Chinese Journal of Nephrology

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Mesangial cell-derived tumor necrosis factor α up-regulates the expression of tubular liver-type fatty acid binding-protein and its renoprotective role in IgA nephropathy

ZUO Nan;LI Xiao-li;WANG Li-ning;LI Zi-long;WANG Jun;FENG　Jiang-min;MA Jian-fei;FAN Qiu-ling;YAO Li.

2011, 27(7): 469-474.

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Objective To explore the mechanism of up-regulation of tubular liver-type fatty acid binding-protein (L-FABP) in IgA nephropathy (IgAN) and its renoprotective role. Methods Murine mesangial cells (MCs) from primary cell culture were cultured with aggregated IgA (AIgA) (10 to 250 mg/L) for 48 hours. The supernatant after culture was collected as AIgA-MC medium. Murine proximal tubular cell line (mProx) stably expressing human L-FABP (hL-FABP) by transfection (mProx-L) were cultured with AIgA, AIgA-MC medium and /or neutralizing anti-TNF-α antibody and recombinant murine TNF-α, respectively. AIgA-MC medium (AIgA final concentration was 25 mg/L) was cultured with mProx and mProx-L cells. The mRNA expressions of hL-FABP and MCP-1 of the cells were detected by real-time PCR. The protein expressions of hL-FABP and 4-HNE of the cells were detected by Western blotting. Results (1) The hL-FABP mRNA and protein expression stimulated by AIgA-MC medium was significantly higher as compared to AIgA (P<0.01). (2) Pre-incubation of neutralizing anti-TNF-α antibody （final concentration was 1 and 5 mg/L) with mProx-L cells could significantly suppress the up-regulation of hL-FABP protein expression induced by AIgA-MC medium (P<0.05 and P<0.01). (3) Recombinant murine TNF-α (final concentration was 50 and 250 ng/L) also induced a significant up-regulation of hL-FABP expression (P<0.01). (4) After the stimulation of AIgA-MC medium, both 4-HNE protein expression and MCP-1 mRNA expression were significantly suppressed in mProx-L cells compared to those of mProx cells (P<0.05 and P<0.01). Conclusion Mesangial cell-derived TNF-α can induce up-regulation of tubular L-FABP expression. Over-expression of tubular L-FABP may lessen the progression of IgAN by reducing oxidative stress and inflammatory mediators.

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Expression of B1a cells and IgA1 positive cells in tonsil of IgA nephropathy patients and analysis of associated clinicopathological factors

HOU Qi-di;WU Gang;PENG You-ming;LIU Hong;LIU Ying-hong;XU Xiang-qing;JIANG Wen-ling;LIU Fu-you.

2011, 27(7): 475-478.

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Objective To examine the expression of IgA1 and B1a positive cells in palatine tonsils of IgA nephropathy (IgAN) patients, and to analyze the association between B1a cells and clinicopathological changes. Methods Eight patients diagnosed as IgAN by renal biopsy and 8 chronic tonsillitis patients without nephritis as control were enrolled in the study. Immunofluorescence and laser scanning confocal microscope (LSCM) were applied to observe the localization and quantitative calculation of B1a and IgA1 positive cells. Statistic analysis of the association of B1a cells with proteinuria and pathological Lee’s grading was performed. Results B1a cells were mainly localized in germinal center of tonsil, and IgA1 positive cells were mainly localized in subepithelium of tonsil. Compared to control group, the percent of B1a cells and IgA1 positive cells was significantly higher in IgAN (P<0.01). There was a positive correlation between B1a cells and IgA1 cells (P<0.05). In IgAN, the percent of B1a cells in patients with hematuria and proteinuria was obviously higher than that of patients with hematuria only (P<0.05). The number of B1a cells in IgAN patients with ≥Lee's grade Ⅲ was significantly higher than that of those < grade Ⅲ (P<0.05). Conclusions IgA1 may be secreted by B1a cells in the tonsil of IgAN patients. The number of B1a cells is correlated with exacerbation of proteinuria and pathological severity, which may play an important role in pathogenesis of IgAN.

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Expression of response gene to complement 32 in renal tissue of children with IgA nephropathy and its significance

NIU Xiao-ling;KUANG Xin-yu;ZHANG Zhi-gang;LIU Xue-guang;ZHAO Zhong-hua;ZHANG Xin;XU Hong;HUANG Wen-yan.

2011, 27(7): 479-483.

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Objective To examine the expression of response gene to complement 32(RGC-32) in renal tissue of children with IgA nephropathy(IgAN), and to explore its significance. Methods The subjects were 45 children diagnosed as IgAN by renal biopsy. The expression of RGC-32, α-smooth muscle actin (α-SMA) and transforming growth factor β1(TGF-β1) was examined by immunohistochemistry staining. The correlation of RGC-32 expression with α-SMA, TGF-β1, degree of renal pathological lesions and clinical index in IgAN was assessed by Spearman correlation analysis. Results RGC-32 protein located in renal tubular epithelial cells in normal and IgAN renal tissues. The positive expression index of RGC-32 in nomal group, IgAN mild group, moderate group and severe group was (18.29±6.22)%, (23.90±9.65)%, (31.23±9.86)%, and (34.52±10.63)% respectively. With more severity of renal pathological lesions, the expression of RGC-32 in IgAN was enhanced. The RGC-32 expression was positively correlated with the score of glomerulus and renal interstitium in children with IgAN (r=0.385, 0.347, P<0.05), as well as α-SMA, TGF-β1 (r=0.594, 0.521, P<0.01), but was not correlated with Scr, urinary NAG/Cr, Alb/Cr, IgG/Cr, and α1-M/Cr (r=0.117, -0.115, -0.138, -0.176, -0.028, all P>0.05). Conclusions RGC-32 protein locates in renal tubular epithelial cells in normal and IgAN renal tissues. RGC-32 may participate in the course of renal tubulointerstitial lesions in children with IgAN, especially in the course of epithelial-mesenchymal transition (EMT) induced by TGF-β1.

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Association between Mycoplasma penetrans infection and clinicopathology of IgA nephropathy

JIANG Xue;XU Fei-fei;ZHANG Hui-di;ZHANG Jian-na;SHI Yan-ling;SUN Mei.

2011, 27(7): 484-487.

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Objective To study the association between Mycoplasma penetrans (Mpe) infection and clinicopathology of IgA nephropathy (IgAN). Methods Blood samples of 118 IgAN patients, 90 patients with chronic kidney disease (CKD) and 89 healthy people were collected. Mpe DNA in serum was detected by PCR and positive samples were confirmed by Southern blotting. According to Mpe infection, IgAN patients were divided into positive and negative groups. Association between clinicopatholgical features of IgAN and Mpe infection was examined. Results Significantly higher Mpe positive rate was found in IgAN group as compared to CKD and healthy groups (16.0% vs 2.2% and 1.1%, P＜0.01). In Mpe positive group, 42.1% patients presented macroscopic hematuria, which was significantly higher than that in Mpe negative group (P＜0.01). While Mpe negative group had greater proteinuria, higher serum creatinine level, higher Lee grading of pathology compared to Mpe positive group. There were no differences of tubulointerstitial lesions and arteriole hypertrophy between two groups. Conclusions IgAN patients have higher Mpe infection rate than CKD patients and healthy people. Mpe positive IgAN patients have more macroscopic hematuria. Mpe infection may be associated with the pathogenesis of IgAN.

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Expression of intrarenal angiotensin Ⅱ is correlated with kidney fibrosis in primary IgA nephropathy patients

ZHANG Xiao-yan;LV Wen-lv;TENG Jie;ZHONG Yi-hong;DING Xiao-qiang.

2011, 27(7): 488-491.

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Objective To analyze the expression and regulation of components of intrarenal renin-angiotensin system (RAS) and the correlation between intrarenal angiotensinⅡ(Ang Ⅱ) expression and clinicopathological injury index in primary IgA nephropathy patients. Methods Expressions of intrarenal RAS components were assessed by immunohistochemistry staining (IHCS). Correlation among intrarenal RAS components and of intrarenal Ang Ⅱ expression with blood pressure, estimated glomerular filtration rate (eGFR), 24-h urinary protein and Katafuchi score in 36 primary IgA nephropathy patients were examined. Results There were positive correlations between positive IHCS area of intrarenal renin and AngⅡ(r＝0.43, P<0.01), angiotensiongen and AngⅡ (r＝0.34, P<0.05). There was negative correlation between positive IHCS area of intrarenal AngⅡ and eGFR(r＝-0.61, P<0.01). There was positive correlation between positive IHCS area of intrarenal AngⅡ and pathological chronicity index (ρ＝0.39, P<0.05), index of interstitial cell infiltration (ρ＝0.52, P<0.05). Conclusion Expression of intrarenal AngⅡ is positively correlated with expression of intrarenal renin and angiotensinogen, and plays an important role in kidney fibrosis in primary IgA nephropathy.

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ost-hemodialytic urea rebound and its impact factors in maintenance hemodialysis patients

LU Jia;XU Shao-wei;DING Xiao-qiang;ZOU Jian-zhou;LIU Zhong-hua;SHEN Bo;TENG Jie.

2011, 27(7): 495-498.

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Objective To study the urea rebound after hemodialysis in maintenance hemodialysis (MHD) patients and its impact factors. Methods From 124 stable MHD patients, blood samples were collected at the beginning, immediate post-hemodialysis, 15 minutes and 30 minutes after hemodialysis. The urea rebound was quantified, and its effect on URR and spKt/V was investigated. The impact factors on urea rebound were analyzed. Results In this group of patients, average post-hemodialytic urea rebound was 13.6%, leading to over-estimation of URR and spKt/V of 0.04 and 0.14, respectively. Hemodialysis efficiency expressed as K/V determined urea rebound most significantly. Other impact factors included higher hemoglobin, higher relative ultrafiltration, arteriovenous access, and male patients. Conclusions Urea rebound is common after the hemodialysis. For specific patients and hemodialysis sessions, ignoring it would result in significant over-estimation of delivered hemodialysis dose.

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Effect of calf bioimpedance ratio for volume evaluation on hypertension in hemodialysis patients

LIU Jing;ZHOU Yi-lun;SUN Fang;MA Li-jie;SHEN Yang;JIANG Wei;CUI Tai-gen.

2011, 27(7): 499-503.

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Objective To elucidate the effect of bioimpedance ratio in the calf (calf-RBI) for volume evaluation on hypertension in hemodialysis (HD) patients. Methods Bioimpedance in the right calf was measured by bioimpedance spectroscopy (BIS). As an index of volume status, calf-RBI was calculated as follows: calf-RBI =impedance at 200 kHz / impedance at 5 kHz. The range of age-stratified 1SD from mean calf-RBI in the healthy control was assumed as the target range for the corresponding HD patients. The dry body weight (DBW) was stepwise decreased under the guidance of calf-RBI. The changes of calf-RBI, blood pressure and antihypertensive medications were recorded and correlation analysis among indexes was performed. Results The calf-RBI showed a normal distribution in both healthy subjects and HD patients. The calf-RBI was positively correlated with age, but not with gender or BMI. Forty-two patients with (35.9%) calf-RBI beyond target range were identified in 117 HD patients. The percentage of uncontrolled hypertensive individuals was significantly higher as compared to those with calf-RBI within or below target range (59.5% vs 33.3% and 16.7%, P＜0.01). The percentage of uncontrolled hypertensive individuals and the dose of antihypertensive medications was significant improved after decreasing the DBW in the patients with calf-RBI beyond target range (74.1% vs 33.3%, P＜0.01) and defined daily dose (2.00±2.28 vs 2.49±2.47, P＜0.05 ). Conclusions The age-stratified calf-RBI may be used as a useful index for estimation of volume status, and has a good association with clinical manifestations. Recognition and correction of chronic fluid overload based on age-stratified calf-RBI is helpful in hypertension control for hemodialysis patients.

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Epidemiology investigation of chronic kidney disease in adults of Shanghai Pudong New Area

ZHOU Yi*;SUN Qiao;RUAN Xiao-nan;XU Wang-hong;ZHAO Gen-ming;GU Jian-jun;YANG Li-ming;FU Xiao-jin;BAI Yun;ZHANG Hong;QIU Hua.

2011, 27(7): 504-510.

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Objective To investigate the prevalence and risk factors of chronic kidney disease (CKD) in adult residents of Shanghai Pudong New Area. Methods A total of 5584 residents aged 20-80 years old were randomly selected from Shanghai Pudong New Area through multistage sampling and interview between April and July 2008. Fasting blood samples and morning urine samples were collected to detect Scr and urinary albumin and creatinine. Urine albumin to creatinine ratio (ACR) and glomerular filtration rate (GFR) was calculated to estimate the renal function. Logistic regression model was used to examine the associations of demographic and lifestyle factors with indicators of kidney damage. Results The age-standardized prevalence of albuminuria, reduced renal function and CKD was 9.9% (male 8.0%, female 12.4%), 1.1% (male 1.3%, female 0.9%) and 11.0%(male 8.8%, female 12.7%), respectively. The prevalence of CKD was higher in female and increased with age. No significant difference in the prevalence was observed between urban and rural areas. Elder, female, high blood pressure, high fasting glucose, dyslipidemia and obesity were associated with CKD independently. Conclusions The prevalence of CKD in Shanghai Pudong New Area is comparable to that previously reported in China or other developed countries, and even higher. CKD is going to be a public health problem and warrants the community-integrated control strategy to prevent the incidence of end-stage renal disease (ESRD) and related complications.

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Anti-erythropoietin antibody level in maintenance dialysis patients and its clinical significance

PENG Zhi-yuan;ZHANG Yi-ling;ZHAO Jiu-yang.

2011, 27(7): 511-514.

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Objective To investigate the anti-erythropoietin antibody level and its clinical significance in maintenance dialysis patients. Methods Eighty maintenance hemodialysis (HD) and 30 peritoneal dialysis (PD) patients were enrolled in the study. Serum anti-erythropoietin antibody levels of above 110 dialysis patients were measured by ELISA. Immunoreactive parathyroid hormone (iPTH), Scr, BUN, Hb, and CRP were determined by conventional methods at the same time. Correlations among these indexes were examined. Results The anti-erythropoietin antibody levels of the dialysis patients were significantly higher than those of healthy people (P＜0.05), but no significant difference was found between HD patients and PD patients. There were no significant differences of anti-erythropoietin antibody, Hb, BUN, Scr, iPTH and CRP among different primary diseases. Hb was negatively correlated with anti-erythropoietin antibody and CRP (r=-0.56, -0.20，P＜0.05), but was not correlated with BUN, Scr, iPTH. There was no correlation of anti-erythropoietin antibody with BUN, Scr, CRP and iPTH. One patient receiving recombinant human erythropoietin (rHuEPO) treatment with anti-erythropoietin antibody 43.63 U/L developed pure red cell aplasia diagnosed by marrow biopsy. Conclusions The anti-erythropoietin antibody levels of the dialysis patients are significantly higher as compared to healthy people, but are not significantly different between HD and PD patients. Anti-erythropoietin antibody is not correlated with BUN, Scr, iPTH and CRP. Hb is negatively correlated with anti-erythropoietin antibody and CRP. The rHuEPO can induce the anti-erythropoietin antibody leading to pure red cell aplasia in dialysis patients.

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Effects of urokinase-type plasminogen activator on high glucose-induced rat mesangial cells proliferation and phenotype transformation

CHENG Hui;DING Guo-hua;CHEN Cheng;SHI Ming;YANG Hong-xia.

2011, 27(7): 515-519.

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Objective To explore the effects and mechanisms of urokinase-type plasminogen activator (uPA) on high glucose-induced rat mesangial cells proliferation and phenotype transformation. Methods Rat mesangial cells were cultured and incubated in media containing either 5 mmol/L D-glucose or 30 mmol/L D-glucose with or without addition of wortmannin, or uPA (105 U/L) for different time periods. At the end of the incubation period, mesangial cells proliferation was assessed by MTT assay and flow cytometric analysis. Cyclin-dependent kinase 2 (CDK2) and p27kip1 expression and activation of Akt were evaluated by Western blotting and Akt kinase assay respectively. Furthermore, the expression and distribution of α-SMA were detected with laser confocal microscopy. Results MTT assay and flow cytometric analysis demonstrated that high glucose induced mesangial cells proliferation (P<0.05) and an incresed proportion of cells in G2/M+S stage after 24 h incubation (P<0.01), which were attenuated by uPA or wortmannin (P<0.01). High glucose induced the enhance of Akt activity after 3 h (P<0.05), and the effect was inhibited by wortmannin or uPA (P<0.01). High glucose did not alter CDK2 expression (P>0.05), but significantly inhibited p27kip1 expression (P<0.05), which was attenuated by wortmannin or uPA (P<0.01). High glucose induced the up-regulation of α-SMA expression and perinucleus location in mesangial cells after 24 h (P<0.01), which were alleviated by wortmannin or uPA (P<0.01). Conclusion uPA up-regulates p27Kip1 expression and counteracts high glucose-induced mesangial cells proliferation and phenotype transformation via blocking PI3K-Akt signaling pathway.

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Role of extracellular signal-regulated kinases in aldosterone-induced rat mesangial cells proliferation

YAO Li;SUN Li;WEI Min;GE Dan-mei;WANG Li-ning.

2011, 27(7): 520-524.

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Objective To determine the role of extracellular signal-regulated kinases (ERK1/2) in aldosterone-induced rat mesangial cells (RMCs) proliferation. Methods RMCs were obtained from intact glomeruli of 4- to 6-week-old Sprague-Dawley rats and characterized according to published methods. RMCs between passages 5 and passages 10 were used. Protein levels of mineralocorticoid receptor(MR) in RMCs were analyzed by Western blotting. The cells were divided into the following groups: control group, PD98059(10 ?滋mol/L) group, eplerenone (1 ?滋mol/L) group, aldosterone (100 nmol/L) group, aldosterone (100 nmol/L) ＋PD98059 (10 ?滋mol/L) group, aldosterone(100 nmol/L)＋eplerenone (1 ?滋mol/L) group. ERK1/2 activity was measured by Western blotting. Cell proliferation of RMCs was evaluated by [3H]-thymidine uptake measurements. Results MR protein expression in RMCs was confirmed by Western blotting. Aldosterone activated ERK1/2, and the maximal ERK1/2 activation induced by aldosterone was at a concentration of 100 nmol/L. Aldosterone (100 nmol/L)-induced activation of ERK1/2 peaked at 10 minutes (P<0.05). Pretreatment with a selective MR antagonist eplerenone (1 ?滋mol/L) significantly attenuated aldosterone-induced ERK1/2 phosphorylation. Aldosterone (100 nmol/L) treatment for 30 hours increased [3H]-thymidine incorporation of RMCs (135%±8% of controls, P<0.05). Cellular proliferation induced by aldosterone could be prevented by pretreatment with eplerenone or an ERK (MEK) inhibitor PD988059. Conclusion Aldosterone induces RMCs proliferation through MR and ERK1/2 activation, which may contribute to the pathogenesis of glomerular mesangial injury.

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Comparison of kidney tubular epithelial cells and endothelial cells grown on titania nanotubes

LIU Hui-qiong;ZHU Wen;LIU Jian-feng;LIU Xi;TONG Da-li.

2011, 27(7): 525-529.

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Objective To observe the adhesion and growth of LLC-PK1 cells and ECV304 cells on titania nanotube arrays, and provide evidence for construction of miniaturation bioartificial kidney. Methods Four different diameters nanotube materials were prepared by anodic oxidation, each material was processed by unannealed and with UV irradiation, annealed and without UV irradiation, annealed and with UV irradiation, respectively, which had 12 groups totally, then two kinds of cells were separately grown on the 12 materials. The adhesion and growth of the two kinds of cells were studied under a fluorescence microscope. MTT assay was used to test the activity of two kinds of cells on different diameters and the proliferation of two kinds of cells on 70 nm diameters. Results The adhesion and proliferation of two kinds of cells on TiO2 nanotube arrays were basically consistent, both on anatase TiO2 nanotubes with 70 nm diameter but without UV irradiation showed the optimal adhesion and activity. The activities of LLC-PK1 cells and ECV304 cells were both increased with time extended, while the absorbance of ECV304 cells was higher on pure Ti film than on titania nanotube. Conclusion TiO2 nanotube is beneficial to LLC-PK1 cells, but is unfavorable for ECV304 cells when they grow alone.

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IgA肾病专题

临床研究

基础研究

IgA肾病专题

临床研究

基础研究