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Objective:To investigate the urinary sediment findings and the clinicopathologic features of IgA nephropathy (IgAN) patients with acute kidney injury (AKI).Methods:It was a retrospective study. The patients with renal biopsy-proven primary IgAN in Peking University First Hospital from January 31, 2013 to July 31, 2015 were selected. According to whether AKI occurred at renal biopsy or not, the patients were divided into AKI group and non-AKI group. Morning urine samples were obtained on the day of renal biopsy. Urine sediments, including various cells and casts, were examined. The clinical data, urinary sediments, and renal pathological changes were compared between the two groups. Logistic regression analysis was performed to identify the association between clinical pathological changes, urinary sediment indicators and AKI, or clinical pathological changes and urinary sediment indicators.Results:There were 502 IgAN patients enrolled in this study, with age of (36.1±12.1) years old and 261 males (52.0%). The incidence of AKI was 11.4% (57/502) among the enrolled patients at the time of renal biopsy. Common causes of AKI included gross hematuria-induced AKI (10 cases), acute tubulointerstitial nephritis (10 cases), crescentic IgAN (9 cases), malignant hypertensive renal damage (6 cases), and multiple etioloqy or unknown etiology (22 cases). Compared with non-AKI group, AKI group had higher proportions of males and malignant hypertension, higher levels of proteinuria and urinary erythrocyte counts, and higher frequencies of gross hematuria, leukocyturia, renal tubular epithelial cells, and granular casts (all P<0.05). AKI group also had higher proportions of severe tubular atrophy/interstitial fibrosis (T2) and cellular/cellular fibrous crescent formation (C2) than non-AKI group (both P<0.05). Logistic regression analysis results showed that, there were statistically significant differences in the correlation between AKI and gender, 24 h urinary protein, urinary erythrocyte counts, granular casts and renal tubular atrophy/interstitial fibrosis (T) scores (all P<0.05). Hematuria, leukocyturia, red blood cell casts, white blood cell casts, granular casts, and fatty casts were correlated with endothelial hypercellularity (E) and cellular/cellular fibrous crescent formation (C) scores, respectively (all P<0.05). Hematuria was correlated with mesangial hypercellularity (M) scores ( OR=2.613, 95% CI 1.520-4.493, P=0.001). Hematuria ( OR=1.723, 95% CI 1.017-2.919, P=0.043) and fatty casts ( OR=2.646, 95% CI 1.122-6.238, P=0.026) were correlated with segmental sclerosis or adhesion (S) scores. Leukocyturia ( OR=1.645, 95% CI 1.154-2.347, P=0.006) and fatty casts ( OR=2.344, 95% CI 1.202-4.572, P=0.012) were correlated with T scores. Epithelial cell cast was correlated with C scores ( OR=1.857, 95% CI 1.174-2.939, P=0.008). Conclusions:AKI is a common complication among IgAN patients with diverse etiology and more severe clinicopathological features. Urinary sediment findings can reflect renal pathological changes to some extent, and therefore assist in the clinical diagnosis and treatment of IgAN patients with AKI.
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Automated urine analysis poses the advantage of rapid turn around time and is suitable for initial screening of large number of samples.However,standardized manual microscopic examination of urine sediment remains valuable for a variety of differential diagnosis,such as screening the potential causes of hematuria,to predict proliferative and non-proliferative renal pathological injury. Urinary sediment scoring system is often used to differentiate acute tubular necrosis (ATN) from pre-renal acute kidney injury (AKI).The urinary podocytes serves as a marker of glomerular injury to determine the location of and to monitor activity of glomerular lesions,as well as to differentiate focal segmental glomerulosclerosis (FSGS) and minimal change disease (MCD).These useful analyses shows the importance of urine microscopy in the clinical practice of nephrology.( Chin J Lab Med,2012,35:780-783)
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Objective: To investigate whether hypoxia can affect the expression and secretion of connective tissue growth factor(CTGF) and fibronectin(FN) in primary cultured rat renal cortical myofibroblasts . Methods: The primary cultured rat renal cortical myofibroblasts were subjected to hypoxic (1%O_2) or normoxic (21% O_2) conditions for a variety of times. The protein levels of HIF - 1?, CTGF and FN protein were analyzed by Western blotting in both the whole cell lysates and supernatant culture medium 6 h,12 h and 24 h after incubation, respectively. RT-PCR was carried out to measure the levels of FN mRNA at different time points (2 h,3 h,6 h and 12 h). The activity of gelatinase MMP-2 and MMP-9 in the supernatant from the cultured cell medium was assayed by gelatin zymography. Results: The expression of HIF - 1?was induced at h6 in cells under hypoxia incubation. The levels of cellular CTGF protein were increased in hypoxia treated myofibroblasts at h6 (175%?52%),significantly elevated at h12 (347%?67%,P
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Objective:To observe the expression of connective tissue growth factor(CTGF) and its receptor-low density lipoprotein receptor-related protein (LRP), and the relevant signaling pathway for the regulation by long-term high glucose exposure in cultured podocytes. Methods:The effects of high glucose on the expression of CTGF and its receptor LRP were analyzed by western blotting. The activation of mitogen activated protein kinase ( MAPKS )signaling pathway by high glucose was also examined. Results: Basal levels of CTGF were observed in cultured mouse podocytes, the levels of CTGF protein were increased by high glucose medium groups on the 2nd day, reached the peak on the 4th day(P0.05).The levels of CTGF expression in normal glucose and mannitol glucose groups did not change markly. High glucose medium induced phosphorylation of ERK_ 1/2 at as early as minute 30, reached the peak at hour 6; maintained the activity at hours 12 and 24, and declined to the basal level at hour 48. However, phosphorylation of ERK_ 1/2 was not detected in normal glucose and mannitol glucose groups. Blockade of phosphorylation of ERK_ 1/2 with PD98059, a specific ERK_ 1/2 activation inhibitior, did decrease the high glucose-triggered expression of CTGF protein in 4 days. High glucose had no effect on the expression of LRP protein at each time point. Conclusion: Acute high glucose (2-4 days)stimulated the expression of CTGF protein via ERK_ 1/2-dependent signaling pathway in cultured podocytes, while cultured in high glucose for 6-8 days, the podocytes did not increase its CTGF level. Long-term high glucose had no effect on the expression of LRP in podocytes.
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AIM A&A treated PAN rats, and to observe the effects of A&A on MAPK signaling pathway. METHODS Rats were divided into control, PAN, A&A treated PAN (A&A) and enapril treated PAN (ACEI) groups. The pathological lesion was observed under a light microscope. Immunohistochemistry combined with semi quantitive method was used to investigate the following parameters: cell number, ? SMA expression and extracellular matrix deposition. Expression and phosphorylation of protein kinases ERK, JNK and p38 were assayed. RESULTS In PAN rats, A&A suppressed ? SMA expression, which was closely correlated to cell proliferation, and extracellular matrix accumulation in glomerular mesangium. A&A significantly attenuated ? SMA expression in the tubulo interstitial area which was also parallel to the renal interstitial fibrosis.In this study, expression of all subtypes of MAPK had no difference between control and PAN groups. Compared with the inactivation of ERK and p38, phosphorylation of JNK was observed in glomeruli, renal tubules and interstitial cells in PAN rats, which was also inhibited by A&A treatment. CONCLUSION The inhibitory effect of A&A on phenotypic changes of renal resident cells, especially glomerular mesangial cell, may participate in its renal protective mechanisms. This effect, at least partially, was mediated by down regulated JNK activation.
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Objective: To investigate peroxisome proliferator activated receptor ? (PPAR ?) expressions in patients with actively proliferating glomerulonephritis such as type Ⅳ lupus nephritis (LN) and cellular crescentic glomerulonephritis (RPGN). Methods: All patients were divided into 4 groups as follows: RPGN (17 cases); LN type Ⅳ (15 cases); mild mesangial proliferative IgA nephropathy (IgAN, 7 cases) and minimal change disease (MCD, 10 cases). Clinical parameters, immunohisto chemistry stain and in situ hybridization of renal biopsy specimens were performed. Results: Clinically, proteinuria and hematouria were increased and Ccr were decreased in LN and RPGN patients, and increased ESR and decreased complement C3 were found in group LN. Active index of renal specimens were significantly higher in LN and RPGN groups than in IgAN and MCD groups. Renal specimens of MCD patients showed no positive PPAR ? staining in all sections; little immunoreactivity was detected in sections of glomerular, tubular and interstitial cells from IgAN patients. Glomerular positive staining of PPAR ? in renal sections in LN and RPGN patients[(3.3?1.8) and (2.8?1.2) cells per section of glomerulus, respectively] were significantly increased compared with that in IgAN patients [(0.7?0.5) cells per section of glomerulus]. Similarly, tubular positive staining of PPAR ? in LN and RPGN patients (27.38? 12.46, 23.36?10.55) were also elevated compared with that in IgAN and MCD patients(6.51?3.49, 1.72?0.31). The relevance assay results showed positive relationship between active index and glomerular or tubular PPAR ? immunohisto chemistry staining cell numbers (0.478, P
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Objective:To investigate the anti-fibrotic effect of sirolimus(rapamycin)at the cell level.Methods:The primary cultured rat renal cortical myofibroblasts were divided into two groups,control group and sirolimus 40 mg/L group at each time point.The protein levels of ?-SMA,Col-Ⅰ,fibronectin(FN)were analyzed by Western blot in both the whole cell lysates and supernatant culture media 12 h,24 h and 48 h after incubation,respectively.Real-time quantitative PCR was carried out to measure the levels of procollagen-ⅠmRNA 1 h,2 h,4 h,and 6 h after cell incubation.The activities of gelatinase MMP-2 and MMP-9 in the supernatant from the cultured cell media were assayed by gelatin zymography.Results:(1)Sirolimus had no effect on the expression of ?-SMA of myofibroblasts at differnet time points.(2)The expression of Col-Ⅰin the whole cell lysates both reduced at the end of 24 h and 48 h in sirolimus group significantly [(0.58?0.05)and(0.63?0.18),P
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<p><b>OBJECTIVE</b>To identify genes associated with the chronic progression of renal disease and a stragalus and angelica (A&A)'s renal protective effects.</p><p><b>METHODS</b>The technique of silver staining mRNA differential display (DD) was used to investigate changes of gene expression in normal, sclerotic and A&A treated sclerotic kidneys. We isolated genes differentially expressed during the progression of renal disease which could be normalized by A&A.</p><p><b>RESULTS</b>Several genes related to A&A's protective effects were isolated and one of them was confirmed by Northern blot.</p><p><b>CONCLUSION</b>Silver staining mRNA differential display is a simple and effective technique for isolating differentially expressed genes. The isolated new gene may be related to the progression of chronic renal disease and contribute to A&A's protective effects.</p>
Subject(s)
Animals , Male , Rats , Angelica sinensis , Astragalus Plant , Drugs, Chinese Herbal , Pharmacology , Kidney , Metabolism , Kidney Diseases , Drug Therapy , Metabolism , Phytotherapy , Polymerase Chain Reaction , Protective Agents , Pharmacology , RNA, Messenger , Rats, Sprague-Dawley , Staining and LabelingABSTRACT
AIM: To investigate the effect of hypoxia on the myofibroblast transdifferentiation from fibroblasts,and associated signaling of hypoxia on the production of collagen Ⅰ in cultured rat renal cortical myofibroblasts.METHODS: The study is composed of two relevant parts.In the first part,a normal rat renal interstitial fibroblast cell line NRK-49F was treated with hypoxia(1% O2) or normoxia(21% O2) for 6 h,12 h and 24 h.The expression of hypoxia inducible factor-1?(HIF-1?) was examined by Western blotting in order to make sure the hypoxic condition is reliable.The myofibroblast transformation from fibroblasts induced by hypoxia was assayed by detecting the protein levels of ?-smooth muscle actin(?-SMA).In the second part,the object was done on the primary cultured rat renal cortical myofibroblasts.Myofibroblasts were subjected to hypoxic or normoxic conditions for variety of times.The levels of HIF-1? in cell lysates and collagen I protein in supernatant culture medium and the activation of extracellular signal-regulated kinase(ERK)1/2 MAPK pathway were analyzed by Western blotting.RT-PCR was carried out to measure the levels of collagen I mRNA at different time points(2 h,4 h and 6 h).The distribution of HIF-1? in myofibroblasts was demonstrated by immunocytochemistry.The changes of collagen I production were detected after PD98059,a specific inhibitor of ERK1/2 activation pretreatment and during the hypoxia incubation.The activity of gelatinase matrix metalloproteinase-2(MMP-2) and MMP-9 in the supernatant medium from the cultured cells were assayed by gelatin zymography.RESULTS: Significant increased levels of HIF-1? protein appeared in cell lysates under hypoxia for 6 h.Furthermore,HIF-1? was translocated into nuclei of myofibroblasts after 6 h exposure of myofibroblasts to hypoxia.The levels of ?-SMA protein increased in NRK-49F under hypoxia for 12 h(187%?32%,P
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Macrophages have been identified to play an important role in acute and chronic renal injury through exerting multiple biological effects.This review focus on the origin of macrophages in kidney, mechanism of renal injury and strategy of inhibiting macrophage infiltration. [
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AIM: To understand the expression of low density lipoprotein receptor-related protein (LRP) in tubulointerstitial fibrosis of unilateral ureter obstruction (UUO) rats. METHODS: The localization of LRP within kidneys were assessed by immunohistochemical staining, the protein level of LRP and connective tissue growth factor (CTGF) in kidney were analysised by Western blot. RESULTS: The location of LRP positive-staining and the protein level of LRP were in the same tendency with CTGF, the level of LRP had strong correlationship with the level of CTGF (r=0.786, P
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AIM: To compare the renoprotective effects of angiotensinⅡtypeⅠreceptor antagonist (AT 1RA) with that of angiotensin converting enzyme inhibitor (ACEI) and to investigate their influences on intrarenal renin-angiotensin system. METHODS: Experimental nephrotic syndrome model was induced in SD rats with repeated peritoneal injections of puromycin. Twenty-eight rats were randomly divided into four groups: normal control, nephrotic control, ACEI-treated and AT 1RA-treated group. Serum, urine and renal tissue were collected for study 12 weeks later. RESULTS: The urine protein was less and renal function was better in both treated groups. The glomerular and interstitial injury indexes of both ACEI and AT 1RA treated rats were lower than that of nephrotic control rats and had no significant difference between the two treated groups. The renal local ACE activity and angiotensinⅡ of nephrotic control group were significantly higher than that of normal control group and the two treated group(P
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Objective To investigate the feasibility whether clearance of iohexol can be a reliable, sensitive and safe method for the determination of GFR. Methods The GFR of 19 patients with different renal functions were examined using clearance of 99m Tc-DTPA and clearance of iohexol. Then the correlation of them was analyzed. Serum and urinary iohexol was determined by X-ray fluorescence analysis. Results, These two methods were significantly correlated (r = 0. 98). Conclusion Clearance of iohexol is a safe, comfident, no-radioactivity method for the clinical practice of GFR determination.
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AIM: To investigates the role of phosphatidic acid (PA) in the expression of several inflammatory mediators produced by glomerural macrophages (GM?). METHODS: The study was performed on a rat model of accelerated anti-glomerural basement membrane (anti-GBM) glomerulonephritis (GN). GN-GM? were isolated and identified. Peritoneal M? (P-M?) of both normal and GN rats were used as controls. Block and reverse test were investigated with rhIL-1? stimulated, lisofylline (LSF) and phosphatidic acid (PA). Macrophage expression of ICAM-1 and TGF-? 1 were assessed at the level of protein and gene by immunocytochemistry, northern blot and RT-PCR. RESULTS: (1) After stimulated with rhIL-1?, GN-GM? produced much more ICAM-1, MCP-1 and TGF-? 1 than P-M?, and it's gene expression was similar as protein product. (2) mRNA expression of these factors was up-regulated again after the GN-GM? were pretreated with LSF then PA was added. CONCLUSIONS: Since GN-GM? plays an important role for PA in the mediation of glomerular injury, inhibiting of PA production is the keypoint of blocking M? mediated inflammatory effects. LSF may be an effective medicine in therapy for acute inflammatory forms of GN.
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AIM: To study the expression of connective tissue growth factor (CTGF) induced by hypoxia, and the role and mechanism of hypoxia on promoting renal interstitial fibrosis. METHODS: Renal interstitial fibrosis was induced by unilateral ureteral obstruction (UUO) in rat animal model for 9 days as in vivo studies; marker of hypoxia-HIF-1? mRNA and protein, the expression of CTGF in the obstructed kidneys were assessed by RT-PCR, immunohistochemistry and Western blotting respectively. In vitro , normal rat kidney interstitial fibroblast cells (NRK-49F) were exposed to hypoxia (1%O 2) for up to 6 hours, hypoxia was confirmed by detecting the expression of HIF-1? protein in cells, cellular level of CTGF mRNA and protein were assessed by RT-PCR and Western blotting respectively. RESULTS: Neither HIF-1? mRNA nor HIF-1? protein was expressed in the kidney from sham-operated group of rats. High level of HIF-1? mRNA were occurred, and strongly HIF-1? positive immunostaining were seen in the tubular and interstitial cells in kidney from UUO rats. Expression and location of CTGF protein were paralleled and relevant with the expression of HIF-1? protein in kidney of UUO rats. In cultured NRK-49F cell line, subjected to hypoxia even for 6 hours stimulated the expression of CTGF mRNA and protein. CONCLUSION: Our results indicated that hypoxia could stimulate the expression of CTGF mRNA and protein in kidney from UUO rats, which may in turn contribute to renal interstitial fibrosis.