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Background: Diabetic nephropathy is the leading causeof End-Stage Renal Disease (ESRD) emerging indeveloped as well as developing countries, with thecomplicated pathogenesis. The study of expression ofthe genes related to kidney cells e.g. podocytes has beenshown to be associated with the condition, helping in theelucidation of pathogenesis of the disease. Previouslythe gene expression associated was studied in urinesamples. Material and Methods: In the present study, itwas attempted to analyze the mRNA expression ofpodocyte related genes viz. podocalyxin, podocin andsynaptopodin in Peripheral Blood Mononuclear Cells(PBMCs) in patients with diabetes with and withoutnephropathy in comparison with healthy controls byreverse transcriptase Polymerase Chain Reaction(PCR), followed by semi-quantitative PCR. Results:The expression of Synaptopodin (SYNPO) wasincreased in diabetics than the controls, while nosignificant difference was found for Podocalixyn(PODXL) and Podocin (NPHS2). The expression ofPODXL and NPHS2 was significantly up-regulated;SYNPO was unaltered in microalbuminuric patientsthan healthy controls. PODXL and SYNPO wereincreased significantly in nephropathy subjects thancontrols, with no significant change in NPHS2. Theexpression of only PODXL was found to be upregulated in microalbuminuric patients as compared toT2DM patients without nephropathy. PODXL, SYNPOwere significantly up-regulated and NPHS2 wassignificantly down-regulated in nephropathy subjects ascompared to T2DM patients without nephropathy. Asignificant down-regulation was found for NPHS2expression in nephropathy patients than microalbuminuric patients of T2DM with nephropathy.Conclusion: The detection of gene expression of theseproteins can be used as an early marker for the detectionof development of nephropathy in T2DM patients andpreventive measures can be taken to prolong the onsetof nephropathy in these patients, increasing the lifeexpectancy.
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OBJECTIVE: To investigate the relationship between urinary synaptopodin levels and clinical pathology in patients with diabetic nephropathy, in order to provide new biomarkers for diabetic nephropathy. METHODS: Seventy-five patients with diabetic nephropathy confirmed by renal biopsy were enrolled in this study. Those patientsin was in the First Affiliated Hospital of Zhengzhou University and the Second Hospital of Lanzhou University from September 2016 to December 2017. The level of urinary synaptopodin was measured and the relationship between urinary synaptopodin and clinical and pathological changes was analyzed. RESULTS: The levels of urinary synaptopodin/β-Actin in patients with nodular diabetic nephropathy and diffuse sclerosing diabetic nephropathy were significantly higher than those in the early diabetic nephropathy group(P<0.05). Correlation analysis indicated urinary synaptopodin/β-Actin had a positive correlation with 24-hour urine protein(r=0.408, P=0.014), urinary albumin(r=0.0.341, P=0.043) and serum creatinine(r=0.386, P=0.021) CONCLUSION: The level of urinary synaptopodin in patients with diabetic nephropathy is significantly increased and is associated with clinical and pathological conditions, which may become a potential new biomarker for diabetic nephropathy.
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Objective To explore the kidney protection and possible mechanism of Yishen Tongluo Formula (YTF) in rats with membranous nephropathy (MN). Methods A total of 60 SD healthy male rats were randomly divided into 10 for normal group and 50 for MN rat model group. The MN rat model was established by tail iv cationic bovine serum albumin (C-BSA). After successful modeling, they were randomly divided into model group, benazepril group, and YTF groups at low, medium, and high doses (6.61, 13.22, and 26.44 g/kg). Rats in each group were ig administrated once daily for continuous four weeks according to the corresponding dose. At the end of administration, the 24 h urine total protein (UTP), total cholesterol (TC), total protein (TP), albumin (ALB), blood urea nitrogen (BUN), and serum creatinine (Scr) levels were measured. Immunofluorescence was used to detect the deposition of IgG immune complexes in renal tissue. The glomerular basement membrane and podocyte morphology were observed under electron microscope. Immunohistochemistry and qRT-PCR were used to detect the expression of cytoskeleton-related proteins ezrin and synaptopodin in rat kidneys. Results Compared with the model group, the UTP and TC levels in the rats in each treatment group decreased significantly (P < 0.01), and the TP and ALB levels increased significantly (P < 0.01). The middle and high dose groups of YTF were similar to the benazepril group, and the effect was better than the low dose group of YTF. There was no significant difference in the BUN and Scr among the groups. Compared with the control group, the expression of ezrin and synaptopodin mRNA in the kidney of the model group was significantly decreased (P < 0.01). Compared with the model group, the expression of ezrin and synaptopodin mRNA in the podocytes of different treatment groups was increased in different degrees (P < 0.01). The expression levels of ezrin and synaptopodin mRNA in the kidney of rats in the middle and high dose groups of YTF were similar to those in the benazepril group, which were higher than that in the low dose group of YTF. Conclusion YTF has a therapeutic effect on membranous nephropathy in rats. The mechanism may be related to the inhibition of the degradation of podocyte skeleton related proteins ezrin and synaptopodin and the maintenance of the integrity of the podocyte skeleton and foot process.
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BACKGROUND/AIMS: Podocytes play an important role in maintaining the glomerular filtration barrier and in formation of the slit diaphragm. Podocyte loss is associated with chronic kidney disease progression, but it is not clear whether urinary podocyte proteins in urine reflect the clinical extent of glomerular damage. We investigated the correlation between the amounts of urinary podocyte proteins and renal function and albuminuria. METHODS: The study enrolled 33 patients with diabetic kidney disease or glomerular disease and measured urinary podocytes proteins using Western blotting. Urinary podocyte proteins were measured according to the density of the bands on Western blotting. We measured serum creatinine and the spot urine albumin/creatinine ratio as markers of renal damage, and compared the correlation of urinary podocyte protein in the glomerular disease patients. RESULTS: The mean patient age was 49.3 ± 16.5 years, the mean serum creatinine level was 2.30 ± 1.76 mg/dL, and the mean albumin/creatinine ratio was 4.85 ± 3.52. Among the podocyte proteins, urine synaptopodin showed strong correlation with serum creatinine by multivariate regression analysis (p < 0.001) and showed linear correlation (r = 0.429, p < 0.01). Urine podocyte proteins were increased in patients with diabetes, and synaptopodin showed the greatest significant difference (7.68 ± 5.61 vs. 2.56 ± 3.11, p < 0.001), but this might be associated with renal impairment. The urine albumin excretion did not differ between the diabetics and non-diabetics (p = 0.73). CONCLUSIONS: Urine synaptopodin is associated with serum creatinine elevation in the patients with glomerulonephritis including diabetic kidney disease regardless of urine albumin excretion. We suggest that the urine synaptopodin level can predict glomerular damage independently of the urine albumin excretion.
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Humans , Albuminuria , Blotting, Western , Creatinine , Diabetic Nephropathies , Diaphragm , Disease Progression , Glomerular Filtration Barrier , Glomerulonephritis , Podocytes , Proteinuria , Renal Insufficiency, ChronicABSTRACT
objective To investigate the effects of different sections of receptor associated protein (RAP) on the expression and distribution of TRPC6,synaptopodin and podocalyxin in passive Heymann nephritis(PHN). Methods Male Sprague-Dawley rats were injected with three kinds of antisera (anti-RAP full-length serum,anti-RAP N-terminal serum and anti-RAP C-terminal serum)to establish three kinds of PHN models.The control group was injected with normal rabbit serum.The quatitation of 24 h urinary protein,serum albumin and creatinine were taken before injection and one week after PHN model successfully induced.The histopathologic changes of renal tissues were observed by light microscopy.The expression and distribution of TRPC6,synaptopodin and podocalyxin in glomerular podocytes were observed by laser scanning confocal microscopy and analyzed by fluorescence quantitative software after indirect immunofluorescence double staining.Results The quantities of 24 h urinary protein in the three model groups were significantly higher than those of themselves before injection and control groups (P0.05).The expression of TRPC6 in podocytes was higher in the PHN model groups than that of control group.Fluorescence intensity of TRPC6 in RAP full-length group was stronger than that in RAP N-terminal or C-terminal groups.The expressions of synaptopodin and podocalyxin distributed along the glomerular basement membrane as spot,discontinuous short line and defect of some segments,and were lower in three PHN groups than those of control group.Fluorescence intensity of synaptopodin and podocalyxin among three PHN groups had no differences. Conclusions RAP full-length and N-terminal or C-terminal parts can increase the expression of podocyte TRPC6,but decrease the expressions of synaptopodin and podocalyxin,and alter their distribution,which may be associated with the proteinuria,however,their role in the PHN pathogenesis needs further study.