Angiotensin II ; pharmacology ; Blotting, Western ; Cells, Cultured ; Glomerular Mesangium ; cytology ; drug effects ; metabolism ; Humans ; I-kappa B Kinase ; NF-kappa B ; analysis ; Peptide Fragments ; pharmacology ; Protein-Serine-Threonine Kinases ; analysis

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.

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.

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.

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 ; chemistry ; Humans ; Ligands ; Structure-Activity Relationship

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

Angiotensin-Converting Enzyme Inhibitors ; therapeutic use ; Anticoagulants ; therapeutic use ; Cyclophosphamide ; therapeutic use ; Cyclosporine ; therapeutic use ; Fish Oils ; therapeutic use ; Glomerulonephritis, IGA ; therapy ; Glucocorticoids ; therapeutic use ; Humans ; Mycophenolic Acid ; analogs & derivatives ; therapeutic use

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

OBJECTIVETo observe the influence of therapy with Chinese medicine Lirukang Granule (, LRKG) combined with psychological intervention on anxiety states and sex hormones in patients with cyclomastopathy and menoxenia.

METHODSA total of 470 subjects were randomly assigned to three groups by the net-central randomization system, the treatment group (161 patients, treated with LRKG and psychological intervention), the Chinese medicine group (157 patients, treated with LRKG), and the psychological intervention group (152 patients, treated with psychological intervention). The dose of LRKG was 12 g three times per day; psychological intervention included establishing relations, cognitive intervention and psychological persuasion, 30-40 min per session, once a week. The therapy duration for all groups was three months. The efficacy was compared and anxiety state/State-Trait Anxiety Invertory (STAI) scoring was measured before and after treatment. The serum estradiol (E2), progesterone (P), prolactin (PRL) and follicle stimulating hormone (FSH) levels of 60 patients selected randomly from each group during the luteal phase were measured before and after treatment, and a group of 20 healthy women were evaluated for comparison. A follow-up was arranged for one year after treatment.

RESULTSThirty subjects were lost to follow-up. (1) Comparison of efficacy: the markedly effective rate and the total effective rate of the treatment group were 86.67% (131/150) and 98.00% (147/150), respectively; of the Chinese medicine group, 64.58% (93/144) and 90.27% (130/144), respectively; and of the psychological intervention group, 0% (0/146) and 3.42% (5/146), respectively. The markedly effective rate and the total effective rate in the treatment group were significantly higher than those in the Chinese medicine and psychological intervention groups (P < 0.05). (2) Comparison of STAI scoring: STAI scoring was decreased dramatically in the treatment group after treatment compared with that of the Chinese medicine group (P < 0.01), but there was no significant difference compared with the psychological intervention group. (3) Comparison of levels of sex hormones: E2, P, PRL and FSH of the three patient groups were disordered before treatment, and significantly different from healthy women (P < 0.01). After treatment, the levels of P and FSH of the treatment group were significantly increased (P < 0.01), E2 and PRL were significantly reduced, which were also significantly decreased compared with the psychological intervention groups (P < 0.01). (4) FOLLOW-UP: the markedly effective rate and the total effective rate of the treatment group remained higher than those of the other two groups after one year of treatment (P < 0.05). (5) Adverse reactions: no obvious adverse reactions were found among the three groups.

CONCLUSIONSTherapy with Chinese medicine combined with psychological intervention was effective for short-term and long-term treatment of cyclomastopathy and menoxenia. The mechanism might be related to the regulation of sex hormones.

Adult ; Behavior Therapy ; methods ; Breast Diseases ; physiopathology ; therapy ; Combined Modality Therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Menstrual Cycle ; Menstruation Disturbances ; physiopathology ; therapy ; Middle Aged ; Prospective Studies ; Psychotherapy ; methods ; Risk Assessment ; Treatment Outcome ; Young Adult

OBJECTIVETo 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.

METHODSNephrotoxic 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.

RESULTSSignificant 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.

CONCLUSIONSAngiotensin II and MCP-1 may play an important role in glomerulosclerosis via the JNK-c-Jun/AP-1 signal pathway.

Angiotensin II ; pharmacology ; Animals ; Cells, Cultured ; Chemokine CCL2 ; metabolism ; Glomerular Mesangium ; cytology ; metabolism ; Glomerulonephritis ; metabolism ; Humans ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Male ; Proto-Oncogene Proteins c-fos ; metabolism ; Proto-Oncogene Proteins c-jun ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Transcription Factor AP-1 ; metabolism