Evaluation of Podocalyxin level in pre-eclampsia with severe features' patients: a cross-sectional study

Background: This study aims to evaluate the level of podocalyxin (PCX) in preeclampsia with severe features patients and correlate it with the results of laboratory tests.Methods: The current study was a cross-sectional study conducted in Assiut Women Health Hospital between April and October 2018.  The study included 60 patients divided into two groups; Group (A): 30 patients diagnosed to have preeclampsia with severe features and Group (B): 30 patients as normal control group. Complete laboratory investigations with measurements of the PCX level was performed for all study participants.Results: No statistically significant difference between the study group and control group according to blood urea (p= 0.339) and serum creatinine (p= 0.801).There was statistically significant difference between the study group and control group according to PCX level (p= 0.001); the mean PCX was 3340.0 ± 2394.6 in the study group versus 1083.5±1400.2 in the control group. Univariate analysis revealed podocalyxin was not correlated with clinical data or laboratory investigations.Conclusions: Podocalyxin levels were significantly elevated in preeclampsia.

Value of podocalyxin levels in urinary extracellular vesicles for diagnosis of diabetic nephropathy / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the value of detecting podocalyxin (PCX) level in urinary extracellular vesicles for the diagnosis of diabetic nephropathy.</p><p><b>METHODS</b>This study was conducted among 57 diabetic patients admitted during the period from March to September, 2017, including 34 with uncomplicated diabetics and 23 with diabetic nephropathy; 21 patients with other types of nephropathy and 11 healthy individuals were also included to serve as the controls. Transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA) were used to verify the separation of urinary extracellular vesicles. The molecular markers of extracellular vesicles (TSG101 and podocalyxin [PCX]) were detected using Western blotting. PCX levels in extracellular vesicles were also detected using ELISA.</p><p><b>RESULTS</b>TEM reveal the presence of numerous extracellular vesicles in the urine with intact morphology and different sizes, and most of them were below 300 nm in diameter as shown by NTA. TSG101 expression was detected in the samples from all the 4 groups. Positive expression of PCX was detected in the samples from patients with diabetic nephropathy but not in the other groups. In patients with diabetic nephropathy, the mean PCX levels (3.27±2.30 ng/μmol)was significantly higher than those in the healthy control group (1.22±0.36 ng/μmol), uncomplicated diabetes group (2.22±1.29 ng/μmol) and nephropathy group (1.24±0.45 ng/μmol).</p><p><b>CONCLUSIONS</b>PCX level in urinary extracellular vesicles is significantly increased in patients with diabetic nephropathy, suggesting the value of PCX as a potential marker for clinical diagnosis of diabetic nephropathy.</p>

Diagnostic values of serum free fatty acids and urine podocalyxin in the patients with type 2 diabetic nephropathy / 临床检验杂志

Objective To investigate the diagnostic values of serum free fatty acids (FFA) and urine podocalyxin (PCX) in the patients with type 2 diabetic nephropathy(DN).Methods A total of 120 patients with type 2 diabetes mellitus (DM) were enrolled,and they were divided into three groups,normal albumin group(NA),microalbuminuria group(MA) and heavy albuminuria group(HA),based on the urinary albumin-creatinine ratio(UACR).In addition,40 healthy persons participated in medical examination were selected as the control group.Serum FFA levels were detected by a biochemical analyzer and urine PCX levels by ELISA.The correlation between them was analyzed by Pearson correlation,and their diagnostic values in type 2 DN were evaluated by the receiver operating characteristic curve (ROC).Results Serum FFA levels in the NA,MA,HA and control groups were (0.61 ± 0.14),(0.81 ± 0.13),(0.95 ± 0.18) and (0.49 ± 0.11) mmol/L,respectively,and urine PCX levels were (1.86 ± 0.45),(4.47 ± 1.48),(6.72 ± 1.40)and(1.38 ±0.24) ng/mL,respectively.The levels of serum FFA and urine PCX in type 2 DM patients were significantly higher than those in the control group(P ＜ 0.05),and increased with the progression of type 2 DM.Pearson correlation analysis showed that serum FFA levels were positively correlated with urine PCX levels(r =0.73,P ＜ 0.05).The sensitivity of serum FFA,urine PCX and combined detection in the diagnosis of type 2 DN were 74.1％,80.6％ and 89.5％,respectively,and the latter significantly higher than the former two (P ＜ 0.05).Conclusion Serum FFA and urine PCX levels increase significantly in diabetic patients with renal injury,and both of them have important clinical values in the diagnosis of type 2 DN.

Clinical observation of Shenluoan Decoction for treating obese patients with early diabetic nephropathy / 中草药

Objective: To observe the therapeutic effects and possible mechanism of Shenluoan Decoction on the obese patients with early diabetic nephropathy. Methods: Obese patients with early diabetic nephropathy (106 cases) were randomly assigned into control group (n = 53, treated by Irbesartan) and treatment group (n = 53, treated by Shenluoan Decoction combined with Irbesartan), and the conventional Western therapy was given to all the patients in both groups. The changes of therapeutic efficacy, Chinese medicine syndrome, such as waistline, serum creatinine (SCr), blood urea nitrogen (BUN), total cholesterol (TC), triglyeride (TG), fasting blood glucose (FBG), urinary podocalyxin (PCX), urinary nephrin, urinary albumin, and urine creatinine were detected before and after 6 months treatment. Results: The experiment was completed with 102 cases. The total effective rate in the treatment group (90.00%) was significantly higher than that of control group (67.31%) (P 0.05). After the treatment, the levels of SCr, TC, and TG were reduced significantly in the treatment group (P < 0.05), and showed significant difference compared with those in the control group (P < 0.05). The levels of UPCX/UCR, UNephrin/UCR, and UACR were decreased with significant differences compared with those before treatment and in the control group, respectively (P < 0.05). UPCX/UCR, UNephrin/UCR, and UACR showed positive correlation. Conclusion: The combination of Shenluoan Decoction and Irbesartan has a better effect than the single use of Irbesartan on the obese patients with early diabetic nephropathy. Its mechanism may be related to providing some renal protection and reducing the PCX and nephrin excretion.

Effect of Nocturnal Hypertension in Primary Hypertension Patients With Early Renal Dysfunction / 中国循环杂志

Objective: To investigate the effect of nocturnal hypertension in essential hypertension (EH) patients with early renal dysfunction. Methods: A total of 182 EH patients were enrolled in this study. According to weather the average night blood pressure (BP) > 120/70 mmHg, the patients were divided into 2 groups: Nocturnal hypertension group,n=89 and Control group,n=93. The levels of urine micro albumin (mALB), podocalyxin (PCX) and serum Cyst-C were examined and compared between 2 groups. Results:① For BP: the average systolic and diastolic BP in Nocturnal hypertension group were higher than those in Control group as SBP (138.05 ± 6.33) mmHg vs (102.51 ± 8.76) mmHg, DBP (84.11 ± 6.32) mmHg vs (70.03 ± 4.56) mmHg, P<0.05. ② For renal dysfunction biomarkers: several biomarker levels were higher in Nocturnal hypertension group than those in Control group as mALB (13.60 ± 0.69) mg vs (10.04 ± 0.73) mg, PCX (5.35 ± 1.69) ng/ml vs (2.05 ± 0.88) ng/ml and serum Cyst-C (1.35 ± 0.69) mg/L vs (1.02 ± 0.44) mg/L, allP<0.05.③ For correlation study: In Nocturnal hypertension group, the PCX level was positively correlated to the levels of mALB (r=0.675,P<0.05), Cyst-C (r=0.734,P<0.05), night BP (r=0.830, P<0.05) and the duration EH (r=0.688,P<0.05). Conclusion: EH patients with nocturnal hypertension have more incidences to suffer from renal dysfunction, the examination of mALB, PCX and Cyst-C are beneifciary for the early diagnosis in relevant patients.

Hydrochloride pioglitazone protects diabetic rats against podocyte injury through preserving glomerular podocalyxin expression / O cloridrato de pioglitazona protege ratos diabéticos contra a injúria aos podócitos por meio da preservação da expressão de podocalixina glomerular

Objective: We sought to test the effect of different dosages of pioglitazone (PIO) on the glomerular expression of podocalyxin and urinary sediment podocalyxin excretion and to explore the potential renoprotective mechanism. Materials and methods: Type 1 diabetes induced with streptozotocin (65 mg/kg) in 36 male Sprague-Dawley rats were randomly allocated to be treated with vehicle or 10, 20, 30 mg/kg/d PIO respectively for 8 weeks. Eight rats were enrolled in the normal control group. Results: At 8th week, rats were sacrificed for the observation of kidney injury through electron microscope. Glomerular podocalyxin production including mRNA and protein were determined by RT-PCR and immunohistochemistry respectively. Levels of urinary albumin excretion and urinary sediment podocalyxin, kidney injury index were all significantly increased, whereas expression of glomerular podocalyxin protein and mRNA were decreased significantly in diabetic rats compared to normal control. Dosages-dependent analysis revealed that protective effect of PIO ameliorated the physiopathological changes and reached a peak at dosage of 20 mg/kg/d. Conclusion: PIO could alleviate diabetic kidney injury in a dose-dependent pattern and the role may be associated with restraining urinary sediment podocalyxin excretion and preserving the glomerular podocalyxin expression. .

Objetivo: Buscamos testar os efeitos de diferentes doses de pioglitazona (PIO) sobre a expressão glomerular de podocalixina e sobre a excreção de podocalixina em células do sedimento urinário, além de explorar o potencial mecanismo de proteção renal. Materiais e métodos: O diabetes tipo 1 foi induzido em 36 ratos Sprague-Dawley machos com estreptozotocina (65 mg/kg). Os animais foram tratados apenas com o veículo, ou com 10, 20, 30 mg/kg/d de PIO por 8 semanas. Oito ratos foram colocados no grupo controle. Resultados: Na oitava semana, os ratos foram sacrificados para se observar a lesão renal em microscopia eletrônica. A produção de podocalixina glomerular, incluindo mRNA e proteína, foi determinada por RT-PCR e imuno-histoquímica, respectivamente. Os níveis urinários de albumina e podocalixina nas células do sedimento urinário e o índice de lesão renal estavam todos significativamente aumentados, enquanto a expressão glomerular da proteína podocalixina e do mRNA estava significativamente diminuída em ratos diabéticos comparados com o controle normal. A análise dos efeitos dose-dependentes revelou que o efeito protetor da PIO melhorou as mudanças fisiopatológicas e atingiu um pico na dose de 20 mg/kg/dia. Conclusão: A PIO pode melhorar a injúria renal de forma dose-dependente e este papel pode estar associado com a prevenção da excreção de podocalixina nas células do sedimento urinário e com a preservação da expressão glomerular de podocalixina. .

Role of PTEN in podocyte injury in patients with diabetic nephropathy / 临床与实验病理学杂志

Purpose To investigate the role of PTEN in podocyte injury in patients with diabetic nephropathy ( DN) . Methods Uri-nary samples from 30 patients with DN and 10 healthy volunteers were collected to detect the level of PCX by ELISA. Renal biopsies were reviewed to observe the morphological changes. All patients with DN were divided into three groups by glomerular lesion. The ex-pression of p-Akt and PTEN in glomeruli was detected by immunohistochemistry. Results The levels of PCX in the urine were signifi-cantly higher in patients with DN compared with those in healthy volunteers, and gradually increased along with glomerular lesion aggra-vating. The expression of p-Akt and PTEN increased in patients with DN compared with healthy volunteers. Although the expression of p-Akt and PTEN decreased with the aggravation of glomerular lesion, they were still higher than that in volunteers. There were obvious-ly positive correlation between the level of PCX and 24-h urinary protein and negative correlation between the level of PCX and the ex-pression level of p-Akt and PTEN. Conclusion PTEN down-regulation may be associated with podocyte injury in DN, which may be associated with the phosphorylation of Akt.

The expression of podocalyxin in patients with diabetic nephropathy and its clinical significance / 中国医师进修杂志

Objective To investigate the expression ofpodocalyxin(PCX) in patients with diabetic nephropathy (DN) and its clinical significance.Methods A total of 68 patients with DN were enrolled as DN group.Another 20 healthy person were selected as control group.The patients with DN were divided into three sub-groups:normal albuminuria group(＜ 30 mg/g,group A),microalbuminuria group(≥30 mg/g but ＜ 300 mg/g,group B) and macroalbuminuria group (≥ 300 mg/g,group C).The level of urinary PCX was measured with enzyme-linked immunosorbent assay.Results The level of urinary PCX,serum creatinine and blood urea nitrogen in DN group was higher than that in control group [(27.44 + 6.87) μ g/L vs.(3.52 +1.13) μ g/L,(154.82 ± 11.54) μ mol/L vs.(82.56 ± 5.71) μ mol/L,(8.79 ± 0.58) mmol/L vs.(4.23 ± 0.34)mmol/L,P ＜0.01].The level of urinary PCX in group A,B,C was (18.04 ±4.12),(23.58 ±5.43),(32.18 ± 6.05) μ g/L,and there was significant difference among three groups (P ＜ 0.05).The level of serum creatinine and blood urea nitrogen was (138.54 + 8.66),(142.16 ± 11.74),(165.13 ± 12.85) μ mol/L and (8.48 ± 0.43),(8.56 ± 0.61),(10.04 + 1.17) mmol/L in group A,B,C,there was no significant difference between group A and group B(P＞ 0.05),but there was significant difference between group A and group C,or between group B and group C (P ＜ 0.05).Conclusion The high-level expression of urinary PCX is found in DN patients,which maybe play a key role in the early diagnosis and assessment of the progression of DN.

Podocalyxin expression in renal tissues and correlation with the number of urinary podocytes in children with Henoch-Sch(o)nlein purpura nephritis / 中南大学学报(医学版)

Objective:To analyze the podocalyxin (PCX) expression in the kidney and the number of urinary podocytes in different pathological grades of Henoch-Sch(o)nlein purpura nephritis (HSPN),and to determine whether the number of urinary podocytes reflects the renal damage in HSPN.Methods:Fifty-six children diagnosed with HSPN in our hospital were enrolled in the study and classified into 4 groups by renal pathology:grade Ⅱ (Ⅱa+Ⅱb) (n=10),grade Ⅲ (Ⅲa+Ⅲb) (n=21),grade Ⅳ (n=16),and grade Ⅴ (n=9).Four kidney autopsy specimens without histomorphologic lesions and 8 urine samples from healthy children served as controls.With immunofluorescence assay,the PCX expression in 4 normal renal tissues and in the renal tissues of the 56 HSPN children was detected and quantitatively analyzed.Positive rate and the number of urinary podocytes were detected in the 8 healthy children and 56 HSPN children.Results:In the renal tissues of the normal control group and grade Ⅱ (Ⅱa+Ⅱb) HSPN group,the PCX expression was complete.The percentage of the PCX positive area out of the total glomerular area in the renal tissues of 2 groups had no significant difference (P＞0.05).In the renal tissues of grade Ⅲ (Ⅲa+Ⅲb),Ⅳ,and Ⅴ HSPN groups,the PCX expression showed various degrees of loss,decreasing in turn from grade Ⅱ (Ⅱa+Ⅱb),Ⅲ (Ⅲa+Ⅲb),Ⅳ to Ⅴ,with significant differences between each group (P＜0.01).For HSPN with grade Ⅲ (Ⅲa+Ⅲb) or higher,positive PCX expression was found in the urine,suggesting the presence of enough podocytes in the urine.The percentage of fluorescence positive area out of the total glomerular area of PCX in the renal tissues was negatively correlated with the total number of urinary podocytes (r=-0.637,P＜0.01).Conclusion:Podocyte injury plays a certain role in the pathological progression of HSPN.The urinary detection ofpodocytes can reflect the degrees of pathological damage in HSPN.

Podocalyxin in podocyte injury of proteinuria associated nephropathy / 国际儿科学杂志

Podocalyxin,a sialomucin most closely related to CD34,is expressed by kidney podocytes,hematopoietic progenitors,vascular endóthelia and a subset of neurons.As one of the important apical marker protein,podocalyxin plays an important role in the integrity of the structure and function of nomal podocyte,and is closely related to the prevention of proteinuria.The mechanism of podocyte injury associated with proteinuria remains unkown.The investigation of podocalyxin,a major podocyte protein,is significant in the pathogenesis and process of the primarily glomerulopathies.This paper summarizes the progresses of podocalyxin in podocyte injury of proteinuria associated nephropathy.

Effects of the different sections of receptor-associated protein on the expression and distribution of TRPC6,synaptopodin and podocalyxin in passive Heymann nephritis / 中华肾脏病杂志

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.

Intervention of azithromycin on adriamycin-induced nephropathy rats / 中华肾脏病杂志

Objective To investigate the effects of azithromycin on serum indicators, urine indicators, renal pathology, intercellular adhesion molecule-1 (ICAM-1), nephrin and podocalyxin in adriamycin(ADR)-induced nephropathy(ADN) rats. Methods ADR nephropathy was induced by a tail intravenous injection of ADR. The rats were randomly divided into azithromycin-treated group, prednisone -treated group, integrated treatment group, model group and control group. Serum index and 24 h urine protein were measured serially at 0, 4th, 8th weeks. The values of creatinine clearance (Ccr) were calculated. Kidney tissues were collected for microscopy observation. The expressions of nephrin, podocalyxin and ICAM-1 in renal tissue were detected by immunohistochemistry SP. Results Compared with normal control group, at the 4th week, 24 hours urine protein and albumin reached the level of ADR nephropathy model. Compared with model group, at the 8th week, 24 h urine protein, cholesterol, serum creatinine, renal pathology changes in the three treated group rats were significantly reduced (P<0.05), serum total protein, albumin and Ccr (except C group) significantly raised (P<0.05), ECM/GA and renal pathology score significantly reduced (P<0.01), ICAM-1 significantly decreased (P<0.01), nephrin and podocalyxin significantly increased. Besides, the curative effect of integrated treatment was better than other treatment group. Conclusions Azithromycin induces similar responses in ADR nephropathy as prednisone, but its early protective effect of renal function is worse than prednisone.The integrated treatment of azithromycin and prednisone has a synergistic effect, and the efficacy is superior to each drug alone.

Correlation of urinary podocyte number and glomerular podocalyxin expression with clinicopathology in IgA nephropathy patients / 中华肾脏病杂志

Objective To examine the correlation of urinary podocyte number and giomerular podocalyxin expression with clinicopathology in IgA nephropathy(IgAN)patients. Methods Morning urinary specimens(100 ml)3 days before renal biopsy from 50 patients with IgAN diagnosed by renal biopsy and from 20 healthy volunteers as control were collected. After centrifugation, 300 μI sediment was used for smear. Immunohistochemical staining with monoclonal anti-podocalyxin antibody was performed to detect urinary podocytes and the number of podocyte was counted under optical microscope. Computer image analysis system was used to examine glomerular PCX expression. Renal pathology and classification were investigated based on Lee's grading and Katafuchi semi-quantitative integration method. Relevance analysis was carried out on urinary podocyte number, glomerular PCX expression with pathological score and clinical data. Results The amount of urinary podocytes in IgAN was obviously higher than that in healthy controls(P<0.01). Significant differences were found in multiple comparison of the median of urinary podocytes among Lee's grade groups. I - II group was lower as compared to Ⅲ , Ⅳ, Ⅴ groups(all P<0.05). Ⅲ group was lower as compared to V group(P<0.05). The positive rate of urinary podocyte was the highest in Ⅳ and V groups(100%), while the lowest in Ⅰ - Ⅱ group(55%). Glomerular PCX expression in IgAN decreased with the aggravation of renal pathology. Significant differences were found in multiple comparison of the glomerular PCX expression with the pathological score. Lee's Ⅰ - Ⅱ group was higher as compared to Ⅲ, Ⅳ, Ⅴ groups(all P<0.05). Ⅳ and Ⅳ groups were higher as compared to V group(P<0.05). In IgAN, urinary podocyte excretion was negatively correlated with glomerular PCX expression(r=-0.702, P<0.01), positively correlated with 24-hour urinary protein(r=0.465, P<0.01)and positively correlated with glomerular and tubular scores(r=0.233, 0.307, P<0.05). Glomerular PCX expression was negatively correlated with 24-hour urinary protein(r=-0.367,P<0.05)and negatively correlated with glomerular and tubular scores(r =-0.560, -0.377, P <0.05). Conclusions Injury and desquamation of glomerular podocytes may involve in the development of IgAN. The number of urinary podocyte can reflect the loss of podocytes in renal tissue, which may be used as a marker of disease progression of IgAN.

Effect of sulodexide on podocalyxin expression of podocytes in streptozotocin diabetic desoxycorticosterone acetate-hypertensive rats / 中华肾脏病杂志

Objective To explore the effect of suledexide on renal injury and podocalyxin expression of podocytes in STZ diabetic desoxycorticosterone acetate (DOCA)-hypertensive rats. Methods Wistar rats were subjected to subcutaneous injection of streptozotocin(STZ), followed by uninephrectomy and subcutaneous administration of DOCA. Diabetic and hypertensive rats were randomly allocated to treatment with sulodexide or a combination of sulodexide and telmisartan for 8 weeks. Blood pressure (BP), 24-hour urinary albumin were measured every 2 weeks. Blood and urinary samples were collected to detect biochemical indexes of plasma and urinary β-acetyl-β-D-glucosaminidase (NAG) at the end of the study. Immunohistochemistry (IHC), RT-PCR and Western blot were performed to examine the expression and distribution of podocalyxin. Results STZ +DOCA-treated rats progressively developed hypertension, albuminuria and hyperglycemia. Hyperlipidemia and hypoinsulinemia were found in diabetic and hypertensive rats compared with controls. Albuminuia was significantly reduced in sulodexide group at week 8 and sulodexide plus telmisartan group at week 6 and week 8. Blood pressure decreased in sulodexide plus telmisartan group. No significant effects on lipid and glucose metabolism were observed in all treated groups. Histopathological index increased in STZ+DOCA-treated rats, but was significantly lower in sulodexide group as well as sulodexide plus telmisartan group. The number of podocytes on glomerular cross-section of the four groups were comparable. Segmental loss and down-regulation of podocalyxin were detected in STZ+DOCA-treated rats, which were greatly attenuated by sulodexinde, meanwhile, combination treatment preserved more podocalyxin expression in glomeruli than sulodexide monotherapy. Conclusion Sulodexide effectively reduces albuminuria, prevents loss of podocyte podocalyxin and alleviates renal damage in STZ diabetic DOCA-hypertensive rats.