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La nefropatía por inmunoglobulina M (NIgM) es una glomerulopatía idiopática caracterizada por depósitos mesangiales globales y difusos de IgM. Se realizó un estudio retrospectivo de las características clínicas e histopatológicas de los pacientes con NIgM atendidos en nuestro servicio. De 241 biopsias renales, 21 correspondieron a NIgM (8,7 %). Se incluyeron 18 pacientes (14 de sexo femenino, mediana de edad: 3,08 años). Se excluyó a 1 paciente por enfermedad sistémica asociada y a 2 por seguimiento menor a 1 año. Catorce pacientes se manifestaron con síndrome nefrótico (SN) y 4 con proteinuria aislada o asociada a hematuria. En la microscopia óptica, 13 presentaron hiperplasia mesangial, y 5 esclerosis focal y segmentaria. De los pacientes con SN, 7 fueron corticorresistentes, 4 corticodependientes y 3 presentaban recaídas frecuentes. Todos los pacientes con SN y 1 con proteinuria-hematuria recibieron inmunosupresores; los 18 pacientes recibieron, además, antiproteinúricos. Luego de 5,2 años (2-17,5) de seguimiento, 6 pacientes evolucionaron a enfermedad renal crónica
Immunoglobulin M nephropathy (IgMN) is an idiopathic glomerulopathy characterized by diffuse global mesangial deposits of IgM. We retrospectively studied the clinical and histopathological characteristics of the patients with IgMN seen in our service. Of 241 renal biopsies, 21 corresponded to IgMN (8.7 %). One patient was excluded due to associated systemic disease and 2 due to follow-up less than 1 year, 18 were included (14 girls, median age 3.08 years). Fourteen manifested with nephrotic syndrome (NS) and the remaining with proteinuria (isolated or associated with hematuria). On light microscopy, 13 had hyperplasia with mesangial expansion and 5 had focal and segmental sclerosis. Of the patients with NS, 7 were steroid-resistant, 4 steroid-dependent, and 3 frequent relapsers. All patients with NS and 1 with proteinuria-hematuria received immunosuppressants; the 18 patients also received antiproteinuric drugs. After 5.2 years (2-17.5) of follow-up, 6 patients developed chronic kidney disease.
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Humans , Male , Female , Infant, Newborn , Infant , Child, Preschool , Child , Adolescent , Immunoglobulin M , Nephrotic Syndrome/pathology , Nephrotic Syndrome/therapy , Kidney Diseases , Nephrotic Syndrome/diagnosisABSTRACT
Objective To investigate the effect of tumor necrosis factor-α (TNF-α) on intracellular calcium concentration ([Ca2 +] i) and the contraction of glomerular mesangial cells (GMCs),and prove that hypercontractility of GMCs induced by TNF-α in hepatorenal syndrome(HRS) was connected with inositol 1,4,5-trisphophate receptors (IP3Rs).Methods GMCs were divided into TNF-α-treated 0 h,4h,and 24 h groups.Another 3 groups were blocked by 2-APB.The effect of TNF-α on [Ca2 +] i was identified and observed whether it could be blocked by 2-APB.Contraction of GMCs was determined by accessing the surface area of cells before and after contraction.Results TNF-α significantly increased ET-induced calcium release,in that we found higher [Ca2 +] i after stimulated by ET in TNF-α-treated 4 h group and 24 h group[4 h:(648.08 ±267.11) nmol/L; 24 h:(879.30 ±-260.29) nmol/L; 0 h:(619.93 ±258.94)nmol/L,F =5.486,4 h vs 0 h:P ＜ 0.05 ; 24 h vs 0 h:P ＜ 0.05 ;24 h vs 4 h:P ＞ 0.05].This phenomenon can be totally blocked by 2-APB in all groups.The change in planar surface area in response to ET was slightly in control cells but significantly enhanced in TNF-α-treated cells [4 h:(2198 ± 340)μm2; 24h:(2260±553)μm2; 0 h:(2436±474)μm2,F =4.001,4 h vs0 h:P ＜0.05; 24 h vs0 h:P ＜0.05;24 h vs 4 h:P ＞ 0.05].Conclusion TNF-α can enhance ET-induced sarcoplasmic reticulum Ca2 + release and increase the contractile responses of GMCs to ET,which is associated with IP3Rs.TNF-α is responsible for hyperconstractility of glomeruli in HRS.
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Objective To explore the effect of angiotensin-(1-7) [Ang-(1-7)] on proliferation and secretion in cultured rat's glo-merular mesangial cells(GMC) induced by transforming growth factor-β1 (TGF-β1). Methods GMC in logarithmic growth phase were in-cubated, and then were divided into 4 groups: control, Ang-(1-7), TGF-β1,and TGF-β1 + Ang-(1-7) group. Cell numbers of rat's GMC were detected by WST-1. Laminin(LN) and collagenlV (ColⅣ) mRNA expressions in GMC were analyzed by RT-PCR. The secretion of LN and ColⅣ in culture medium of rat's GMC was measured by radioimmunoassay. Results Ang-(1-7) significantly inhibited basal and TGF-β1-induced proliferation of cultured glomerular mesangial cells. Ang-(1-7) significantly down-regulated LN and Col Ⅳ mRNA expressions in glomerular mesangial cells and also inhibited the secretion of LN and ColⅣ induced by TGF-β1(P <0. 05). Conclusions Ang-(1-7) can inhibit basal and TGF-β1-induced proliferation, and inhibit ECM secretion of cultured rats glomerular mesangial cells.
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Objective To study the effect of high glucose and lysophosphatidylcholine (LPC) on the role of fibronectin and plate-let activating factor (PAF) via the interaction of endothelial cells and mesangial cells. Methods The model of intercellular interaction be-tween endothelial cells and mesangial cells was established and divided into 4 groups: control, mannitol, high glucose and LPC, and BN52021 group. The level of fibronectin and PAF were determined by enzyme linked immunosorbent assay in the culture media. Results The level of fibronectin and PAF of high glucose and LPC group were higher than those of control group in co-culture and monolayer cell cul-ture (P<0.05). Intervened by high glucose and LPC, the level of fibronectin and PAF of co-culture were higher than those from monolay-er cell culture (P<0.05). The level of fibrocentin in the BN52021 group was lower than that of high glucose and LPC (P<0.05). Con-clusions Exposed to high glucose and LPC, endothelial cells and mesangial cells can interact with each other to produce more fibrocentin and PAF. The increase of fibronectin is partly concerned with PAF.
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promoted by high glucose, which can enlarge the biological effect of PAF.
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Objective To study the effects of keto-valine-calcium and keto-isoleucine-calcium on(human mesangial cells)HMCs. Method HMCs were stimulated by keto-valine-calcium and keto-isoleucine-calcium. The AT1R and TGF-β1 were detected. MTr method was used to measure the proliferation of HMCs, and cell cycle was studied by flow cytometry. Results The expression of AT1R and TGF-β1was increased in the experiment groups compared with negative and DMSO control groups. Cell cycle G1 attest and cell apoptosis were observed in the experiment groups. Conclusions 10mM keto-valine-calcium and keto-isoleueine-calcium have multiple effects on HMCs in vitro, which not only increased the expression of AT1R,but also the expression of TGF-β1.Furthermore,keto-valine-calcium and keto-isoleucine-calcium can induce cell cycle G1 arrest and cell apoptosis.
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Objective To investigate the effects of Lp(a)on proliferation GMCs of rat model induced by lipopolysaccharide and explore the possible mechanism of Lp(a)in the proliferation of rat GMCs.Methods To observe the effects of Lp(a)on proliferation of GMCs,different dosage of Lp(a)were used,The research were divided into three groups:Control group,LPS group,Lp(a)group.After culture(at the end of 12h,24h,48h,60h and 72h),the cultured GMCs and suspension were collected to observe the rate of GMCs proliferation by MTT,the positive rate of proliferation cell nuclear antigen(PCNA)by immunohistochemisty,and the level of intercellular adhesion molecule-1(ICAM-1)by ELISA respectively.Results Compared with control and LPS group,MTT,positive rate of PCNA and ICAM-1 of GMCs were increased more significantly in Lp(a)group.MTT ,the positive rate of PCNA and ICAM-1 of GMCs were increased as Lp(a)dosage increased,a maximal effect was seen when Lp(a)was 2.5 μg/L or 5.0μg/L.When the dosage continue increased,MTT,the positive rate of PCNA and ICAM-1 activity of GMCs began to decrease in Lp(a)group.ICAM-1 showed positive correlation with MTT and the positive rate of PCNA.Conclusion Lp(a)can significantly affect the rate of GMCs proliferation,and this affection is in a dosage-and timedependent manner.Low dosage stimulates GMCs proliferation, and high dosage inhibits GMCs proliferation.ICAM-1 shows positive correlation with MTT and the positive rate of PCNA.The effect of Lp(a)on GMCs may be through ICAM-1.
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The extracellular calcium sensing receptor (CaSR) belongs to the type III family of G-protein-coupled receptors, a family that comprises the metabotropic glutamate receptor and the putative vomeronasal organ receptors. The CaSR plays an important role for calcium homeostasis in parathyroid cells, kidney cells and other cells to directly 'sense' changes in the extracellular calcium ion concentration ((Ca2+)o). The mesangial cells are known to be involved in many pathologic sequences through the mediation of altered glomerular hemodynamics, cell proliferation, and matrix production. In this study, we examined the expression of the CaSR in the mouse mesangial cell lines (MMC, ATCC number CRL-1927). Reverse transcription- polymerase chain reaction (RT-PCR) was perform with CaSR-specific primers, and this was followed by nucleotide sequencing of the amplified product; this process identified the CaSR transcript in the MMCs. Moreover, CaSR protein was present in the MMCs as assessed by Western blot and immunocytochemical analysis using a polyclonal antibody specific for the CaSR. Functionally, (Ca2+)o induced the increment of the intracellular calcium concentration ((Ca2+)i) in a dose-dependent manner. This (Ca2+)i increment by (Ca2+)o was attenuated by the pretreatment with a phospholipase C inhibitor (U73122) and also by a pretreatment with a CaSR antagonist (NPS 2390). The similar results were also obtained in IP3 accumulation by (Ca2+)o. To investigate the physiological effect of the CaSR, the effect of the (Ca2+)o on cell proliferation was studied. The increased (Ca2+)o (up to 10 mM) produced a significant increase in the cell numbers. This mitogenic effect of (Ca2+)o was inhibited by the co-treatment with a CaSR antagonist. From these results, the (Ca2+)o-induced (Ca2+)i elevation in the MMC is coupled with the extracellular calcium sensing receptor. Furthermore, (Ca2+)o produces a mitogenic effect in MMCs.
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Animals , Mice , Calcium/metabolism , Cell Line , Cell Proliferation , Inositol 1,4,5-Trisphosphate/metabolism , Mesangial Cells/cytology , RNA, Messenger/genetics , Receptors, Calcium-Sensing/geneticsABSTRACT
Objective: It was reported that major subclasses of anti-golmerular basement membrane (GBM) antibody were IgG1 and IgG4. The IgG1 subclass was mainly found in male patients and IgG4 subclass in female patients with anti-GBM disease. This study investigates the distribution of anti-GBM IgG subclasses and their association with clinical characteristics. Methods: Sera from 50 patients diagnosed as anti-GBM disease during 1991-2003 were collected. The anti-GBM antibodies with IgG1, IgG2, IgG3, and IgG4 subclasses were detected with purified bovine ?(IV)NC1 as solid phase ligand in ELISA. The association with clinical manifestations was further investigated. Results: The positive rates of IgG anti-GBM subclasses were IgG1 94%, IgG2 56%, IgG3 12%, IgG4 88%, respectively. IgG1 and IgG4 were the main subclasses in anti-GBM IgG subclasses (P
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Objeetive To explore the relationship among the glomerular mesangium components,degree of renal lesion and glomerulosclerosis in patients with type 2 DM.Method A quantitative study of type Ⅳ collagen in kidneys taken from twenty-one patients with type 2 DM and ten-two control autoptical renal tissue specimens were investigated by immunochemical techniques,computer image analysis system.Results The amounts of type Ⅳ collagen were increased parallel with advances of glomerulosclerosis and renal arteriolosclerosis in patients with type 2 DM,but the collagen had not obviously elevation in renal failure patients.Conclusions The increased type Ⅳ collagen is a characteristic manifestation of early and moderate lesions of glomerulosclerosis in type 2 DM.Quantitative determination of glomerular mesangium components could be a good index to judge the lesion degree of kidney.
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AIM: To explore the mechanism of the nitric oxide (NO) synthesis induced by human C5b-9 complex in glomerular mesangial cells(MC) of rats. METHODS: The MC of rats were cultured and stimulated with human complement C5b-9 complex to induce TNF? and IL-1?. At the same time, several parameters related to NO synthesis were measured at 3 h, 6 h and 24 h after C5b-9 stimulation. The effects of monoclonal antibodies against TNF? and IL-1? on NO synthesis were examined in this system. RESULTS: TNF? concentration in supernatant from MC in C5b-9 group was higher than that of control group at 6, 24 h after stimulation with C5b-9 complex and reversed by adding anti-TNF? McAb. C5b-9 complex didn't stimulate the release of IL-1? in same system. In addition, the expression of iNOS mRNA in MC was observed at 3 h after stimulation with C5b-9. Levels of iNOS mRNA expression and cGMP in MC and NO - 3/NO - 2 in supernatant from MC in C5b-9 group were higher than those in control group at 6, 24 h after C5b-9 stimulation, these changes were also reversed by adding monoclonal antibody against TNF?. CONCLUSION: C5b-9 complex could induce iNOS mRNA expression at 3 h after C5b-9 stimulation, and the synthesis of NO at 6, 24 h was related to TNF? released from cultured MC of rats by C5b-9 complex to a certain extent.
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AIM: To investigate the effects of advanced glycation end products (AGEs) on the expression of plasminogen activator inhibitor-1 (PAI-1) in rat mesangial cells and its relationship with extracellular matrix accumulation. METHODS: Rat mesangial cells were treated with AGE-modified bovine serum albumin or native bovine serum albumin. Normal mesangial cells without any treatments were used as control. Fibronectin (FN), collagen Ⅳ, PAI-1 protein contents were detected by ELISA. PAI-1 mRNA was analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: AGEs (0-200 mg/L) did not influence mesangial cells proliferation, but stimulated FN , collagen IV and PAI-1 contents in mesangial cell cultured medium in different degrees. AGEs also increased PAI-1 mRNA expression. CONCLUSION: AGEs increase the expression of PAI-1 in rat mesangial cells. AGEs may reduce ECM degradation through increasing PAI-1 expression, which may be one of the mechanisms of ECM accumulation in diabetic nephropathy.
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AIM: To investigate the effects of Lipopolysaccharide(LPS) and interieukin 1 receptor antagonist (IL- 1ra) on mesangial cells proliferation and nitric oxide synthesis. METHODS: Glomerular mesangial cells from SD rats were cultured. The first and second passages of cultured cells were used for the experiment. LPS and LPS plus IL- 1ra were added in cell cultures, respectively. By using chemical method the nitrite in supernatants was measured ,3H- TaR incorporation was determined to evaluate the GMC proliferation. Northern and slot hybridizations were performed to detect the expression of iNOS mRNA. RESULTS: There were expression of iNOS mRNA, more production of nitrite(0.64 + 0.25 vs 0. 12 + 0.06 nmol/104 cell) in supernatants and GMC proliferation(3735 + 1177.9 vs 1785 + 280.6) in LPS group compared to the control. While compared with LPS group, in LPS + IL- 1ra GMC group, expression of iNOS mRNA decreased by 40%, nitrite increased(3.28 + 0.33 nmol/104 cell), proliferation of GMC decreased (818 + 77.27). CONCLUSION: LPS could activate the GMC to express iNOS mRNA and produce more nitrite. IL - 1ra could partially inhibit the effects of LPS on the expression of iNOS mRNA in GMC, but not nitrite. There is no synchronous correlation between NO production and GMC proliferation.
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AIM: To investigate the effects of Lipopolysaccharide(LPS) and interleukin 1 receptor antagonist(IL-1ra) on mesangial cells proliferation and nitric oxide synthesis. METHODS: Glomerular mesangial cells from SD rats were cultured. The first and second passages of cultured cells were used for the experiment. LPS and LPS plus IL-1ra were added in cell cultures, respectively. By using chemical method the nitrite in supernatants was measured,3H-TdR incorporation was determined to evaluate the GMC proliferation. Northern and slot hybridizations were performed to detect the expression of iNOS mRNA. RESULTS: There were expression of iNOS mRNA, more production of nitrite(0.64?0.25 vs 0.12?0.06 nmol/104 cell) in supernatants and GMC proliferation(3735?1177.9 vs 1785?280.6) in LPS group compared to the control. While compared with LPS group, in LPS+IL-1ra GMC group, expression of iNOS mRNA decreased by 40%, nitrite increased(3.28?0.33 nmol/104 cell), proliferation of GMC decreased (818?77.27). CONCLUSION: LPS could activate the GMC to express iNOS mRNA and produce more nitrite. IL-1ra could partially inhibit the effects of LPS on the expression of iNOS mRNA in GMC, but not nitrite. There is no synchronous correlation between NO production and GMC proliferation.
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Objective To investigate the role of hexosamine biosynthesis pathway (HBP) in the functionalalterationofmesangialcellstransinfectedwith glucose transporter 1 (GLUT1) gene. Methods Rat mesangial cells were transinfected with the human GLUT1 gene (MCGT1) by retrovirus vector. Mesangial cells transinfected with bacterial ?-galactosidase (MCLacZ) were used as control. Glucose uptake was detected with 2-deoxy-〔 3H〕-D-glucose (2-DG). Cell size, RNA/DNA ratio, protein/DNA ratio and the synthesis of fibronectin were evaluated by flow cytometry. The activity of glutamine: fructose-6-P aminotransferase (GFAT), which is the key enzyme of HBP, was assayed by spectrophotometry. The expression of GFAT gene was analyzed by RT-PCR. Results MCGT1 demonstrated higher 2-DG uptake than MCLacZ 〔(741.0?60.5)dpm/?g protein vs (92.2?9.0)dpm/?g protein,P
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AIM: To investigate the effect of L-arginine(L-arg) on the production of collagens of human mesangial cells. METHODS: Radioimmunoassay, hydroxyproline colorimetric assay and reverse transcription polymerase chain reaction (RT-PCR) were used to determine procollagen Ⅲ, total collagen level in the supernatant and expression of collagen Ⅳ mRNA in human mesangial cells. RESULTS: L-arg significantly inhibited the production of procollagenⅢ, total collagen in the supernatants ( P
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AIM: To investigate the effects of interleukin-13(IL-13) on the inflammatory reaction of glomerular mesangial cells. METHODS: TNF-? protein in the supernatant of cultured mesangial cells was determined with ELISA. ICAM-1 expression on the mesangial cells was determined with flow cytometry. The expression of TNF-? mRNA and ICAM-1 mRNA were semiquantatively tested with RT-PCR. RESULTS: Mesangial cells did not constitutively express TNF-? mRNA and TNF-? protein. Induced by LPS(10 mg/L) for 24 h, mesangial cells expressed TNF-? mRNA and TNF-? protein at a high level. At the concentration of 1 ?g/L and 10?g/L, IL-13 markedly inhibited LPS-induced TNF-? mRNA and protein expression in mesangial cells. At the concentration of 100?g/L, IL-13 completely inhibited LPS-induced TNF-? mRNA and protein expression in mesangial cells. ICAM-1 constitutively expressed on the surface of mesangial cells at very low level. Induced by TNF-?(100?g/L), mesangial cells expressed ICAM-1 mRNA and ICAM-1 cell surface molecule at high level. Costimulated mesangial cells with IL-13(10?g/L) and TNF-?(100?g/L), IL-13 inhibited TNF-?-induced ICAM-1 mRNA and cell surface molecule expression at every time course. CONCLUSION: IL-13 not only inhibits LPS-induced TNF-? expression but also inhibits TNF-?-induced ICAM-1 expression in mesangial cells. These data suggest that IL-13 inhibits the inflammatory reaction in mesangial cells.
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AIM:To study the effect of human complement C 5b～9 complex on nitric oxide(NO) synthesis of glomerular mesangial cells (MC). METHODS: First, the human complement C 5b～9 complexes were isolated and glomerular MC of rats were cultured. Second, the MC were stimulated with C 5b～9 complex and changes of metabolism products of NO(NO 3 and NO 2) in MC culture supernatant at 6,24 and 48 hours after C 5b～9 stimulating were detected. Moreover, cGMP levels in cultured MC were also measured. RESULTS: NO 3/NO 2 contents from culture supernatant and cGMP levels in MC were increased parallelly after C 5b～9 complex stimulation. Further, NO synthesis was inhibited by L-NG-nitro-arginine-methylester(L-NAME). CONCLUSION: NO synthesis of rat glomerular MC was incerased by human complement C 5b～9 stimulation.