Background Inhibition of aging of vascular endothelial cells （VECs） may delay aging and prolong life. The goal of this study was to prepare anti-CD31 monoclonal antibody conjugated PEG-modified liposomes containing ...Background Inhibition of aging of vascular endothelial cells （VECs） may delay aging and prolong life. The goal of this study was to prepare anti-CD31 monoclonal antibody conjugated PEG-modified liposomes containing the AU-rich region connecting factor 1 （AUF1） gene （CD31-PILs-AUF1） and to explore the effects of targeting CD31-PILs-AUF1 to aging VECs. Methods The mean particle sizes of various PEGylated immunoliposomes （PILs） were measured using a Zetasizer Nano ZS. Gel retardation assay was used to confirm whether PILs had encapsulated the AUF1 plasmid successfully. Fluorescence microscopy and flow cytometry were used to quantify binding of CD31-PILs-AUF1 to target cells. Flow cytometry was also used to analyze the cell cycles of aging bEnd3 cells treated with CD31-PILs-AUF1. We also developed an aging mouse model by treating mice with D-galactose. Enzyme-linked immunosorbent assay （ELISA） was used to evaluate the levels of interleuldn-6 （IL-6） and tumor necrosis factor-or （TNF-ct）. The malondialdehyde （MDA） and the superoxide dismutase （SOD） levels were detected by commercial kits. Hematoxylin-eosin （HE） staining was used to determine whether treatment with CD31-PILs-AUF 1 was toxic to the mice. Results CD31-PILs-AUF 1 specifically could targeted bEnd3 VECs and increased the percentage of cells in the S and G2/M phases of aging bEnd3 cells. ELISA showed that content of the IL-6 and TNF-ct decreased in CD31-PILs-AUF1 group. The level of SOD increased, whereas MDA decreased in the CD31-PILs-AUF1 group. Additionally, CD31-PILs-AUF 1 was not toxic to the mice. Conclusion CD31-PILs-AUF 1 targets VECs and may delay their senescence.展开更多
Exogenous electrical stimulation plays an important role on endothelial cells and vessels. Study on the effect of electrical stimulation on endothelial cells might be helpful to regulate cell metabolism on a certain e...Exogenous electrical stimulation plays an important role on endothelial cells and vessels. Study on the effect of electrical stimulation on endothelial cells might be helpful to regulate cell metabolism on a certain extent, or provide new ideas for vascular tissue engineering. A bioreactor that can apply electrical stimulation was designed on the basis of parallel-plate chamber. To investigate the possible effect of electrical stimulation on human umbilical vein endothelial cells （HUVECs）, the different levels and duration＇s electrical stimulations were applied to HUVECs in culture. The cell morphology was observed by microscopy and the levels of endothelial nitric oxide synthase （eNOS） gene were measured by RT-PCR. Different levels and durations of electrical stimulation produce different effects on eNOS gene expression. The eNOS gene expressions of the experimental group ceils under the voltage of 50 mV, 100 mV, 150 mV and 200 mV were significantly lower than that of the control group after 3-hour electrical stimulation, while the eNOS gene levels of the experimental group cells in 6-hour electrical stimulation were higher than those of the control group under all tested voltages. After 12-hour stimulation, the eNOS gene levels of HUVECs decreased under 50 mV, and then gradually increased until 200 inV. The low voltage of 6-hour electrical stimulation is more appropriate for HUVECs growth.展开更多
In order to investigate whether lipoxin A4 （LXA4） has an antagonistic effect on lipopolysaccharide （LPS）-induced synthesis of interleukin （IL）-β3, IL-6 and IL-8 in rat pulmonary microvascular endothelial cells ...In order to investigate whether lipoxin A4 （LXA4） has an antagonistic effect on lipopolysaccharide （LPS）-induced synthesis of interleukin （IL）-β3, IL-6 and IL-8 in rat pulmonary microvascular endothelial cells （PMVEC）, and to explore the molecular mechanisms of signal pathway in LXA4 actions, cultured PMVEC were treated with LPS, with or without preincubation with LXA4. Proteins of IL-β3, IL-6 and IL-8 in supernatant were analyzed by enzyme-linked immunosorbent assay （ELISA）. Expressions of mRNA of IL-β3, IL-6 and IL-8 were determined by RT-PCR. Expressions of phosphorylation of phosphoinositide 3-kinase （PI3-K） and myeloid differentiation factor 88 （MyD88） were analyzed by Western blot. Activities of DNA-binding of nuclear factor-kappa B （NF-κB） and activator protein-1 （AP-1） were measured by electrophoretic mobility shift assay （EMSA）. The results showed that LPS induced production of IL-β3, IL-6 and IL-8 in rat PMVEC via MyD88/PI3-K/NF-κB and AP-1 pathway-dependent signal transduction. LPS-stimulated expression of PI3-K, activities of NF-κB and AP-1, secretion of protein and expression of mRNA of IL-β3, IL-6 and IL-8 but not MyD88 expression in PMVEC were inhibited by LXA4 in a dose-dependent manner. In conclusion, LXA4 inhibits synthesis of IL-β3, IL-6 and IL-8 by down-regulation of PI3-K/NF-κB and AP-1 signal pathway in PMVEC.展开更多
基金This work was supported by grants from the Guangxi Natural Science Foundation （No. 2015GXNSFAA139217 and 2016GXNSFAA380231）, a grant from The Scientific Research Fund of Guangxi Education Department （No. YB2014057） entitled ＂AU-rich region connecting factor 1 targeted vascular endothelial cells for anti-aging＂, a grant from the Youth Foundation in Guangxi Medical Univer- sity （No. GXMUYSF201328）, a grant from the Undergraduate Innovative plan in Guangxi （No. 201510598012）, and a grant from the Guangxi Education Department Grant entitled ＂Innovation Project of Guangxi Graduate Educa- tion＂.
文摘Background Inhibition of aging of vascular endothelial cells （VECs） may delay aging and prolong life. The goal of this study was to prepare anti-CD31 monoclonal antibody conjugated PEG-modified liposomes containing the AU-rich region connecting factor 1 （AUF1） gene （CD31-PILs-AUF1） and to explore the effects of targeting CD31-PILs-AUF1 to aging VECs. Methods The mean particle sizes of various PEGylated immunoliposomes （PILs） were measured using a Zetasizer Nano ZS. Gel retardation assay was used to confirm whether PILs had encapsulated the AUF1 plasmid successfully. Fluorescence microscopy and flow cytometry were used to quantify binding of CD31-PILs-AUF1 to target cells. Flow cytometry was also used to analyze the cell cycles of aging bEnd3 cells treated with CD31-PILs-AUF1. We also developed an aging mouse model by treating mice with D-galactose. Enzyme-linked immunosorbent assay （ELISA） was used to evaluate the levels of interleuldn-6 （IL-6） and tumor necrosis factor-or （TNF-ct）. The malondialdehyde （MDA） and the superoxide dismutase （SOD） levels were detected by commercial kits. Hematoxylin-eosin （HE） staining was used to determine whether treatment with CD31-PILs-AUF 1 was toxic to the mice. Results CD31-PILs-AUF 1 specifically could targeted bEnd3 VECs and increased the percentage of cells in the S and G2/M phases of aging bEnd3 cells. ELISA showed that content of the IL-6 and TNF-ct decreased in CD31-PILs-AUF1 group. The level of SOD increased, whereas MDA decreased in the CD31-PILs-AUF1 group. Additionally, CD31-PILs-AUF 1 was not toxic to the mice. Conclusion CD31-PILs-AUF 1 targets VECs and may delay their senescence.
文摘Exogenous electrical stimulation plays an important role on endothelial cells and vessels. Study on the effect of electrical stimulation on endothelial cells might be helpful to regulate cell metabolism on a certain extent, or provide new ideas for vascular tissue engineering. A bioreactor that can apply electrical stimulation was designed on the basis of parallel-plate chamber. To investigate the possible effect of electrical stimulation on human umbilical vein endothelial cells （HUVECs）, the different levels and duration＇s electrical stimulations were applied to HUVECs in culture. The cell morphology was observed by microscopy and the levels of endothelial nitric oxide synthase （eNOS） gene were measured by RT-PCR. Different levels and durations of electrical stimulation produce different effects on eNOS gene expression. The eNOS gene expressions of the experimental group ceils under the voltage of 50 mV, 100 mV, 150 mV and 200 mV were significantly lower than that of the control group after 3-hour electrical stimulation, while the eNOS gene levels of the experimental group cells in 6-hour electrical stimulation were higher than those of the control group under all tested voltages. After 12-hour stimulation, the eNOS gene levels of HUVECs decreased under 50 mV, and then gradually increased until 200 inV. The low voltage of 6-hour electrical stimulation is more appropriate for HUVECs growth.
文摘In order to investigate whether lipoxin A4 （LXA4） has an antagonistic effect on lipopolysaccharide （LPS）-induced synthesis of interleukin （IL）-β3, IL-6 and IL-8 in rat pulmonary microvascular endothelial cells （PMVEC）, and to explore the molecular mechanisms of signal pathway in LXA4 actions, cultured PMVEC were treated with LPS, with or without preincubation with LXA4. Proteins of IL-β3, IL-6 and IL-8 in supernatant were analyzed by enzyme-linked immunosorbent assay （ELISA）. Expressions of mRNA of IL-β3, IL-6 and IL-8 were determined by RT-PCR. Expressions of phosphorylation of phosphoinositide 3-kinase （PI3-K） and myeloid differentiation factor 88 （MyD88） were analyzed by Western blot. Activities of DNA-binding of nuclear factor-kappa B （NF-κB） and activator protein-1 （AP-1） were measured by electrophoretic mobility shift assay （EMSA）. The results showed that LPS induced production of IL-β3, IL-6 and IL-8 in rat PMVEC via MyD88/PI3-K/NF-κB and AP-1 pathway-dependent signal transduction. LPS-stimulated expression of PI3-K, activities of NF-κB and AP-1, secretion of protein and expression of mRNA of IL-β3, IL-6 and IL-8 but not MyD88 expression in PMVEC were inhibited by LXA4 in a dose-dependent manner. In conclusion, LXA4 inhibits synthesis of IL-β3, IL-6 and IL-8 by down-regulation of PI3-K/NF-κB and AP-1 signal pathway in PMVEC.
