Biomedical subjects

Yao-Cheng Rui

Publications and source records attributed to Yao-Cheng Rui.

16 recordsLinked to original sources

PubMed2007

Proteomic analysis of foam cells.

Foam cells are characteristic pathological cells in the lesions of atherosclerosis. Previous works have established macrophage-derived foam cell model to study the central role of the foam cells, and analyzed the protein expression profiles in foam cells. The reported in vitro foam cell model was established by incubating the human U937 cells with oxidized low-density lipoprotein. The global changes in protein expressions between U937 foam cell and normal U937 cells were measured with two-dimensional gel electrophoresis, and some interested proteins were tryptic-digested and then identified via mass spectrometry after capillary liquid chromatography separation. Some of the identified proteins were validated via the Internet links to the U937 proteomic map provided from the Expasy Proteomics server (http://us.expasy.org). The experimental data can provide potential markers during the inflammatory reactions for atherosclerotic studies.

Chromatography, Liquid↗

PubMed2006

Different expressions of Nogo-B1 and Nogo-B2 in mouse heart microvascular endothelial cell dysfunction induced by lysophosphatidylcholine.

To investigate Nogo-B expression changes in mouse heart microvascular endothelial H5V cell line induced by lysophosphatidylcholine. Cells were incubated with different concentrations of lysoPC for the same incubation time and with 10 micromol/l lysoPC at different incubation times. Protein and mRNA expression levels of Nogo-B1 and Nogo-B2 were measured with Western blotting and semiquantitative RT-PCR, respectively. Nogo-B1 protein was detected in normal H5V cells by Western blotting. When H5V cells were incubated with lysoPC, Nogo-B1 protein level decreased, and the lowest point fell to 20% of the original level induced by 20 micromol/l lysoPC for 24 h. Incubation of H5V cells with lysoPC of different concentrations or at different time points caused little change in Nogo-B1 mRNA expression level, except for a 50% decrease in 20 micromol/l lysoPC at 24 h, while a transient change was observed in Nogo-B2 mRNA level. These results demonstrate that Nogo-B1 protein expression could be down-regulated with increasing concentrations of lysoPC and lapse of incubation time, though no mRNA transcription down-regulation occurred. However, mRNA expression level of Nogo-B2 showed a transient up-regulation induced by lysoPC. We conclude that the two subtypes of Nogo-B may play different roles in the endothelial cell injury process.

Animals↗

PubMed2006

C-reactive protein induced expression of adhesion molecules in cultured cerebral microvascular endothelial cells.

Ischemic stroke can trigger an acute phase response resulting in a rise of plasma concentration of C-reactive protein (CRP). Clinical data about the relationship between CRP and prognosis suggest that CRP might be involved in the pathogenesis of cerebral ischemia. In the present work, a significant increase of circulating level of CRP was observed in an vivo rat brain ischemia model of middle cerebral artery occlusion. To determine the possible effects of CRP on brain microvessel endothelium, we performed a dose-dependent experiment in mouse brain microvascular endothelial cells (bEnd.3 cells) with emphasis on its relation to cell adhesions molecules. Incubation with CRP (1-75 mg/L) for 24 h significantly increased Lactate dehydrogenase (LDH) leakage from bEnd.3 cells (P<0.01) in a dose-dependent manner, and induced significant up-regulations of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) expressions analyzed by Western blotting (P<0.01). In contrast to earlier report, CRP also induced significant increase in ICAM-1 expression in the absence of serum (P<0.01). In conclusion, the present results suggest that CRP may be involved directly in the development of inflammation in response to cerebral ischemia.

Animals↗

PubMed2005

Reduction of atherosclerosis in cholesterol-fed rabbits and decrease of expressions of intracellular adhesion molecule-1 and vascular endothelial growth factor in foam cells by a water-soluble fraction of Polygonum multiflorum.

Polygonum multiflorum stilbeneglycoside (PMS) is a water-soluble fraction of Polygonum multiflorum Thunb., one of the most famous tonic traditional Chinese medicines, that has protective effects on the cardiovascular system. The purpose of the present study is to elucidate the effects of PMS on macrophage-derived foam cell functions and the reduction of severity of atherosclerosis in hypercholesterolemic New Zealand White (NZW) rabbits. NZW rabbits were fed for 12 weeks with a normal diet, a high cholesterol diet, or a high cholesterol diet associated with irrigation with different doses of PMS (25, 50, or 100 mg/kg). Treatment of NZW rabbits fed with high cholesterol diet with 100 mg/kg PMS attenuated the increase in plasma cholesterol, low-density lipoprotein cholesterol, very low-density lipoprotein cholesterol, and plasma triglyceride. Treatment with 50 and 100 mg/kg PMS caused 43% and 60% decrease in atherosclerotic lesioned area ratio to total surface area, respectively. In U937 foam cells, PMS could decrease the high expression of intercellular adhesion molecule (ICAM)-1 protein and the vascular endothelial growth factor (VEGF) protein levels in the medium induced by oxidized lipoprotein when analyzed by flow cytometry. The results proved that PMS is a powerful agent against atherosclerosis and that PMS action could possibly be through the inhibition of the expression of ICAM-1 and VEGF in foam cells.

Animals↗

PubMed2005

Time courses of vascular endothelial growth factor and intercellular adhesion molecule-1 expressions in aortas of atherosclerotic rats.

An increasing number of studies have focused on the expressions of growth factors and adhesion molecules in atherosclerotic lesions, which are confirmed to play central roles in angiogenesis and endothelial dysfunction, including vascular endothelial growth factor (VEGF) and intercellular adhesion molecule-1 (ICAM-1). However, the difference of growth factor and adhesion molecule expression time courses has not been determined in vivo. This study aimed to determine the expression patterns and expression curves of ICAM-1 and VEGF in atherosclerotic rats during the time course. An experiment atherosclerotic model in rats was established by combining the high fat/cholesterol diets with injection of vitamin D3. In situ hybridization was used to determine the expression patterns of VEGF and ICAM-1 in aortas of normal or atherosclerotic rats in 8 weeks. There was a massive increase in reactivity for both ICAM-1 and VEGF in atherosclerotic plaques. Northern blot, Western blot and ELISA analysis were used to quantify VEGF and ICAM-1 expressions in time course. In rat aorta, the expression curves in time course showed that ICAM-1, not VEGF, was up-regulated in mRNA levels significantly in 2 weeks; while VEGF expression was hysteresis than ICAM-1, which showed maximum expression level in 8 weeks. Our results provide the evidence of VEGF and ICAM-1 expression curves in time course in atherosclerotic rats, which indicated different regulatory mechanisms of VEGF and ICAM-1 expression in atherogenesis.

Animals↗

PubMed2005

Expression of pro-inflammatory and anti-inflammatory cytokines in brain of atherosclerotic rats and effects of Ginkgo biloba extract.

AIM: To study the protein and mRNA expressions of pro-inflammatory and anti-inflammatory cytokines in the brain of rats with atherosclerosis (AS) and the effects of Ginkgo biloba extract (GbE) on expressions of cytokines. METHODS: The experimental model of AS in rats were established by intraperitioneal injection of vitamin D3 with high fat/cholesterol diet. GbE 100 mg/kg was administered to rats by ig. After 8 weeks, the expressions of IL-1beta, TNF-alpha, IL-10, and IL-10R in the brain tissues of AS rats were detected by enzyme-linked immunosorbant assay, immunohistochemistry, Western blotting, and reverse transcriptase polymerase chain reaction. RESULTS: The protein and mRNA expressions of IL-1beta, TNF-alpha, and IL-10 in the brains were markedly higher in AS groups than that in control groups (6.11+/-0.15, 1.55+/-0.14, 0.54+/-0.04 ng/g wet weight vs 0.80+/-0.14, 0.33+/-0.09, and 0.33+/-0.02 ng/g wet weight, respectively). The protein and mRNA expressions of IL-1beta and TNF-alpha in the brains were markedly lower in GbE groups (3.82+/-0.54, 0.95+/-0.08 ng/g wet weight) than that in AS groups, the protein and mRNA expressions of IL-10 and IL-10R in the brains were markedly higher in GbE groups (0.85+/-0.06 ng/g wet weight) than that in AS groups. CONCLUSION: GbE inhibited production of pro-inflammatory cytokines IL-1beta and TNF-alpha, but up-regulated the production of anti-inflammatory cytokines, IL-10 and IL-10R in brain, which might be related with its anti-AS actions.

Animals↗

PubMed2004

Comparative analysis of the proteome of left ventricular heart of arteriosclerosis in rat.

Despite the worldwide occurrence of coronary atherosclerotic heart disease (CAHD), the pathogenic mechanisms underlying this disease remain largely unknown. In this study, the experimental model of atherosclerosis in rat (CAHD rat) was established by the injection of vitamin D3 associated with high fat diet for 6 weeks. By using the proteomic approach, we comparatively analyzed the proteome of the control and CAHD rat left ventricular myocardial tissues. We reproducibly separated over 2500 polypeptides by using two-dimensional electrophoresis (2-DE) at pH range of 3-11. Among these proteins, 26 proteins with large amount were identified using micro high performance liquid chromatography mass spectrometer/mass spectrometer (micro-HPLC-MS/MS). Using PDQUEST software to process the 2-DE gel images, 38 protein spots that significantly altered in CAHD were detected. Of these, 12 proteins were identified with high confidence by using 2-DE and matrix-associated laser desorption ionization time-of-flight mass spectrometer (MALDI-TOF-MS). The identification of protein alterations specify to CAHD would clarify the pathogenetic mechanisms involved in the disease and might be of prognostic and therapeutic benefit.

Animals↗

PubMed2004

Analysis of gene expression profiles in human HL-60 cell exposed to cantharidin using cDNA microarray.

Cantharidin is a natural toxin that has antitumor properties and causes leukocytosis as well as increasing sensitivity of tumor cells resistant to other chemotherapeutic agents. There is limited information, however, on the molecular pharmacological mechanisms of cantharidin on human cancer cells. We have used cDNA microarrays to identify gene expression changes in HL-60 promyeloid leukemia cells exposed to cantharidin. Cantharidin-treated cells not only decreased expression of genes coding for proteins involved in DNA replication (e.g., DNA polymerase delta), DNA repair (e.g., FANCG, ERCC), energy metabolism (e.g., isocitrate dehydrogenase alpha, ADP/ATP translocase), but also decreased expression of genes coding for proteins that have oncogenic activity (e.g., c-myc, GTPase) or show tumor-specific expression (e.g., phosphatidylinositol 3-kinase). In contrast, these treated cells overexpressed several genes that encode intracellular and secreted growth-inhibitory proteins (e.g., BTG2, MCP-3) as well as proapoptotic genes (e.g., ATL-derived PMA-responsive peptide). Our findings suggest that alterations in specific genes functionally related to cell proliferation or apoptosis may be responsible for cantharidin-mediated cytotoxicity. We also found that exposure of HL-60 cells to cantharidin resulted in the decreased expression of multidrug resistance-associated protein genes (e.g., ABCA3, MOAT-B), suggesting that cantharidin may be used as an oncotherapy sensitizer, and the increased expression of genes in modulating cytokine production and inflammatory response (e.g., NFIL-3, N-formylpeptide receptor), which may partly explain the stimulating effects on leukocytosis. Our data provide new insight into the molecular mechanisms of cantharidin.

Cantharidin↗

PubMed2004

[Effect of madecassoside on depression behavior of mice and activities of MAO in different brain regions of rats].

OBJECTIVE: To evaluate the effects of madecassoside (MC) on the depression behavior of mice and the activities of monoamine oxidase (MAO) in different rat brain regions. METHODS: Imipramine as the positive contrast medicine, effects of MC on the depression behavior of mice were observed by forced swimming test and reserpine antagonist test. Moclobemide and pargyline as the positive controlled medicines, the activities of monoamine oxidase-A (MAO-A) and monoamine oxidase-B (MAO-B) in different rat brain regions were determined after intragastric administration of MC in 3 different dosages for 3 days or 21 days. RESULTS: (1) The low, middle and high dosages of MC (i.g.) significantly reduced the immobility time of mice in forced swimming test (P<0.05). (2) MC in dosages of 10 mg/kg and 20 mg/kg prevented the lowering of temperature induced by reserpine (P<0.05), while 40 mg/kg had no significant effects on it (P>0.05). (3) With acute administration (3 days), the low, middle and high dosagey of MC (i.g.) significantly inhibited the activity of MAO-A in hippocampus (P<0.01), and the high dosage significantly inhibited the activity of MAO-A in hypothalamus (P<0.01), while the 3 dosages had no significant effects on the activity of MAO-A in cortex (P>0.05). With chronic administration (21 days), MC in 3 dosages had no significant effects on the activities of MAO-A in cortex and hypothalamus (P>0.05), and the high dosage (40 mg/kg) significantly enhanced the activity of MAO-A in hippocampus (P<0.01). (4) With acute administration, MC in dosages of 10 mg/kg and 20 mg/kg significantly inhibited the activity of MAO-B in cortex (P>0.05), and MC in dosage of 10 mg/kg significantly inhibited the activity of MAO-B in hypothalamus (P<0.05), and MC in dosage of 20 mg/kg significantly enhanced the activity of MAO-B in hippocampus (P<0.01). With chronic administration, MC of 3 dosages produced no significant effects on the activities of MAO-B in 3 different rat brain regions (P>0.05). CONCLUSION: These results support the idea that MC produces antidepressant effects through MAO inhibition in rat brain, which seems stronger with acute administration than chronic administration, while its mechanism remains to be further studied.

Animals↗

PubMed2004

Inhibitory effect of extracts of Ginkgo biloba leaves on VEGF-induced hyperpermeability of bovine coronary endothelial cells in vitro.

AIM: To study whether extract of Ginkgo biloba (EGb) can protect against atherosclerosis. METHODS: Confluent monolayers of bovine coronary endothelial cells (BCECs), bovine coronary smooth muscle cells (BCSMCs), and cocultures of the two were incubated with medium containing VEGF and/or EGb, and flux of 125I-labeled oxidized low density lipoprotein (ox-LDL) across the monolayers was measured. RESULTS: Incubation with VEGF significantly increased the permeability of BCEC monolayers to 125I-ox-LDL in a time- and concentration-dependent manner, but had no effect on permeability of BCSMCs or endothelial cells-smooth muscle cells cocultures. EGb significantly inhibited the VEGF-induced hyperpermeability of BCECs. CONCLUSION: VEGF was important in the formation and development of atherosclerosis. The inhibition of VEGF-induced permeability by EGb suggests that extracts of Ginkgo biloba leaves may have important clinical applications in the treatment of cardiovascular diseases.

Animals↗

PubMed2003

Intercellular adhesion molecule-1 and vascular endothelial growth factor expression kinetics in macrophage-derived foam cells.

Macrophage-derived foam cells seem to play an important role during inflammatory response of atherosclerosis, in which the overexpression of intercellular adhesion molecule-1 (ICAM-1) and vascular endothelial growth factor (VEGF) are associated with the early and later pathological changes in foam cell formation. In this study, we investigated the expression kinetics of ICAM-1 and VEGF in macrophage-derived foam cells. The foam cell model was established through incubating the human monocyte line (U937 cells) with oxidized-low density lipoprotein (ox-LDL). Up-regulated expressions of ICAM-1 and VEGF were analyzed in protein and mRNA levels in U937 foam cells by flow cytometry, ELISA, and Northern blot. Kinetic studies showed the deferent kinds of expression curves in dose response and time course. The expression dose-kinetics demonstrated that the ICAM-1 showed the peak expression induced by ox-LDL 50 mg/L, while VEGF levels increased in a dose-dependent manner with the maximum level induced by ox-LDL 200 mg/L. Time-kinetic studies revealed that the ICAM-1 levels showed the peak expression in 12 h while VEGF expression increased in a time-dependent manner with the maximum level in 48 h. These results proved that both ICAM-1 and VEGF expressions were enhanced in the macrophage-derived foam cells, but ICAM-1 expression increased earlier than the up-regulation of VEGF; low dose of ox-LDL mainly up regulated ICAM-1 expression, while high dose mainly increased the VEGF expression.

Blotting, Northern↗

PubMed2003

Proteomic studies of macrophage-derived foam cell from human U937 cell line using two-dimensional gel electrophoresis and tandem mass spectrometry.

SUMMARY: The authors have identified 37 proteins in the whole-cell extracts from human monoblastic leukemia (U937) cell and macrophage-derived foam cell. The in vitro foam cell model was established by incubating the human U937 cells with oxidized low-density lipoprotein. The global changes in protein expressions between U937 foam cell and normal U937 cells were measured with two-dimensional polyacrylamide gel electrophoresis, and some proteins were trypsin-digested and then identified through tandem mass spectrometry after capillary liquid chromatography separation. Some of the identified proteins were validated via Internet links to the U937 proteomic map provided on the Expasy proteomics server. The experimental data can provide potential markers for atherosclerotic studies.

Electrophoresis, Gel, Two-Dimensional↗

PubMed2003

Antisense oligodeoxynucleotide inhibits vascular endothelial growth factor expression in U937 foam cells.

AIM: To study the expression of vascular endothelial growth factor (VEGF) induced by oxidized low density liporotein (ox-LDL) and the inhibitory effects of antisense oligodeoxynucleotide (asODN) on the levels of VEGF protein and mRNA in the U937 foam cells. METHODS: U937 cells were incubated with ox-LDL 80 mg/L for 48 h, then, the foam cells were treated with asODN (0, 5, 10, and 20 micromol/L). The VEGF concentration in the media was determined by ELISA. The VEGF protein expression level in cells was measured by immuohistochemistry; the positive ratio detected by a morphometrical analysis system was used as the amount of the VEGF expression level. The VEGF mRNA level was examined by Northern blotting. RESULTS: After U937 cells were incubated with ox-LDL, VEGF expression level increased greatly both in the cells and in the media. asODN markedly inhibited the increase of VEGF. After treatment with asODN 20 micromol/L, the VEGF protein concentration in the media decreased by 45.0%, the VEGF positive ratio detected by immuohistochemistry in cells decreased by 64.9%, and the VEGF mRNA level decreased by 47.1%. CONCLUSION: The expression of VEGF in U937 foam cells was strong. asODN inhibited VEGF expression significantly in U937 foam cells in vitro.

Foam Cells↗

PubMed2003

Protein expressions in macrophage-derived foam cells: comparative analysis by two-dimensional gel electrophoresis.

AIM: To study the overall protein expression changes induced by oxidized low-density lipoprotein (ox-LDL) in the U937 foam cells. METHODS: Foam cell model was established by incubating the human monoblastic leukemia (U937) cells with ox-LDL. Each protein samples in U937 control cells and U937 foam cells were separated by two-dimensional gel electrophoresis (2-DE). After the gels were stained by silver staining method, the images were analyzed by PDquest 2D-image-analysis software (Bio-Rad). Some of the spots were available via the Internet with links to the U937 proteomic map provided from the ExPASy Proteomics server. RESULTS: Using 2-DE, the overall protein map was obtained, in which 150 spots were matched with the control gel (match ratio: 75 %). Compared with U937 cells, 37 spots significantly changed in the foam cells (P<0.05), among which the expression levels in 28 spots increased and those in 9 spots decreased. Especially, 8 spots in U937 cells were absent in the foam cells, while 11 spots in the foam cells were absent in the control cells. CONCLUSION: The changed protein profiles induced by ox-LDL in U937 foam cells were established to support the functional studies on the macrophage-derived foam cells in atherosclerotic pathological states.

Cells, Cultured↗

PubMed2002

Expression of intercellular adhesion molecule-1 in U937 foam cells and inhibitory effect of imperatorin.

AIM: To investigate the expression level of intercellular adhesion molecule-1 (ICAM-1) in a macrophage-derived foam cell model from human U937 cell line and the inhibitory effect of imperatorin (IMP) on the ICAM-1. METHODS: U937 cells were incubated with oxidized low density lipoprotein (ox-LDL) 80 mg/L for 48 h and a macrophage-derived foam cell model was established. The medium was pretreated with different concentrations of IMP (0, 25, 50, 100 micromol/L). ICAM-1 protein expression in cells was measured with Western blotting; ICAM-1 mRNA level in cells was measured by Northern blotting. RESULTS: After incubated with ox-LDL, ICAM-1 expression level increased greatly. The increase in ICAM-1 protein level and mRNA level was estimated to be about 15-fold and 10-fold. When the cells were pretreated with imperatorin (50, 100 micromol/L), the increase of ICAM-1 in foam cells were remarkably inhibited. Especially when pretreated with IMP 100 micromol/L, the ICAM-1 protein level decreased by 79 % and the mRNA level decreased by 74 % each compared to the level of foam cells. CONCLUSION: After incubated with ox-LDL in vitro, the U937 foam cells showed an enhanced ICAM-1 expression compared with normal U937 cells. IMP could inhibit the expression of ICAM-1 in U937 foam cells.

Angelica↗

PubMed2002

Inhibitory effects of Ginkgo biloba extract on vascular endothelial growth factor in rat aortic endothelial cells.

AIM: To study the protective effects of Ginkgo biloba extract (GbE) against rat aortic endothelial cells (RAEC) damage induced by lysophosphatidylcholine (LPC). METHODS: Cell injury were determined by MTT assay and LDH release. Vascular endothelial growth factor (VEGF) protein production from RAEC was determined by enzyme-linked immunosorbent assay (ELISA). VEGF mRNA expression was examined by in situ hybridization and dot blot. RESULTS: GbE 0.01-1 microg/L prevented LPC-induced injury in cultured RAEC in a concentration-dependent manner. Cultured RAEC could express VEGF protein and VEGF mRNA was induced by LPC 5 mg/L. GbE could inhibit the expression of VEGF protein and VEGF mRNA in co-cultured RAEC with LPC. CONCLUSION: LPC could induce a strong expression of VEGF in RAEC. GbE could protect RAEC against the LPC-induced damage and downregulate VEGF protein and VEGF mRNA expression in cultured RAEC.

Animals↗