of model group and RIMM-18 group.There were not obviously differences between them.At 31d the expression of Nephrin were respectively 0.73±0.04,0.15±0.01,0.21±0.02 of control group,model group and RIMM-18 group.RIMM-18 group was obviosly higher than model group. But all of them were obviously lower than control group.At 6d,the expression of Podocin were respectively 0.47±0.02,0.28±0.01,0.3±0.02 in control group,model group and RIMM-18 group.The expression of Podocin model group and RIMM-18 group were obviously lower than control group.At 17d the expression of Podocin were respectively 0.18±0.02,0.19±0.01 of model group and RIMM-18 group. There were not obviously differences between them.At 31d the expression of Podocin were respectively 0.48±0.02,0.08±0.01,0.12±0.03 in control group,model group and RIMM-18 group.RIMM-18 group was obviosly higher than model group.But all of them were obviously lower than control group.Conclusions:(1)RIMM-18 could decresed urine protein at AND rats,alleviate foot process fusion.(2)All of these might be realized through the improvement of Nephrin and Podocin which might maintain construction and function of the integrity of SD.
|
PDF Full Text Request