The aim of the study was to investigate the effect of black soybean (BS) on the susceptibility of low-density lipoprotein (LDL) in hypercholesterolemic New Zealand white rabbits. Effects of the BS extract (BSE) and it...The aim of the study was to investigate the effect of black soybean (BS) on the susceptibility of low-density lipoprotein (LDL) in hypercholesterolemic New Zealand white rabbits. Effects of the BS extract (BSE) and its components on monocyte adhesion of human aortic endothelial cells (HAECs), and adhesion molecule were investigated. Rabbits were divided into four groups, including control, 0.5% cholesterol with 20% casein (either with or without 0.5% vitamin E), and BS groups, all fed for 8 weeks. LDL was treated with 10 μM Cu2+ in vitro to determine the LDL lag time, and the vitamin E content of LDL was determined. The thickness of the tunica intima was measured on paraffin sections of thoracic aortas and aortic arches stained with Movat’s pentachrome. HAECs were pretreated with 100 μg/ml of BSE, and 10 μM of genistein, daidzein, cyanidin, and aspirin for 18 h, followed by tumor necrosis factor (TNF)-α (2 ng/ml) for 6 h, after which U937 cell adhesion was determined. Adhesion molecule expression was examined using ELISAs. The LDL lag time in the BS group was similar to that in the vitamin E group, while its lag time was significantly longer than those in the control and casein groups. The ratio of the intimal area/medial area of the aortic arch of the casein group was significantly higher than those in the control, BS, and vitamin E groups. The vitamin E group had the lowest value, and was closest to the control group. The BS group exhibited a significantly decreased atheroma region in the aortic arch compared to the casein group. Pre-incubation with BSE, genistein, daidzein, cyanidin, and aspirin significantly decreased adhesion by U937 monocytic cells to TNF-α stimulated HAECs. Genistein, daidzein, cyanidin, and aspirin significantly suppressed the expression of vascular cell adhesion molecule (VCAM)-1. Only genistein and aspirin significantly decreased intracellular adhesion molecule (ICAM)-1 expression compared to TNF-α treatment, while no treatments had any effect on E-selectin expression. BS significantly prolonged the LDL lag time and decreased the atheroma region of the aortic arch in hypercholesterolemic rabbits, thereby exerting an antiatherosclerotic effect. Presumably, the BSE downregulate intracellular redox-dependent signaling pathways in HAECs upon TNF-α stimulation through regulating NF-κB, thereby attenuating the inflammatory response in atherosclerosis. The antiatherogenic and anti-inflammatory effects of BS can be used as a nutraceutical for atherogenesis prevention.展开更多
[Objectives] To study the anti-inflammatory activity and mechanism of salvianolic acid A(Sal A), salvianolic acid C(Sal C), and rosmarinic acid(RA) on HAECs induced by TNF-α. [Methods] VE was used as a positive contr...[Objectives] To study the anti-inflammatory activity and mechanism of salvianolic acid A(Sal A), salvianolic acid C(Sal C), and rosmarinic acid(RA) on HAECs induced by TNF-α. [Methods] VE was used as a positive control and TNF-α induced human aortic endothelial cells(HAECs) were selected as the inflammation model cells. The levels of IL-6, MCP-1 were determined using ELISA. The mRNA levels of IL-6, MCP-1, ICAM-1, P-selectin and NF-κB were analyzed by quantitative RT-PCR. The protein expression of NF-κB was determined by western blot. The adhesion of U937 to HAECs was assessed by BCECF/AM labeling assay. Immunohistochemistry detection was used for ICAM-1 and P-selectin expression levels in TNF-induced HAECs, and DHE detection for the ROS production in HAECs induced by TNF-α. [Results] TNF-α-induced over-expressions of IL-6, MCP-1, NF-κB, ICAM-1 and P-selectin in HAECs were down-regulated by Sal A, Sal C and RA both in mRNA or protein levels, respectively. Meanwhile Sal A, Sal C and RA could significantly inhibit the production of ROS and U937 cells adhesion to HAECs. [Conclusions] Phenolic acids could inhibit TNF-α-induced inflammatory responses on HAECs and the mechanism might be related to inhibition of the production of ROS and blocking NF-κB signaling pathway.展开更多
Objective To investigate the expression of endoplasmic reticulum stress(ESR)marker C/EBP homologous protein-10(CHOP-10)in the human aortic endothelial cells(HAEC)under the ischemia and hypoxia stress and to study the ...Objective To investigate the expression of endoplasmic reticulum stress(ESR)marker C/EBP homologous protein-10(CHOP-10)in the human aortic endothelial cells(HAEC)under the ischemia and hypoxia stress and to study the effects of atorvastatin on the process.Methods The cultured HAEC were divided into normal control group,ischemia/hypoxia model group。展开更多
Cellular immune response is a major barrier to xenotransplantation, and cell adhesion is the first step in intercellular recognition. Flow-cytometric adhesion assay has been used to investigate the differential adhesi...Cellular immune response is a major barrier to xenotransplantation, and cell adhesion is the first step in intercellular recognition. Flow-cytometric adhesion assay has been used to investigate the differential adhesions of monocyte (Mo), natural killer celi (NK) and T lymphocyte (T) present within human peripheral blood mononuclear cells (PBMC) to porcine aortic endothelial cells (PAEC), and to demonstrate the effect of human interferon-γ (hIFN-γ) or/and tumor necrosis factor-α (hTNF-α) pretreatment of PAEC on their adhesiveness f or different PBMC subsets. The preferential sequence for PBMC subset binding to resting PAEC is Mo, NK and T cells, among which T cells show the slightest adherence; hTNF- a can act across the species, and augment Mo, NK and T cell adhesion ratios by 40%, 110% and 3 times, respectively. These results confirm at the cell level that host Mo and NK cells are major participants in the cellular xenograft rejection, thereby, providing a prerequisite for further studying the展开更多
Using human umbilical vein endothelial cells (HUVEC) and porcine aortic endothelial cells (PAEC) as target cells, human peripheral blood NK cells (PBNK) and NK92 cells as effector cells, the differential cytotoxicitie...Using human umbilical vein endothelial cells (HUVEC) and porcine aortic endothelial cells (PAEC) as target cells, human peripheral blood NK cells (PBNK) and NK92 cells as effector cells, the differential cytotoxicities of NK cells to allo- and xeno-endothelial cells were studied. The influence of MHC class I molecules on the cytotoxicity of human NK cells was assayed using acid treatment, and blockades of MHC class I antigens, CD94 and KIR (NKB1). The results indicated that the killing of PAEC by the two kinds of NK cells is higher than that of HUVEC. After acid-treatment, the cytotoxicity of the two kinds of NK cells to PAEC and HUVEC is significantly enhanced, but the magnitude of the enhancement is different. The enhancement of NK killing to acid treated HUVEC is much greater than that to PAEC. Blockade of CD94 mAb did not alter the NK cytotoxicity, while blockade of NKB1 mAb enhanced the cytotoxicity of PBNK to HUVEC and PAEC by 95% and 29% respectively. The results above suggested that the differential recognition of MHC I molecules of xeno-endothelial cells by human NK cells could be the major reason for higher NK cytotoxicity to PAEC. KIR might be the primary molecule that transduced inhibitory signals when endothelial cells were injured by NK cells.展开更多
Cellular immune response is a major barrier to xenotransplantation. Human tumor necrosis factor-α (hTNF-α) possesses cross-species activity and directly amplifies the immune rejection via the upregulation of adhesio...Cellular immune response is a major barrier to xenotransplantation. Human tumor necrosis factor-α (hTNF-α) possesses cross-species activity and directly amplifies the immune rejection via the upregulation of adhesion molecules on porcine endothelium. We investigated the role of protein tyrosine phosphorylation in the induction of expression of E-selectin and vascular cell adhesion molecule-1 (VCAM-1), and the augmentation of adhesion of human peripheral blood monocytes (PBMo) and natural killer cells (PBNK), after rhTNF-α-stimulation of porcine aortic endo-thelial cells (PAEC) in vitro. rhTNF-α-increased adhesiveness of PAEC for both PBMo and PBNK was dose-dependently reduced by pretreatment of PAEC with the selective protein tyrosine kinase (PTK) inhibitor genistein. The inhibitory effect occurred at the early time of PAEC activation triggered by rhTNF-α, and was completely reversible. PTK activity assay indicated that genistein also suppressed rhTNF-α stimulated activation of protein tyrosine展开更多
文摘The aim of the study was to investigate the effect of black soybean (BS) on the susceptibility of low-density lipoprotein (LDL) in hypercholesterolemic New Zealand white rabbits. Effects of the BS extract (BSE) and its components on monocyte adhesion of human aortic endothelial cells (HAECs), and adhesion molecule were investigated. Rabbits were divided into four groups, including control, 0.5% cholesterol with 20% casein (either with or without 0.5% vitamin E), and BS groups, all fed for 8 weeks. LDL was treated with 10 μM Cu2+ in vitro to determine the LDL lag time, and the vitamin E content of LDL was determined. The thickness of the tunica intima was measured on paraffin sections of thoracic aortas and aortic arches stained with Movat’s pentachrome. HAECs were pretreated with 100 μg/ml of BSE, and 10 μM of genistein, daidzein, cyanidin, and aspirin for 18 h, followed by tumor necrosis factor (TNF)-α (2 ng/ml) for 6 h, after which U937 cell adhesion was determined. Adhesion molecule expression was examined using ELISAs. The LDL lag time in the BS group was similar to that in the vitamin E group, while its lag time was significantly longer than those in the control and casein groups. The ratio of the intimal area/medial area of the aortic arch of the casein group was significantly higher than those in the control, BS, and vitamin E groups. The vitamin E group had the lowest value, and was closest to the control group. The BS group exhibited a significantly decreased atheroma region in the aortic arch compared to the casein group. Pre-incubation with BSE, genistein, daidzein, cyanidin, and aspirin significantly decreased adhesion by U937 monocytic cells to TNF-α stimulated HAECs. Genistein, daidzein, cyanidin, and aspirin significantly suppressed the expression of vascular cell adhesion molecule (VCAM)-1. Only genistein and aspirin significantly decreased intracellular adhesion molecule (ICAM)-1 expression compared to TNF-α treatment, while no treatments had any effect on E-selectin expression. BS significantly prolonged the LDL lag time and decreased the atheroma region of the aortic arch in hypercholesterolemic rabbits, thereby exerting an antiatherosclerotic effect. Presumably, the BSE downregulate intracellular redox-dependent signaling pathways in HAECs upon TNF-α stimulation through regulating NF-κB, thereby attenuating the inflammatory response in atherosclerosis. The antiatherogenic and anti-inflammatory effects of BS can be used as a nutraceutical for atherogenesis prevention.
基金Supported by the National Natural Science Foundation of China(81274031)
文摘[Objectives] To study the anti-inflammatory activity and mechanism of salvianolic acid A(Sal A), salvianolic acid C(Sal C), and rosmarinic acid(RA) on HAECs induced by TNF-α. [Methods] VE was used as a positive control and TNF-α induced human aortic endothelial cells(HAECs) were selected as the inflammation model cells. The levels of IL-6, MCP-1 were determined using ELISA. The mRNA levels of IL-6, MCP-1, ICAM-1, P-selectin and NF-κB were analyzed by quantitative RT-PCR. The protein expression of NF-κB was determined by western blot. The adhesion of U937 to HAECs was assessed by BCECF/AM labeling assay. Immunohistochemistry detection was used for ICAM-1 and P-selectin expression levels in TNF-induced HAECs, and DHE detection for the ROS production in HAECs induced by TNF-α. [Results] TNF-α-induced over-expressions of IL-6, MCP-1, NF-κB, ICAM-1 and P-selectin in HAECs were down-regulated by Sal A, Sal C and RA both in mRNA or protein levels, respectively. Meanwhile Sal A, Sal C and RA could significantly inhibit the production of ROS and U937 cells adhesion to HAECs. [Conclusions] Phenolic acids could inhibit TNF-α-induced inflammatory responses on HAECs and the mechanism might be related to inhibition of the production of ROS and blocking NF-κB signaling pathway.
文摘Objective To investigate the expression of endoplasmic reticulum stress(ESR)marker C/EBP homologous protein-10(CHOP-10)in the human aortic endothelial cells(HAEC)under the ischemia and hypoxia stress and to study the effects of atorvastatin on the process.Methods The cultured HAEC were divided into normal control group,ischemia/hypoxia model group。
文摘Cellular immune response is a major barrier to xenotransplantation, and cell adhesion is the first step in intercellular recognition. Flow-cytometric adhesion assay has been used to investigate the differential adhesions of monocyte (Mo), natural killer celi (NK) and T lymphocyte (T) present within human peripheral blood mononuclear cells (PBMC) to porcine aortic endothelial cells (PAEC), and to demonstrate the effect of human interferon-γ (hIFN-γ) or/and tumor necrosis factor-α (hTNF-α) pretreatment of PAEC on their adhesiveness f or different PBMC subsets. The preferential sequence for PBMC subset binding to resting PAEC is Mo, NK and T cells, among which T cells show the slightest adherence; hTNF- a can act across the species, and augment Mo, NK and T cell adhesion ratios by 40%, 110% and 3 times, respectively. These results confirm at the cell level that host Mo and NK cells are major participants in the cellular xenograft rejection, thereby, providing a prerequisite for further studying the
文摘Using human umbilical vein endothelial cells (HUVEC) and porcine aortic endothelial cells (PAEC) as target cells, human peripheral blood NK cells (PBNK) and NK92 cells as effector cells, the differential cytotoxicities of NK cells to allo- and xeno-endothelial cells were studied. The influence of MHC class I molecules on the cytotoxicity of human NK cells was assayed using acid treatment, and blockades of MHC class I antigens, CD94 and KIR (NKB1). The results indicated that the killing of PAEC by the two kinds of NK cells is higher than that of HUVEC. After acid-treatment, the cytotoxicity of the two kinds of NK cells to PAEC and HUVEC is significantly enhanced, but the magnitude of the enhancement is different. The enhancement of NK killing to acid treated HUVEC is much greater than that to PAEC. Blockade of CD94 mAb did not alter the NK cytotoxicity, while blockade of NKB1 mAb enhanced the cytotoxicity of PBNK to HUVEC and PAEC by 95% and 29% respectively. The results above suggested that the differential recognition of MHC I molecules of xeno-endothelial cells by human NK cells could be the major reason for higher NK cytotoxicity to PAEC. KIR might be the primary molecule that transduced inhibitory signals when endothelial cells were injured by NK cells.
基金This work was supported by the National Natural Science Foundation of China (Grant No. 39770700).
文摘Cellular immune response is a major barrier to xenotransplantation. Human tumor necrosis factor-α (hTNF-α) possesses cross-species activity and directly amplifies the immune rejection via the upregulation of adhesion molecules on porcine endothelium. We investigated the role of protein tyrosine phosphorylation in the induction of expression of E-selectin and vascular cell adhesion molecule-1 (VCAM-1), and the augmentation of adhesion of human peripheral blood monocytes (PBMo) and natural killer cells (PBNK), after rhTNF-α-stimulation of porcine aortic endo-thelial cells (PAEC) in vitro. rhTNF-α-increased adhesiveness of PAEC for both PBMo and PBNK was dose-dependently reduced by pretreatment of PAEC with the selective protein tyrosine kinase (PTK) inhibitor genistein. The inhibitory effect occurred at the early time of PAEC activation triggered by rhTNF-α, and was completely reversible. PTK activity assay indicated that genistein also suppressed rhTNF-α stimulated activation of protein tyrosine
