共查询到20条相似文献,搜索用时 15 毫秒
1.
AIM: To study and compare the pathophysiological effects of serum IgA1 from both the patients with IgA nephropathy (IgAN) and healthy controls on human mesangial cells (HMC). METHODS: Serum IgA1 was isolated with Jacalin affinity chromatography and heated to form aggregates (aIgA1). Primary HMC were cultured and passage 3 and passage 4 of the cells were used. Intracellular calcium release was assayed with confocal analysis. Expression of TGF-β mRNA and the content of supernatant fibronectin were tested by RT-PCR and indirect competitive ELISA, respectively. RESULTS: aIgA1 from patients with IgAN was shown to induce release of intracellular calcium, up-regulation of expression of TGF-β mRNA and secretion of fibronectin in HMC in a similar time-dependent manner as aIgA1 from healthy controls, but the effects of the former were much stronger and the durations was much longer (P<0.05, respectively). CONCLUSION: IgA1 from patients with IgAN was shown to be able to stimulate HMC with a stronger biological effects than that from healthy controls. It is suggested that direct interaction between IgA1 from patients with IgAN and HMC may be another mechanism in the pathogenesis of IgAN. 相似文献
2.
3.
AIM: To investigate whether transferrin receptor (TfR) and Fc α/μ R are the major IgA1 receptor on human mesangial cells (HMC). METHODS: Serum IgA1 was isolated by jacalin affinity chromatography and heated to aggregated form (aIgA1). RT-PCR was performed to investigate the expression of TfR mRNA and Fcα/μR mRNA. Binding capacity of aIgA1 to human mesangial cell line (HMCL) was evaluated by radio-ligand binding assay. Binding specificity was determined by competitive inhibition assay and phosphorylation of extracellular signal-regulated kinase (ERK) was determined by Western blot. RESULTS: TfR cDNA and Fcα/μR cDNA products were amplified from HMC but not from HMCL. aIgA1 was found to bind to HMCL in a dose-dependent, saturable manner and the binding was inhibited by BSA. Scatchard analysis revealed a Kd of (6.4±1.7)×10-7 mol/L and the binding sites were (3.0±1.2)×106/cells. Both aIgA1 from patients with IgAN and healthy controls were able to induce the phosphorylation of ERK in a similar time-dependent manner, but the effect of aIgA1 from patients with IgAN was much stronger (P<0.01) and the duration was much longer (P<0.05) than those from healthy controls. CONCLUSION: There might be a novel IgA1 receptor on HMCL, but TfR and Fc α/μ R are not the major candidates. 相似文献
4.
AIM: To investigate the role of heat-shock protein 70(HSP70) in the protection of myocardial cells against ischemic injury.METHODS: Myocardial cells were cultured in vitro. HSP70 was induced by hyperthermia. H2O2-injured myocardial cells were divided into different groups. Flow cytometry, DNA Ladder and biochemistry methods were employed to observe the myocardial cells of different groups.RESULTS: Immunohistochemistry showed hyperthermia induced the up-regulation of HSP70 in myocardial cells. Apoptotic rate, activity analysis of cytochrome C and succinic dehydrogenase in H2O2-injuried and HSP70-protected groups were obviously different. Electron micrograph shomed hyperthermia alliviated myocardial cell injury induced by H2O2. CONCLUSION: HSP70 delays apoptosis and protects against H2O2-induced myocardial cell injury. 相似文献
5.
AIM: To investigate the effect of IL-10 on IL-1β-induced prostaglandin E2(PGE2) release and cyclooxygenase-2(COX-2) expression in human mesangial cells and to examine whether IL-10 has effect on the biological function of IL-1β.METHODS: The PGE2 concentration in supernatants of HMC was measured by radioimmunoassay. The COX-2 mRNA and protein were measured by RT-PCR and Western blot, respectively. RESULTS: PGE2 and COX-2 were significantly increased after treatment with IL-1β(P<0.01 for both) in cultured human mesangial cells. IL-10 had no effects on basical production of COX-2 and PGE2(P>0.05, respectively), while it inhibited IL-1β-elicited PGE2 production, as well as COX-2 mRNA and protein expression in a concentration-dependent fashion. CONCLUSIONS: These results indicated that IL-10 depressed the IL-1β-induced release of PGE2 and expression of COX-2. These data suggested that IL-10 could exert anti-inflammatory actions at several levels, not only by inhibiting the production of pro-inflammatory cytokines but also by suppressing their biological function. 相似文献
6.
ZHANG Yi-min YE Ren-gao LI You-ji LI Xiao-yan WANG Chang-yun YU Xue-qing DONG Xiu-qing 《园艺学报》2007,23(9):1796-1800
AIM: To study the effects and mechanism of recombinant human defensin α1 on cell proliferation in cultured rat glomerular mesangial cells.METHODS: The influences of defensin α1 at various concentrations on rat 1097 mesangial cell line cultured in vitro were evaluated with MTT assay.The different concentrations of U0126,signal-regulated protein kinase (MEK) inhibitor,were added into the culture mediums of mesangial cells to do blocking test.Incubated with a final concentration of 3 mg/L defensin α1,the phosphorylation of extracellular signal regulated kinase (ERK)1/2 and type IV collagen of mesangial cells in different times were evaluated by Western blotting.RESULTS: Defensin α1 at 3-20 mg/L enhanced proliferation of rat glomerular mesangial cells.The incubation times for the maximum effect on proliferation was 12 h (P<0.01),whereas defensin α1 concentration >20 mg/L decreased cell proliferation.The cell proliferation induced by defensin α1 was inhibited by U0126.Stimulation of the cells with defensin α1 at concentration of 3 mg/L for 5 minutes induced a maximum effect on a ratio of phosphorylation of ERK1/2 to total ERK.After 12 h incubation with defensin α1,an increase in type IV collagen was observed by Western blotting and continued to increase at 24 h and 48 h (P<0.01).CONCLUSION: Defensin α1 enhances rat glomerular mesangial cell proliferation and induces type IV collagen production by MAPK signaling pathway. 相似文献
7.
WANG Ying-wei HE Qiu-zao QIN Hui-lian TANG Ren-xian GAO Feng-guang ZHANG Guang-yi 《园艺学报》2000,16(3):262-265
AIM:To study the effect of human complement C5b~9 complex on nitric oxide(NO) synthesis of glomerular mesangial cells (MC). METHODS: First, the human complement C5b~9 complexes were isolated and glomerular MC of rats were cultured. Second, the MC were stimulated with C5b~9 complex and changes of metabolism products of NO(NO3- and NO2) 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: NO3-/NO2- contents from culture supernatant and cGMP levels in MC were increased parallelly after C5b~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 C5b~9 stimulation. 相似文献
8.
AIM: To investigate the method of inducing G1 phase synchronization in human endometrial cancer JEC Cells by lovastatin and the cell cycle progress of JEC cells after desynchronization. METHODS: The doubling time of JEC cells was detected by Cell Counting Kit-8 (CCK-8) assay. To determine the best lovastatin concentrations for G1 synchronization, JEC cells were treated with lovastatin at concentrations of 10, 20, 30 and 40 μmol/L for 1× doubling time, and the cell cycle was detected using flow cytometry (FCM). To determine the best period of lovastatin treatment to achieve G1 synchronization, JEC cells were treated with lovastatin at the best concentration for 0.5× to 2× doubling time, and the cell cycle was detected every 4 h using FCM. Furthermore, the cell cycle progress of JEC cells after desynchronization was also observed. RESULTS: The doubling time of JEC cells was almost 24 h. Treatment with lovastatin at the concentration of 20 μmol/L for 24 h achieved maximum G1 arrest in JEC cells. Minimum G1 phase and maximum S phase were observed after desynchronization for 16 h. CONCLUSION: Maximum G1 synchronization of JEC cells is induced by lovastatin at the concentration of 20 μmol/L for 24 h. The JEC cells show minimum G1 phase and maximum S phase after desynchronization for 16 h. 相似文献
9.
AIM: To investigate the effects of the sera from the rats after limb ischemic preconditioning (LIPC) on human umbilical vein endothelial cells (HUVECs) injured by hydrogen peroxide (H2O2). METHODS: The HUVECs were divided into 5 groups: the cells in control group were cultured without any intervention; the cells in model group (M) were damaged by 1 mmol/L H2O2 for 2 h; the cells in early preconditioning serum (EPS) group, delayed preconditioning serum (DPS) group or sham limb ischemic preconditioning serum (SPS) group were treated with the corresponding serum at 5% for 12 h, respectively, and then treaed with H2O2 for 2 h. The viability of the HUVECs was analyzed by flow cytometry. The lactate dehydrogenase (LDH) in the culture media was detected. The cell adhesion molecules in the HUVECs were detected by real-time PCR. The mRNA and protein expression of heme oxygenase-1 (HO-1) was also determined. RESULTS: The viability of HUVECs incubated with 1 mmol/L H2O2 for 2 h significantly decreased compared with the control cells, which was accompanied with the augmentations of LDH in the medium and the cell adhesion molecules in cells, such as vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1). Preincubation with EPS and DPS derived from the rats subjected LIPC attenuated these injuries. Furthermore, pretreatment with EPS and DPS increased the expression of HO-1 at mRNA and protein levels. CONCLUSION: LIPC protects the HUVECs from H2O2-induced injury. 相似文献
10.
AIM: To explore the effects of lipoxin A4 on the expression of cyclooxygenase 2 (COX-2) in human bronchial epithelial cells (HBECs). METHODS: HBECs were incubated with various concentrations (0.1, 1 and 10 mg/L) of lipopolysaccharide(LPS) for 9 h, or 1 mg/L LPS for different time (3 h, 6 h and 9 h). The levels of COX-2 mRNA in HBECs and prostaglandin E2 (PGE2) in the culture supernatant were measured. In addition, the HBECs were exposed to lipoxin A4 at concentration of 0, 100 and 400 μmol/L after stimulated with LPS at concentration of 1 mg/L for 9 h, and the supernatant of the culture cells was collected for determining the content of PGE2 by ELISA. The cells were also harvested, and the mRNA and protein levels of COX-2 were analyzed by RT-PCR and Western blotting, respectively. RESULTS: LPS increased the mRNA expression of COX-2 and production of PGE2 in a dose and time dependent manners in HBECs. Induction of COX-2 mRNA and protein by LPS were inhibited by lipoxin A4 in a dose-dependent manner. Lipoxin A4 also significantly decreased LPS-induced production of PGE2. CONCLUSION: Lipoxin A4 down-regulates LPS-induced expression of COX-2 and consequently inhibits the production of PGE2 in HBECs. 相似文献
11.
AIM: To observe the expression of transforming growth factor β1 (TGF-β1), MAPK1/3 and fibronectin (FN) in the development of renal tubulointerstitial disease. METHODS: Wistar male rats were randomly divided into normal control group, diabetic group of 1week, 2 weeks, 4 weeks and 8 weeks. Diabetic model was induced by peritoneal injection of streptozotocin. Immunohistochemistry was employed to detect the expression of TGF-β1, MAPK1/3 and FN in the kidney. TGF-β1 protein in the renal cortex was checked by Western blot. BG, Scr and UP were analysed by biochemical methods, and the morphological changes in renal tubulointerstitium were also examined under microscopy on sections stained with HE and PAS. RESULTS: The expression of MAPK1/3 and FN was observed, but not the expression of TGF-β1 in normal renal tissue. Positive staining of TGF-β1 was observed in the renal tubulo-interstitium in 1-week diabetic group and thereafter it increased in the course of diabetes. A continuous increase in the expression of MAPK1/3 and FN was also observed in two - week diabetic rats. Chronologically the expression of TGF-β1,MAPK1/3 and FN and the ratio of KW/BW were positively correlative with each other in diabetic animals except one -week diabetic rats. There was also a positive correlation between MAPK1/3 and FN in l -week diabetic rats. CONCLUSION: Our data suggest that TGF-β1 appears in the renal tubulointerstitium in early period of diabetes and then its signal is mediated by MAPK1/3 cascades to accelerate production of FN ,and in turn leads to renal hypertrophy and tubulointerstitial fibrosis. 相似文献
12.
AIM: To investigate the effects of 1,25-dihydroxyvitamin D3 on the proliferation of passively-sensitized human airway smooth muscle cells (HASMCs), and to explore its potential role in asthmatic airway remodeling.METHODS: HASMCs were passively sensitized with 10% serum from asthmatic patients.1,25-(OH)2D3 was used as the interventor.The effect of 1,25-(OH)2D3 on the cell proliferation and its optimal concentration were determined by MTT colorimetric assay.The cell cycle analysis was performed by flow cytometry.The expression of proliferating cell nuclear antigen (PCNA) was measured by the method of immunocytochemical staining.RESULTS: 1,25-(OH)2D3 at the concentrations of 10-9-10-7 mol/L markedly inhibited the cell proliferation and the maximum effect was observed at the concentration of 10-7 mol/L.This concentration of 1,25-(OH)2D3 markedly suppressed the PCNA-positive rate and hampered the G1/S transition in HASMCs passively-sensitized by asthmatic serum.CONCLUSION: 1,25-(OH)2D3 has direct inhibitory effects on the proliferation of passively-sensitized HASMCs in vitro, which may be concerned with the beneficial role of 1,25-(OH)2D3 on the prevention and therapy of asthmatic airway remodeling. 相似文献
13.
WANG Hui-yan ZHANG Xiang-hong YAN Xia WANG Jun-ling SUN Xu-ming YANG Yong-bin WANG Feng-rong 《园艺学报》2001,17(2):131-133
AIM: To explore the effects of aflatoxin G1(AFG1 )on proliferation and TNF-α secretion of human peripheral blood mononuclear cells(HPBM) in vitro. METHODS: The effects of AFG1 on proliferation of HPBM were analysed with flow cytometric (FCM) DNA analysis and MTT bioassay, while that on TNF-α secretion was detected with ELISA.RESULTS: FCM analysis revealed that 6 h after treatment, proliferation index(PI) of 1000 μg/L AFG1 treated HPBM was significantly higher than that of control. 24 h after AFG1 treatment, stimulating effects on proliferation was found in HPBM treated with AFG1 at 200 μg/L and 1 000μg/L.Regression analysis showed that PI was postively correlated with the concentrations of AFG1 in the concentration range from 0 to 1 000μg/L( r=0. 5122 and 0.5119 respectively,P<0.05).MTT bioassay showed that the A value of the cells treated with AFG1 at 2 000 μg/L was higher than that of the control. Double antibody sandwich enzyme linked immunosorbent assay (ELISA) results showed that AFG1 at a dose of 100 μg/L could significantly inhibit lipopolysaccharide-induced TNF-α secretion.CONCLUSION: AFG1 could stimulate the proliferation of HPBM and could decrease TNF-α secretion at certain concentration. 相似文献
14.
AIM: To investigate the effects of Schisandrin B (Sch B) on apoptosis of lens epithelial cells (LEC) treated with H2O2. METHODS: Eyes in SDrats were excised and lenses were separated under operating microscope in sterilized condition. Lenses were divided randomly into four groups with different treatment: control group, hydrogen peroxide group (H2O2), pirenoxine sodium group (PS) and schisandrin Bgroup (Sch B). Lenses were incubated in CO2 incubator for 24 h with 300 μmol·L-1 H2O2 and with or without 0.5 mmol·L-1 Sch B. LECaoptosis and apoptosis rate were measured by TUNELmethod. Ultrastructure changes and apoptosis bodies of LECwere observed via transmitted electron microscope. RESULTS: (1) Apoptosis rate in H2O2 group (92.0±2.6) was significantly higher than that in control group (3.5±1.8). Apoptosis rate in Sch Bgroup (13.8±3.27) was remarkably lower than that in H2O2 group and PSgroup. (2) Ultrastructure observation indicated that apoptosis cells occurred in most LEC in H2O2 group and the changes were severe presenting different stages. While a few apoptosis cells were observed in Sch Bgroup, the changes were slight and most of them were in early and middle stages. CONCLUSION: These data indicated that Sch Bsignificantly inhibited apoptosis of LECduring experimental oxidative injury, the effects were stronger than PS. 相似文献
15.
16.
AIM: To study the changes of serum autoantibodies against β1-adrenergic and M2-muscarinic receptors in obstructive sleep apnea syndrome (OSAS) patients with chronic obstructive pulmonary disease (COPD), that is, overlap syndrome (OS). METHOD: Serum autoantibodies against β1 and M2 receptors in 26 cases with OS, 32 with OSAS, 30 with COPD and 28 normal subjects were determined by using enzyme-linked immunosorbent assay(ELISA). RESULTS: The positive rates and titer of β1 and M2 receptor autoantibodies are significantly increased in OS group (92.2%,57.7% and 1:98, 1:67), compared to OSAS (71.9%, 40.6% and 1:83, 1:30) or COPD group (70.0%, 36.7% and 1:79, 1:28) (P<0.05), and they are higher in these groups than in the control (25.0%, 14.3% and 1:20, 1:20) (P<0.01). CONCLUSION: Serum β1-and M2 receptor autoantibodies are significantly increased in the patients with COPD, OSAS or OS, compared to the control, and the highest is in OS. 相似文献
17.
AIM: To explore the mechanism of ET-1, NO and PGI2 release from coronary artery endothelial cells(CAEC) induced by acute hypoxia. METHODS: Bovine coronary artery endothelial cells were cultured and [45 Ca2+] was used to investigate the difference of calcium uptake between normoxia group and hypoxia group(3% O2). The contents of ET-1, NO and PGI2 in media of normoxia group, hypoxia group and hypoxia + verapamil group were measured 24 h after hypoxia. RESULTS: [ 45 Ca2+] uptake by CAEC in hypoxia group was 1.9 times more than normoxia group(P< 0.01). Hypoxia + verapamil group released more PGI2, ET-1 and less NO than hypoxia group(P< 0.05). CONCLUSION: Changes of ET-1, NO and PGI2 releases during hypoxia may be caused by the inflow of Ca2+ into coronary artery endothelial cells. 相似文献
18.
19.
AIM:To prepare m1AChR-G11 and m4AChR-G16 fusion protein in Baculovirus-Sf9 cell system and detect the effects of various muscarinic ligands on the interaction between m1AChR and G11 and m4AChR and G16, and screen different kinds of ligands specific for m1 and m4. METHODS:To prepare fused DNA of m1AChR-G11and m4AChR-G16 in two PCR, then expressed in Sf9 cells and detect the pharmacological function of m1AChR-G11 fusion protein and m4AChR-G16 fusion protein by QNB and GTPγS binding experiments; To expore the way of the activation of m1AChR-G11 and m4AChR-G16 fusion protein by various ligands includingcetylcholine (ACh), Pilocarpine (Pilo), 4-hydroxy-2-butynyl-1-trimethylammonium-m-chloro-carbanilatechloride (McN-A-343), tetrandrine, pirenzepine (PZ), alcuronium, atropine, R-(+)-hyoscyamine and gallamine by displacement by GDP on GTPγS binding experiments. RESULTS:The expression levels of m1AChR-G11 and m4AChR-G16 fusion protein were (45.39±2.62) nmol·g-1 protein, (47.04±1.58) nmol·g-1 protein. The affinity of GDP to G11 and G16 partner changed in the presence of different muscarinic ligands. CONCLUSION: The m1AChR-G11 and m4AChR-G16 showed the pharmacological specificity to m1 and m4 receptor and the efficient signaling of the two partners. Ligands of m1AChR and m4AchR mediated different signal transduction by changing the affinity of G11/G16 and GDP. So m1AChR-G11 fusion protein and m4AChR-G16 fusion protein can be taken as a tool to screen ligands specific for m1AChR and m4AChR. 相似文献
20.
AIM: To investigate the synergistic effect of decitabine (DCA) and valproic acid (VPA) on apoptosis and cell cycle arrest at G0/G1 phase in gastric cancer MGC-803 cells. METHODS: Gastric cancer MGC-803 cells were used in the study and divided into the following groups according to the treatment with different drugs for 72 h: DCA 1.5 μmol/L,DCA 3.0 μmol/L, VPA 1.5 mmol/L, DCA 1.5 μmol/L+VPA 1.5 mmol/L and DCA 3.0 μmol/L+VPA 1.5 mmol/L. The early and late apoptotic rates were detected by annexin V and PI staining. The cell cycle was also determined by flow cytometry. The relative nm23-H1 mRNA expression level was measured by real-time quantitative PCR. RESULTS: The apoptotic rates in VPA 1.5 mmol/L+DCA 1.5 μmol/L group (early: 33.58%±3.88%; late: 31.52%±4.20%) and VPA 1.5 mmol/L+DCA 3.0 μmol/L group (early: 42.61%±4.23%; late: 38.01%±3.86%), the percentages of the cells in G0/G1 phase in VPA 1.5 mmol/L+DCA 1.5 μmol/L group (61.55%±2.38%) and VPA 1.5 mmol/L+DCA 3.0 μmol/L group (66.75%±2.48%), and the relative nm23-H1 mRNA expression levels in VPA 1.5 mmol/L +DCA 1.5 μmol/L group (1.84±0.46) and VPA 1.5 mmol/L+DCA 3.0 μmol/L group (3.02±0.36) were all significantly higher than those in the corresponding concentrations of single drug treatment groups (P<0.01). CONCLUSION: Synergistic effect of VPA and DCA on apoptosis and cell cycle arrest in gastric cancer MGC-803 cells is possibly via inactivation of nm23-H1 gene expression. 相似文献