Role of serum neutrophil elastase determination in the diagnosis of acute exacerbation of asthma in preschool children / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the role of serum neutrophil elastase (NE) level in acute exacerbation of asthma in preschool children.</p><p><b>METHODS</b>A total of 85 preschool children who were diagnosed with asthma between January 2008 and January 2010 were classified into acute exacerbation group (n=44) and non-acute exacerbation group (n=41). Thirty-five children who received physical examination served as the control group. The enzyme-linked immunosorbent assay was used to determine the serum levels of NE and interleukin-8 (IL-8). The receiver operating characteristic (ROC) curve was used for NE evaluation.</p><p><b>RESULTS</b>Both the acute and non-acute exacerbation groups had higher serum levels of NE and IL-8 than the control group, and the acute exacerbation group had significantly higher serum levels of NE and IL-8 than the non-acute exacerbation group (P<0.05). The serum level of NE was positively correlated with that of IL-8 (r=0.48, P<0.05). With serum NE level >27.73 μg/L as the cut-off value for diagnosing acute exacerbation of asthma, the sensitivity was 65.9%, the specificity was 95.1%, and the area under the ROC curve was 0.87 (P<0.01).</p><p><b>CONCLUSIONS</b>The determination of serum NE level in preschool children with asthma helps to diagnose the acute exacerbation of asthma.</p>

Mechanisms of decorin inhibiting epithelial-to-mesenchymal transition induced by transforming growth factor beta1 in renal tubular epithelial cells / 中华儿科杂志

<p><b>OBJECTIVE</b>To investigate the mechanisms of decorin inhibiting epithelial-to-mesenchymal transition (EMT) induced by transforming growth factor beta1 (TGF-beta1) in renal tubular epithelial cells.</p><p><b>METHOD</b>HK-2 cells in vitro were divided into 4 groups: (1) negative control group; (2) decorin group, added with decorin 100 ng/ml ; (3) TGF-beta1 group, added with TGF-beta1 10 ng/ml; (4) decorin and TGF-beta1 group, added with decorin 100 ng/ml and TGF-beta1 10 ng/ml. The protein level of phosphor-ERK, phosphor-PI3K, phosphor-Smad(3) and beta-catenin was detected by Western blotting method. The snail mRNA level was tested by real time-PCR, while the lymphoid enhancer factor-1 (LEF-1) mRNA level was measured by RT-PCR.</p><p><b>RESULTS</b>The snail (2.59 +/- 0.70:1.02 +/- 0.13) and LEF-1 mRNA (1.85 +/- 0.08:0.30 +/- 0.11) were significantly up-regulated, meanwhile the protein level of phosphor-ERK (1.11 +/- 0.09:0.47 +/- 0.07), phosphor-PI3K (14.79 +/- 1.02:2.48 +/- 0.06), phosphor-Smad(3) (0.95 +/- 0.02:0.08 +/- 0.01) and beta-catenin (1.46 +/- 0.20:0.49 +/- 0.05) were significantly increased in TGF-beta1 group compared to control group, while there were no statistically significant difference in all figures between control group and decorin group. The phosphor-ERK protein level (0.58 +/- 0.08) and the snail mRNA level (1.24 +/- 0.03) were significantly down-regulated in TGF-beta1 and decorin group compared to TGF-beta1 group, however there were no statistically significant differences in the level of phosphor-PI3K (15.84 +/- 1.64), phosphor-Smad(3) (0.90 +/- 0.04) and beta-catenin (1.42 +/- 0.09) between these two groups.</p><p><b>CONCLUSION</b>Decorin inhibited EMT induced by TGF-beta1 which may be through blocking the ERK signal transduction pathway.</p>

Clinical and pathological features of Alport syndrome in children / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the clinical and pathological features of Alport syndrome in children.</p><p><b>METHODS</b>The clinical and histopathological data of 10 hospitalized children with Alport syndrome from February 2007 to February 2009 were retrospectively reviewed.</p><p><b>RESULTS</b>There were 7 males and 3 females, with the age ranging from 2 years to 6 years and 7 months (mean 3 years and 2 months). Five of 10 cases had positive family history. X-linked dominant inheritance Alport syndrome was diagnosed in 8 cases, and autosomal recessive inheritance Alport syndrome in 2 cases. Recurrent gross hematuria was found in 5 cases, hematuria and proteinuria in 3 cases, massive proteinuria in 1 case, and nephritic syndrome in 1 case. Under the light microscope, 8 cases presented with mesangial proliferation glomerulonephritis, and 2 cases with focal segmental glomerulosclerosis. Immunofluorescence assay showed that all cases had IgM deposition in glomerulus. Only 1 case showed typical glomerular basement membrane (GBM) pathological changes. All cases showed abnormal alpha-chain distribution in renal collagen IV.</p><p><b>CONCLUSIONS</b>The children with Alport syndrome have diverse clinical manifestations. Characteristic histopathological presentations could not be found under a light microscope, mesangial proliferation glomerulonephritis is the dominant pathological change, and IgM deposition in glomerulus is common. The GBM pathological change in children is not common. Immunofluorescence assay of alpha-chain in collagen IV is needed for the diagnosis of Alport syndrome.</p>

NADPH oxidase-derived reactive oxygen species involved in angiotensin II-induced monocyte chemoattractant protein-1 expression in mesangial cells / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the origin of oxidative stress induced by angiotensin II (AngII) in human mesangial cells and the role of reactive oxygen species (ROS) in AngII-induced monocyte chemoattractant protein-1 (MCP-1) expression.</p><p><b>METHODS</b>MCP-1 expression was determined by real time RT-PCR. ROS production was measured by DCFDA fluorescence. Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity was examined by lucigenin chemiluminescence. p47phox and p67phox translocation was assayed by Western blot. Twenty-four male mice were randomly divided into three groups: the control, the AngIIinfusion [AngII 400 ng/(kg.min)], and the apocynin treatment. AngII was infused by subcutaneously osmotic minipump for 14 days. Urinary albumin and 8-isoprostane excretion were measured by ELISA.</p><p><b>RESULTS</b>In cultured human mesangial cells, AngII induced the MCP-1 expression in a dose-dependent manner with 3.56 fold increase as compared with the control. AngII increased intracellular ROS production as early as 3 min with the peak at 60 min and was in a time and dose-dependent. Incubation with different dosages of AngII (1 nmol/L, 10 nmol/L, and 100 nmol/L AngII) for 60 min, ROS production increased at 1.82, 2.92, and 4.08 folds respectively. AngII-induced ROS generation was sensitive to diphenyleneiodonium sulfate (DPI, 10 micromol/L) and apocynin (500 micromol/L), two structurally distinct NADPH oxidase inhibitors. In contrast, inhibitors of other oxidant-producing enzymes, including the mitochondrial complex Iinhibitor rotenone, the xanthine oxidase inhibitor allopurinol, the cyclooxygenase inhibitor indomethacin, the lipoxygenase inhibitor nordihydroguiaretic acid, the cytochrome P450 oxygenase inhibitor ketoconazole and the nitric oxide synthase inhibitor G-nitro-L-arginine methyl ester were without an effect. AngII-induced ROS generation was inhibited by the AT1 antagonist losartan (10 micromol/L) but not the AT2 antagonist PD123319 (10 micromol/L). AngII treatment induced translocation of cytosolic of p47phox and p67phox to the membrane. The antioxidants almost abolished AngII-induced MCP-1 expression. AngII infusion increased urinary and p67 translocation by 2.69-, 2.97-, and 2.67-fold, respectively.</p><p><b>CONCLUSIONS</b>NADPH oxidase-derived ROS is involved in AngII-induced MCP-1 expression. Inhibition of NADPH oxidase alleviates AngII-induced renal injury.</p>

Effects of small interfering RNA (siRNA) against Par-4 gene on the apoptosis of human bone marrow mesenchymal stem cells / 中华儿科杂志

<p><b>OBJECTIVE</b>The prostate apoptosis response factor-4 (Par-4) gene was originally identified by differential screening for genes that are up-regulated when prostate cells are induced to undergo apoptosis. Par-4 was found to possess potent apoptotic activity in various cellular systems in response to numerous stimuli. The aim of this study was to explore the effects of small interfering RNA (siRNA) against Par-4 gene on the apoptosis of human bone marrow mesenchymal stem cells (hBMSCs) exposed to glutamate.</p><p><b>METHODS</b>Primary culture of hBMSCs was carried out and siRNAs targeted Par-4 gene (Par-4-SiRNA) were chemically synthesized. Eukaryocytic expression vector was built and were transfected into hBMSCs with liposome. After selecting with G418, the stable cell clones were treated with glutamate. The expression of Par-4 mRNA was determined by real-time PCR. The apoptosis of hBMSCs was quantified by flow cytometry. Western blotting was used to detect the protein levels of phosphorylated Akt1 (Thr308). Relative Caspase-3 activity was determined by colorimetric assay.</p><p><b>RESULTS</b>The Par-4-SiRNA-1 and Par-4-siRNA-2 could markedly down-regulate the mRNA levels of Par-4 gene in hBMSCs. With the transfections of Par-4-SiRNA-1 and Par-4-SiRNA-2, the levels of Par-4 mRNA were respectively decreased by 88% and 67%. Both Par-4-SiRNA-1 and Par-4-SiRNA-2 inhibited significantly the apoptosis of hBMSCs induced by glutamate, in which the percentages of apoptotic cells were respectively decreased to 38.80% +/- 3.97% (P < 0.01) and 45.49% +/- 4.32% (P < 0.01) from 60.30% +/- 6.82%. Western blot assays demonstrated that, glutamate down-regulated the expression of phosphorylated Akt1 proteins in hBMSCs (89.07 +/- 6.42 and 28.30 +/- 5.65, respectively, P < 0.01). However, Par-4-SiRNA-1 and Par-4-SiRNA-2 could markedly recover the down-regulation of Akt1 proteins induced by glutamate (63.56 +/- 6.75 and 45.59 +/- 4.88, respectively, P < 0.01). And the relative Caspase-3 activity which was enhanced by the treatment with glutamate (0.1428 +/- 0.0495 and 0.8616 +/- 0.1051, P < 0.01), was suppressed by Par-4-SiRNA-1 and Par-4-SiRNA-2 (0.8616 +/- 0.1051 and 0.6581 +/- 0.0555, respectively, P < 0.01).</p><p><b>CONCLUSION</b>SiRNA against Par-4 gene could inhibit the apoptosis of hBMSCs induced by glutamate, and its inhibitory effects may be mediated by the up-regulation of phosphorylated Akt1 and the suppression of the relative Caspase-3 activity.</p>

Comparison of urinary proteomics between steroid-sensitive and steroid-resistant minimal change nephrotic syndrome in children / 南方医科大学学报

<p><b>OBJECTIVE</b>To screen the differentially expressed proteins in the urine of children with steroid-sensitive and steroid-resistant minimal change nephrotic syndrome (SRINS and SSINS, respectively).</p><p><b>METHODS</b>Urine samples were collected from 10 children with SRINS and 70 with SSINS as well as 30 healthy volunteers (control). Isoelectric focusing and two-dimensional electrophoresis in combination with matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry was performed for analysis of the urine proteins.</p><p><b>RESULTS AND CONCLUSION</b>In the urine samples, 30 protein spots were identified to have differential expression between SRINS and SSINS. Further analysis of 14 protein spots identified 12 proteins expressing in SRINS, namely kinesin family member 27, PITPNB, bullous pemphigoid antigen, alpha-1 protease inhibitor, Zn-alpha-2GP, alpha-1B-glycoprotein, serum albumin precursor, haptoglobin precursor, kinesin like motor protein, IRAK4, cytoplasmic dynein and cytokeratin 9. Nine of these 12 proteins were up-regulated (U1-U3, U5, U7-U9, U11-U12) and 3 down-regulated (D4, D6, D10) in SRINS, suggesting that these proteins may serve as the potential therapeutic targets and as new diagnostic markers for steroid-resistant nephrotic syndrome.</p>

Association of Interleukin-1?-511C/T and Interleukin-1?+3953C/T Polymorphisms with Susceptibility of Pediatric Epilepsy / 实用儿科临床杂志

Objective To explore the relationship between IL-1?-511C/T and IL-1?+3953C/T site polymorphisms and the susceptibility of pediatric epilepsy.Methods Under the case-control study,IL-1?-511C/T and IL-1?+3953C/T site polymorphisms in 117 patients with pediatric epilepsy and 95 healthy individuals controls(healthy control group) were analyzed with polymerase chain reaction restriction and fragment length polymorphism(PCR-RFLP),the relationship between IL-1?-511C/T,IL-1?+3953 C/T site polymorphisms and the risk of pediatric epilepsy were analyzed.SAS 8.0 software was used to analyze the data.Results Multiple variate logistic regression analysis revealed that compared with healthy control group,there was no relationship between the IL-1?-511C/T site polymorphisms and the susceptibility of pediatric epilepsy individuals,carrying at least one +3953T variant allele(CT and TT genotypes) had a significantly increased risk for pediatric epilepsy(adjusted OR=2.46,95%CI 1.03-5.87),compared with the wild-type genotype(+3953CC).Furthermore,individuals with epilepsy or febrile seizures family history had a significantly higher risk(adjusted OR=4.12,95%CI 1.28-29.34),compared with those with both CC genotypes.Conclusions These findings support the hypothesis that IL-1?-511C/T site polymorphisms have no relationship with epilepsy,but the IL-1?+3953C/T polymorphism may contribute to the risk of developing pediatric epilepsy.

Changes of Aspartate and Glutamate in Hippocampus Formation and Cerebrospinal Fluid in Kainite Acid Induced Epilepsy Rat / 实用儿科临床杂志

Objective To explore the changes of contents of glutamate (Glu) and aspartate (Asp) in hippocampus formation and cerebrospinal fluid (CSF) in kainite acid(KA) induced epilepsy rats.Methods SD rats(n=40) were divided into 2 groups randomly:the KA group [intracerebroventricular injection(icv) of KA, 2 ?g/kg] and control group(icv of NS). KA group were divided into 4 groups at 6 h,1,3 and 7 d,each group 8 rats.High pressure liquid chromatgraphy(HPLC) was used to assess the concentrations of Glu and Asp in hippocampus formation and CSF.Results In the hippocampus, the contents of Glu and Asp increased continuously 1 d after seizure , but not different from those of control group.Three days later, only Glu became significantly different from control group. However, the contents of Glu and Asp in the CSF were significantly different from the control 6 h after seizure.Conclusion The contents of excite amino acid (especially Glu)in CSF increase immediately after KA injection, which are earlier than those in hippocampus formation.

Effect of Rosmarinic Acid on Antioxidative Systems in Mesangial Proliferative Glomerulonephritis in Rats / 实用儿科临床杂志

Objective To investigate the change of oxidation system and antioxidation system in mesangial proliferative glomerulonephritis(MsPGN) induced by anti-Thy1.1 antibody,and further to study the intervention of rosmarinic acid(RAD).Methods Anti-THy1.1 serum was produced,and then intravenously injected into rats for establishing an experimental model of MsPGN.The experiment was designed for control with or without RAD,glomerulonephritis with or without RAD,respectively.The activity of superoxide dismutase(SOD) and the content of malondialdehyde(MDA) in tissue homogenate were detected by spectrophotomerty.Results The activity of SOD significantly decreased,while the content of MDA increased in MsPGN.RAD could inhibit oxidation in the mesangial cells.Conclusion Lipid peroxidation participates in MsPGN and RAD can control the changes of the mesangial cells and show the activity of antioxidation.

The cloning of human Smac gene and its pro-apoptotic effect on Burkitt's lymphoma cells / 中华儿科杂志

<p><b>OBJECTIVE</b>Second mitochondria-derived activator of caspase (Smac) is a recently identified, novel pro-apoptotic molecule, which is released from mitochondria into the cytosol during apoptosis. Smac promotes activation of caspases by neutralizing members of the inhibitor of apoptosis proteins (IAPs) family, such as X-linked inhibitor of apoptosis protein (XIAP). The objective of the study was to examine the pro-apoptotic effect of human Smac gene on Burkitt's lymphoma Raji cells.</p><p><b>METHODS</b>The full length cDNA of human Smac gene was amplified by reverse transcription-PCR from total RNA of HEK-293 cells. The PCR product was ligated with linearized vector pGEM-T-easy supplied in the TA cloning kit and sequenced. The correct cDNA of full length Smac was subcloned into eukaryocytic expression vector pcDNA3.1/myc-his and transfected into human Burkitt's lymphoma cell line Raji by lipofectamine-mediated transfection. The expression of full length Smac was determined by Western blot. Morphological observation was done with the laser scanning confocal microscope by double staining the Raji cells with Hoechest 33,258 and propidium iodide. Flow cytometry was used to evaluate apoptosis. Relative caspase-3 activity was determined by colorimetric assay.</p><p><b>RESULTS</b>Recombinant eukaryocytic expression vector pcDNA3.1/Smac, which contained full length Smac, was successfully constructed. After pcDNA 3.1/Smac was transfected into human Burkitt's lymphoma Raji cell line for 24 hours, Raji cells showed apparent apoptosis with a percentage of (43.7 +/- 2.5)%, which was higher than that of non-transfected group and free vector-transfected group (P < 0.05). Compared with non-transfected group (0.136 +/- 0.036) and free vector-transfected group (0.138 +/- 0.026), the relative caspase-3 activity of Raji cells transfected by pcDNA3.1/Smac (0.936 +/- 0.041) was significantly enhanced (P < 0.05).</p><p><b>CONCLUSION</b>Transfection and expression of human Smac gene could significantly induce apoptosis of human Burkitt's lymphoma Raji cells. The mechanism is associated with the increase of caspase-3 activity.</p>

Suppressive effects of par-4 antisense oligodeoxynucleotide on up-regulation of intracellular calcium concentration in PC12 cell induced by glutamate and its anti-apoptosis effects / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the uppressive effects of par-4 antisense oligodeoxynucleotide on the up-regulation of intracellular calcium concentration in PC12 cell induced by glutamate and its anti-apoptosis effects.</p><p><b>METHODS</b>Cationic lipid-mediated par-4 antisense oligodeoxynucleotide (par-4-AS-ODN) was transfected into PC12 cells and followed by glutamate for treatment. Mismatch oligodeoxy-nucleotide (MS-ODN) was used as the control. Morphological assessment and evaluation of the anti-apoptosis effects of par-4-AS-ODN on PC12 cells were performed by laser scanning confocal microscopy by double staining of the cells with Hoechest 33258/propidium iodide (Hoe/PI) and flow cytometry respectively. The mRNA and protein levels of calpain 10 and par-4 were measured by RT-PCR and Western blot. Intracellular calcium concentration was determined by using laser scanning confocal microscope with Fura-2/AM as the fluorescent dye.</p><p><b>RESULTS</b>Par-4-AS-ODN repress the increase of par-4 protein in PC12 cell (52.3 +/- 5.0 vs 90.0 +/- 3.2, < 0.01). Par-4-AS-ODN significantly inhibited the apoptosis of PC12 cells induced by glutamate (53% vs 31%, < 0.01). Par-4-AS-ODN significantly suppress the up-regulation of intracellular calcium concentration in PC12 cells induced by glutamate (Rate of fluorescent density: 167.9 +/- 32.4 vs 228.8 +/- 36.8, < 0.01). Par-4-AS-ODN inhibited the increase of calpain 10 mRNA in PC12 cells induced by glutamate (46.3 +/- 3.7 vs 34.8 +/- 2.1, < 0.01).</p><p><b>CONCLUSIONS</b>par-4-AS-ODN enables to inhibit apoptosis of PC12 cells induced by glutamate. The mechanism of the inhibition may be closely related to suppression of the up-regulation of intracellular calcium concentration and calpain transcription expression.</p>

Transfection of Lipoxin A4 receptor-like protein gene enhanced the inhibitory effect of Lipoxin A4 on human lung fibroblasts proliferation induced by connective tissue growth factor / 中华儿科杂志

<p><b>OBJECTIVE</b>Lipoxin A(4) is formed by the metabolism of arachidonic acid. Anti-inflammatory and anti-proliferative effect of lipoxin A(4) has been shown in many human diseases. Recently, as a novel high affinity receptor for ligand lipoxin A(4), Lipoxin A(4) receptor-like protein (LRLP) has been identified. Currently close attention is paid to the important contribution of connective tissue growth factor (CTGF) in lung fibrosis. The purpose of the study was to transfect LRLP gene into human lung fibroblasts and investigate the mechanism of its enhancing antagonistic effect of Lipoxin A(4) on human lung fibroblasts proliferation induced by connective tissue growth factor.</p><p><b>METHODS</b>Eukaryocytic expression vector pEGFP/LRLP which contained LRLP and green fluorescence protein fusion gene (GFP) was constructed and transfected into human lung fibroblasts (HLF). After selecting with G418, HLF/LRLP cell clone which stably expressed LRLP/GFP fusion protein was isolated and characterized by the laser scanning confocal microscope. Cultured HLF and HLF/LRLP were stimulated for 24 h with CTGF (1 microg/ml) in the presence and absence of pretreatment of Lipoxin A(4) (10.0 nmol/L) for 30 min. Inhibition of cell proliferation was determined by MTT assay. Cell cycle analysis was performed by flow cytometry. Western blot was used to detect the expression of cyclin D(1) protein. Electrophoretic mobility shift assay (EMSA) was employed to detect the DNA binding activity of STAT(3).</p><p><b>RESULTS</b>(1) HLF/LRLP cell clone which stably expressed LRLP and GFP fusion protein was successfully obtained. (2) Proliferation of HLF and HLF/LRLP was induced by 1 microg/ml CTGF. Pretreatment with 10 nm Lipoxin A(4) inhibited the proliferation of HLF and HLF/LRLP. And the inhibitory rate of HLF/LRLP was significantly higher than that of HLF [(54.1 +/- 4.2)%, (21.2 +/- 3.7)%, P < 0.05]. (3) The flow cytometry analysis showed that compared with HLF, more HLF/LRLP were arrested at G(0)/G(1) phase in the presence of pretreatment of Lipoxin A(4). [(76.3 +/- 3.5)%, (60.8 +/- 2.0)%, P < 0.05]. (4) Ten nmol/L Lipoxin A(4) antagonized CTGF induced increase of cyclin D(1) protein expression in HLF and HLF/LRLP. And its antagonistic effect on HLR/LRLP was stronger than that on HLF (P < 0.05). (5) Ten nmol/L Lipoxin A(4) antagonized CTGF induced increase of STAT(3) DNA binding activity, and its antagonistic effect on HLF/LRLP was more powerful than that on HLF (P < 0.05).</p><p><b>CONCLUSIONS</b>Transfection of Lipoxin A(4) receptor-like protein gene enhanced the inhibitory effect of Lipoxin A(4) on human lung fibroblasts proliferation induced by CTGF. Its mechanism might be related to regulation of cyclin D(1) protein expression and STAT(3) DNA binding activity.</p>

Induction of monocyte chemoattractant protein-1 expression in human mesangial cells by angiotensin II: role of c-Jun N-terminal kinase-c-Jun/activator protein-1 signal pathway / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the role of c-Jun N-terminal kinase (JNK)-c-Jun/activator protein-1 (AP-1) signal pathway in expression of monocyte chemoattractant protein-1 (MCP-1) in experimental rat glomerulonephritis.</p><p><b>METHODS</b>Nephrotoxic sera nephritis (NTN) was induced by injection of anti-GBM antibody into the tail veins of rats. Electrophoretic mobility shift assay (EMSA) and non-radioactive kinase assay were used to detect the activity of AP-1 and JNK in kidneys and angiotensin II-stimulated human mesangial cells. Ribonuclear protection assay was used to detect MCP-1 expression in cultured human mesangial cells.</p><p><b>RESULTS</b>Significant up-regulation of JNK and AP-1 was observed in NTN rats (3.82 +/- 0.58) folds and (5.36 +/- 0.61) folds, as compared with the controls. Supershift assay demonstrated that c-Jun and c-Fos were the predominant subunits involved. Activation of JNK and AP-1 significantly correlated with MCP-1 expression in NTN rats. Angiotensin II enhanced the expression of MCP-1 and activation of JNK and AP-1 in cultured human mesangial cells in a dose-dependent manner, with maximal stimulation seen at 100 nmol/L (20.99 +/- 4.71) folds, (6.91 +/- 1.65) folds and (7.82 +/- 1.32) folds respectively. Significant down-regulation of AP-1 activation and MCP-1 expression were observed in angiotensin II-induced human mesangial cells pretreated with JNK specific inhibitor SP600125.</p><p><b>CONCLUSIONS</b>Angiotensin II and MCP-1 may play an important role in glomerulosclerosis via the JNK-c-Jun/AP-1 signal pathway.</p>

Expression of monocyte chemoattractant protein-1 in experimental rat glomerulonephritis is mediated by NF-kappaB/IkappaB signal pathway / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the role of NF-kappaB/IkappaB signal pathway in mediating the expression of monocyte chemoattractant protein-1 (MCP-1) in experimental rat glomerulonephritis.</p><p><b>METHODS</b>Nephrotoxic serum nephritis (NTN) was induced by injection of anti-GBM antibody into the tail veins of rats. Electrophoretic mobility shift assay (EMSA) and Western Blot were used to detect the activation of NF-kappaB, nuclear translocation of p65 subunit and degradation of IkappaBalpha and IkappaBbeta in rat renal tissue. MCP-1 expression in glomeruli and renal tubules was also assessed by immunohistochemistry and ribonuclease protection assay. This was further correlated with the activation of NF-kappaB.</p><p><b>RESULTS</b>There was an obvious expression of MCP-1 in glomeruli and renal tubules. Significant up-regulation of NF-kappaB activation, nuclear translocation of p65 subunit, and degradation of IkappaBalpha and IkappaBbeta were also observed in NTN rat renal tissue, as compared to the control group. A positive correlation was noted between NF-kappaB activation and MCP-1 expression.</p><p><b>CONCLUSIONS</b>NF-kappaB/IkappaB signal pathway may play an important pathogenetic role in glomerulonephritis, with mediating the expression of MCP-1.</p>

Curcumin inhibited the proliferation and extracellular matrix production of human mesangial cells / 中华儿科杂志

<p><b>OBJECTIVE</b>Glomerulosclerosis is characterized by extracellular matrix accumulative and is often associated with mesangial cell proliferation. Curcumin showed a protective effect on anti-glomerular basement membrane (anti-GBM) nephritis in vivo, although their cellular localization and mechanism of action is still unclear. In this study, a glomerular mesangial cell line derived from fetus was used to determine whether curcumin could inhibit the cell proliferation and alter the extracellular matrix turnover.</p><p><b>METHODS</b>The cell activity was determined with MTT method. Mesangial cells were cultured in vitro and incubated with 0, 3.125, 6.25, 12.5, 25, 50, 100 and 200 micromol/L curcumin. In addition,human mesangial cells were cultured with or without LPS (10 microg/ml) in presence or absence of various concentrations of curcumin (4, 16 and 200 micromol/L), respectively. The supernatant and cells were collected. Then, the levels of the collagen type IV and III protein in the supernatant were determined by using enzyme-linked immunosorbent assay and the IL-1 beta and MCP-1 mRNA in the cells was measured by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) after subconfluent quiescent mesangial cells were incubated with various concentrations of curcumin for 24 h in vitro.</p><p><b>RESULTS</b>Curcumin at the concentration equal to or over 6.25 micro mol/L was able to inhibit the proliferation of mesangial cells in a dose-dependent manner, the optical density according to the sequential concentrations of curcumin was 0.65 +/- 0.02, 0.62 +/- 0.04, 0.56 +/- 0.01, 0.53 +/- 0.02, 0.51 +/- 0.03, 0.44 +/- 0.05, 0.41 +/- 0.07 and 0.38 +/- 0.06. Without any stimulation, human mesangial cells secreted some collagen type IV and III (10 +/- 9.13 ng/ml and 29.5 +/- 0.58 ng/ml, respectively) and expressed some MCP-1 mRNA, but did not express IL-1 beta mRNA. LPS increased the expression of collagen type IV and III in the culture medium of mesangial cells in vitro [(138.75 +/- 23.23) ng/ml and (38.25 +/- 5.38) ng/ml] and up-regulated the IL-1 beta and MCP-1 mRNA expression [(16.91 +/- 1.68)% and (76.6 +/- 6.59)%]. Yet curcumin could significantly decrease collagen type IV and III in the supernatant of cultured mesangial cells induced by LPS (20.5 +/- 1.00, P < 0.05 and 20.5 +/- 4.12 ng/ml, P < 0.05) and down-regulated the mRNA expression of IL-1 beta and MCP-1 in mesangial cells induced by LPS (P < 0.01).</p><p><b>CONCLUSION</b>Curcumin could inhibit the human mesangial cell proliferation and alter the extracellular matrix turnover, meanwhile it could down-regulate the IL-1 beta and MCP-1 mRNA expression induced by LPS, which may be valuable in decreasing the progression of glomerulosclerosis.</p>