2.Rosiglitazone Inhibited Aldosterone-Induced Mesangial Cell Proliferation via Blocking Phosphatidyl Inositol 3-Kinase Activation
kang-kang, XU ; ai-hua, ZHANG ; gui-xia, DING
Journal of Applied Clinical Pediatrics 2006;0(17):-
Objective To explore the inhibitory effect of rosiglitazone of peroxisome proliferator-activated receptor-?(PPAR?) agonist on aldosterone-induced mesangial cell(MC) proliferation.Methods Mouse primary MC were cultured and treated with aldosterone(100 nmol/L) in the presence or absence of rosiglitazone(1.0,2.5,5.0,10.0 ?mol/L).The incorporation of 3H-thymidine(3H-TdR) and cell count were used as the measure of MC proliferation.Cyclin D1 and cyclin A expression,PI3K and Akt phosphorylation were determined by Western blot analysis.Results 1.Aldosterone induced MC proliferation,as assessed by 3H-TdR incorporation and cell number,which were increased by 2.46-and 2.14-fold,respectively,in aldosterone-treated cells.Aldosterone-induced MC proliferation was inhibited by PPAR? agonist rosiglitazone in dose-dependent manner in mouse MC.2.Aldosterone induced cyclin D1 and cyclin A expression.Rosiglitazone reduced aldosterone-induced cyclin D1 and cyclin A expression in dose-dependent manner.3.Aldosterone induced PI3K/Akt activation in dose-dependent manner,incubation with 100 nmol/L aldosterone for 60 min,phosphorylation PI3K and Akt expression increased by above 3.0-fold.4.PI3K inhibitor LY294002 and Akt inhibitor significantly inhibited aldosterone-induced cyclin D1 and cyclin A expression.5.Rosiglitazone significantly inhibited aldosterone-induced PI3K/Akt activation,10 ?mol/L rosiglitazone almost completely blocked aldosterone-induced PI3K/Akt activation.Conclusion Rosiglitazone can block aldosterone-induced MC proliferation via inhibition of PI3K/Akt activation.
3.Effect of SP600125 on AngⅡ-Induced Transforming Growth Factor-1 and Fibronectin Expression in Human Mesangial Cells
kang-kang, XU ; ai-hua, ZHANG ; gui-xia, DING
Journal of Applied Clinical Pediatrics 2006;0(23):-
Objective To investigate the effect of c-Jun N-terminal kinase(JNK) specific inhibitor SP600125 on Angiotensin Ⅱ(AngⅡ)-induced transforming growth factor-?1(TGF-?1) and fibronectin (FN) expression in human mesangial cells (MC).Methods Human MC were isolated and cultured in vitro and were treated with AngⅡ in the presence or absence of JNK specific inhibitor SP600125.The protein was isolated or the supernate of medium was collected at the end of experiment.JNK,extracellular signal-regulated kinase(ERK1/2),and p38 mitogen-activated protein kinase(MAPK) activity were determined by Western blot method.TGF-?1 and FN were determined by enzyme linked immunosorbent assay(ELISA).Results SP600125 inhibited AngⅡ-induced Ser63 phosphorylation of c-Jun in a concentration-dependent manner,and JNK activity was reduced by 75% at 10 ?mol/L and by 90% at 20 ?mol/L.SP600125 had no effect on AngⅡ-induced ERK1/2 and p38 activity.TGF-?1 and FN protein were constitutively produced in MC,and production was significantly stimulated for 8 to 48 h after addition of AngⅡ.Preincubation of cells with SP600125(20 ?mol/L) significantly inhibited AngⅡ-induced TGF-?1 and FN production during this time period.SP600125 inhibited AngⅡ-induced production of TGF-?1 and FN in a concentration-dependent manner.Conclusion SP600125 inhibited AngⅡ-induced JNK activation and TGF-?1 and FN expression in human MC and may serve as the novel approach for the treatment of patients with chronic kidney disease.
4.Effect of Rosmarinic Acid on Antioxidative Systems in Mesangial Proliferative Glomerulonephritis in Rats
lin, LI ; song-ming, HUANG ; san-long, ZHAO ; quan-cheng, FENG ; gui-xia, DING
Journal of Applied Clinical Pediatrics 2006;0(15):-
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.
5.Progress in study of selective ERβ ligands.
Jin-ya CAI ; Jun-hao LI ; Shi-hui DING ; Juan ZHANG ; Gui-xia LIU ; Wei-hua LI ; Yun TANG
Acta Pharmaceutica Sinica 2015;50(6):658-667
Estrogen receptors (ERs) are members of nuclear receptors and related to several diseases such as cancer, inflammation and osteoporosis. ERs have two forms, ERα and ERβ, which have different functions and organism distributions. Compounds selectively targeting ERβ can regulate important physiological functions and avoid the side effects caused by targeting ERα. Therefore, selective ERβ ligands have received considerable research interest in recent years. In this article, different kinds of selective ERβ ligands were summarized and their structure-activity relationships were also analyzed.
Estrogen Receptor beta
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chemistry
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Humans
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Ligands
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Structure-Activity Relationship
6.Effects of exogenous connective tissue growth factor on collagen III synthesis of human renal tubular epithelial cells.
Ai-Qing ZHANG ; Wei-Hua GAN ; Gui-Xia DING ; Jing GONG
Chinese Journal of Contemporary Pediatrics 2008;10(2):188-190
OBJECTIVETo explore the role of exogenous connective tissue growth factor (CTGF) in the collagen III synthesis of human renal tubular epithelial cell line HK2 in vitro.
METHODSCultured HK2 cells were randomly assigned to three groups: placebo-control, low-dose CTGF-treated (2.5 ng/mL) and high-dose CTGF-treated groups (20 ng/mL). Cell morphological changes were observed under an inverted microscope. Collagen III alpha mRNA expression was detected using RT-PCR. Immunohistochemistry staining was used to assess the levels of intracellular collagen III alpha protein.
RESULTSAfter 48 hrs of low- or high- dose CTGF treatment, the appearances of HK2 cells were changed from oval to fusiform. High-dose CTGF treatment increased collagen III alpha mRNA expression (0.4461+/-0.0274 vs 0.2999+/-0.0115; P<0.05) as well as the protein expression of collagen III alpha (0.4075+/-0.0071 vs 0.3503+/-0.0136; P<0.05) compared with the placebo-control group.
CONCLUSIONSCTGF can induce morphological changes of human renal tubular epithelial cells in vitro. High concentration of CTGF may increase the synthesis of collagen III alpha.
Cells, Cultured ; Collagen Type III ; biosynthesis ; genetics ; Connective Tissue Growth Factor ; Epithelial Cells ; drug effects ; metabolism ; Humans ; Immediate-Early Proteins ; pharmacology ; Immunohistochemistry ; Intercellular Signaling Peptides and Proteins ; pharmacology ; Kidney Tubules ; drug effects ; metabolism ; RNA, Messenger ; analysis
7.Clinical evidence of therapy of IgA nephropathy.
Wei-Hua GAN ; Ai-Qing ZHANG ; Gui-Xia DING ; Jing GONG
Chinese Journal of Contemporary Pediatrics 2007;9(2):101-103
Angiotensin-Converting Enzyme Inhibitors
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therapeutic use
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Anticoagulants
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therapeutic use
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Cyclophosphamide
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therapeutic use
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Cyclosporine
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therapeutic use
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Fish Oils
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therapeutic use
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Glomerulonephritis, IGA
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therapy
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Glucocorticoids
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therapeutic use
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Humans
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Mycophenolic Acid
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analogs & derivatives
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therapeutic use
8.Effects of nanosized titanium dioxide on the reproductive system of male mice.
Li-li GUO ; Xiao-hui LIU ; Ding-xia QIN ; Li GAO ; Hong-mei ZHANG ; Jia-yin LIU ; Yu-gui CUI
National Journal of Andrology 2009;15(6):517-522
OBJECTIVETo investigate the in vivo effects of nanosized titanium dioxide (TiO2) on the main organs, particularly the reproductive system, of male mice.
METHODSForty-five male ICR mice aged 6 weeks were equally and randomly divided into 2 experimental groups and a control group, intraperitoneally injected with 200 and 500 mg/kg nanosized TiO2 and equal volume of saline, respectively, every other day for 5 times. One week after drug cessation, we obtained the coefficients of the main organs, serum biochemical parameters and the levels of gonadal hormones (T and E2), analyzed the pathological changes of the main organs by HE staining, observed sperm count, motility and abnormality under the microscope, and detected germ cell apoptosis by TUNEL.
RESULTSCompared with the control, the low-dose (200 mg/kg) group showed no significant changes in the above parameters, while the high-dose (500 mg/kg) group exhibited a decrease in the coefficients of the liver, heart and kidneys, and a significant increase in such serum biochemical parameters as ALT, ALT/AST and BUN (P < 0.05). The high-dose group also showed significantly reduced sperm density and motility, increased sperm abnormality and germ cell apoptosis (P < 0.05), but no obvious pathological changes in the liver, kidney spleen, testis and epididymis.
CONCLUSIONLow-dose nanosized TiO2 has no obvious effect on male mice, but high-dose may significantly reduce their sperm count and function and induce germ cell apoptosis, although its damage on the liver and kidney function is slight.
Animals ; Apoptosis ; Male ; Mice ; Mice, Inbred ICR ; Nanoparticles ; Sperm Count ; Sperm Motility ; drug effects ; Spermatozoa ; drug effects ; Titanium ; pharmacology
9.NADPH oxidase-derived reactive oxygen species involved in angiotensin II-induced monocyte chemoattractant protein-1 expression in mesangial cells.
Ying CHEN ; Ai-hua ZHANG ; Song-ming HUANG ; Gui-xia DING ; Wei-zhen ZHANG ; Hua-ying BAO ; Hong-mei WU ; Rong-hua CHEN
Chinese Journal of Pathology 2009;38(7):456-461
OBJECTIVETo 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.
METHODSMCP-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.
RESULTSIn 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.
CONCLUSIONSNADPH oxidase-derived ROS is involved in AngII-induced MCP-1 expression. Inhibition of NADPH oxidase alleviates AngII-induced renal injury.
Acetophenones ; pharmacology ; Angiotensin II ; administration & dosage ; pharmacology ; Angiotensin II Type 1 Receptor Blockers ; pharmacology ; Animals ; Cells, Cultured ; Chemokine CCL2 ; metabolism ; Dose-Response Relationship, Drug ; Humans ; Losartan ; pharmacology ; Male ; Mesangial Cells ; metabolism ; Mice ; Mice, Inbred C57BL ; NADPH Oxidases ; antagonists & inhibitors ; metabolism ; Onium Compounds ; pharmacology ; Oxidative Stress ; Phosphoproteins ; metabolism ; Protein Transport ; Random Allocation ; Reactive Oxygen Species ; metabolism
10.Clinical and immunological features of lupus nephritis in children: retrospective analysis of 40 cases.
Xue-Qin CHENG ; Hua-Ying BAO ; Ai-Hua ZHANG ; Wei-Zhen ZHANG ; Gui-Xia DING ; Fei ZHAO ; Ying CHEN
Chinese Journal of Contemporary Pediatrics 2014;16(1):53-57
OBJECTIVETo analyze the clinical and immunological features of children with lupus nephritis (LN).
METHODSChart records of 40 (4 male and 36 female) LN children who were admitted consecutively between January, 2005 and December, 2010 were reviewed. The baseline demographic, pathological and immunological data were analyzed.
RESULTSIn the 40 LN patients analyzed, the mean age of the disease onset was 10.6 ± 2.6 (range from 2.6 to 14.3) years, and 35 cases (88%) were school-age children. Proteinuria was detected in all 40 cases, including nephrotic-range proteinuria in 12 (30%) cases, and isolated proteinuria in 9 (22%) cases. Twenty-six (65%) patients had varying degrees of hematuria. Acute nephritis was the most common sub-type, accounting for 47% of the total cases. Among the 39 cases undergoing renal biopsy, 3 were unclassified and the remaining 36 were classified, respectively, as type IV LN (50%, 18 cases), type II LN (22%, 8 cases). In the histopathologcally classified case, 100% were antinuclear antibody-positive, 61% were anti-dsDNA-positive, and 89% showed varying degrees of decrease in serum C3 and C4 concentrations. Following treatment for 6 months, a high LN remission rate (95%) was achieved; the acute renal activity index remained higher in IV, V+III and V+IV subtypes than in other subtypes, while the chronic index and the degree of tubulointerstitial damage were not different between histopathological subtypes.
CONCLUSIONSThe clinical manifestations of LN children are diverse. Clinically, acute nephritis is the most common form of LN in children. Histopathologically, type IV is the most frequent subtype of LN. Early treatment may result in significant disease remission.
Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Lupus Nephritis ; drug therapy ; immunology ; pathology ; Male ; Retrospective Studies