| Objective:Hepatocyte growth factor(HGF) is a pleiotropic polypeptide growth factor. It can stimulate a variety of epithelial and endothelial cells mitosing and moving. It plays an important role in nutrition in tubular morphogenesis, embryonic kidney formation and kidney repair and regeneration. The integrin-linked kinase (ILK) is a newly discovered adhesion protein connecting to integrinβcytoplasmic region. It's a cellular serine/threonine protein kinase. It plays an important role in the regulation of cell cycle, matrix accumulation and tumorigenesis. It also participates in the development and progression of kidney disease. Recent studies show that the higher level of ILK in podocytes is of great significance in maintaining the integrity of the glomerular basement membrane(GBM) and to keep glomerular filtration rate and in preventing leakage of protein. This research observe kidney in small change, minimal(disease mcd) rats from time to time in the level of apolipoprotein Ilk by Histopathology, immunohistochemistry and immunofluorescence, Ilk in the cells of the injury and may further explore the mechanism and interfere, tiny HGF kidney disease model Ilk of expression.Methods:1. Minimal change nephropathy animal model:Clean male Sprogue-Dawley rats 72. They were randomly divided into three groups:control group, model group and hepatocyte growth factor intervention group. The rats of model group and intervention group were injected Adriamycin (concentration of 2mg/ml, dose of 7.5mg/kg) once via vena caud-alis to make minimal change nephropathy animal model. The rats of HGF intervention group were administrated intraperitoneal injection of HGF (30ug·kg-1·d-1) from the third week after injected the Adriamycin until the end of the trial. The rats of control group were injected equivalent saline once via vena caudalis. Killed 6 rats in each group at the 7th,14th,21th, 28th days respectively after modeling and provided kidney samples. The rats were drawn 3ml blood by angular vein before killed for centrifugal separation of serum to test serum albumin,Bun,Cr and Chol level. The rats were collected 24-hour urine with the metabolic cages to measure the 24-hour urine protein by the sulfosalicylic acid method.2.Use the HE, uranyl acetate-lead citrate stain dyeing to observe the renal pathologic abnormalities under light and electron microscopy.3. The localization of ILK in glomerula was detected by immunofluorescence. Anlysis the expression of ILK in each group at different time and the correlation with the degree of podocyte injury.4. Use electrophoresis to detect serum albumin. Use sulfosalicylic acid method to measure the 24-hour urinary protein. Anlysis the changes of serum albumin and urinary protein in three groups and its relation with ILK.Results:1. We succeed duplicating the Rat MCD model.2.Compared with the control group, the expression of ILK in model group reduced on 7th days, but the difference was not statistically significant( p>0.05). The expression of ILK had a marked increase at 14th days and reached maximum on 21th days (p<0.05). Compared with model group, the expression of ILK in intervention group reduced at 21th days and reduced significantly on 28th days (p<0.05).3.Compared with the control group, the urinary protein excretion in model group increased on 7th days and reach its peak at 21th(p<0.01). Compared with model group, the urinary protein excretion in intervention group at 21th days and 28th days decreased significantly (p<0.05).4.Compared with the control group, the serum albumin in model group decreased from the 7th day and reached the lowest on 21 days(p<0.01). Compared with model group, serum albumin in the intervention group were significantly higher at 21 days and 28 days (p<0.05).5. The histomorphology and pathology:In gross appearance, the glomeruler volume in the control and intervention group is normal, dark red, Cortical and Juxtamedullary Nephrons were separated clearly; and the glomeruler volume in the model group is slightly lager, dark red, Cortical and Juxtamedullary Nephrons were also separated clearly. In light microscope, a little glomerular volume slightly increased, inflammatory cell infiltrated, red blood cell leaked in glomerular capillary, fat deg-enerated and lossed in proximal renal tubular, protein casts formatted in renal tubular cavity. And become sever with the time, the 21th day is the most sever. Compared with model group, the glomerular volume in intervention group slightly decreased, red blood cell leaking and inflammatory cell infiltrating decreased, fatty degeneration and protein casts decreased. In electron microscope, we can observe that any basement membranes exposed, some podocyte lossed, the partial foot processes merged or disappeared, the foot processes fusion in model group, but they have eased in the intervention group.6.Correlation analysis showed that the expression of ILK in model group have positive relationship with 24h urinary protein(r= 0.743, P＜0.01).Conclusions: 1The increasing expression and abnormal distribution of ILK may lead to proteinuria and determine podocyte injury as a useful index.2. ILK was positively correlated with proteinuria, the level of ILK in renal tissue have a certain significance with the prognosis of kidney disease.3.HGF can inhibit the expression of glomerular ILK in MCD rats and take protective effect and treatment. |
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