RESUMEN
The loss of endothelial connective integrity and endothelial barrier dysfunction can lead to increased vascular injury, which is related to the activation of endothelial inflammasomes. There are evidences that low concentrations of aspirin can effectively prevent cardiovascular diseases. We hypothesized that low-dose aspirin could ameliorate endothelial injury by inhibiting the activation of NLRP3 inflammasomes and ultimately prevent cardiovascular diseases. Microvascular endothelial cells were stimulated by lipopolysaccharide (2 μg/mL) and administrated by 0.1-2 mmol/L aspirin. The wild type mice were stimulated with LPS (100 μg/kg/day), and 1 h later treated with aspirin (12.5, 62.5, or 125 mg/kg/day) and dexamethasone (0.0182 mg/kg/day) for 7 days. Plasma and heart were harvested for measurement of ELISA and immunofluorescence analyses. We found that aspirin could inhibit NLRP3 inflammasome formation and activation in dose-dependent manner and has correlation between the NLRP3 inflammasome and the ROS/TXNIP pathway. We also found that low-concentration aspirin could inhibit the formation and activation of NLRP3 inflammasome and restore the expression of the endothelial tight junction protein zonula occludens-1/2 (ZO1/2). We assume that aspirin can ameliorate the endothelial layer dysfunction by suppressing the activation of NLRP3 inflammasome.
RESUMEN
Purpose: Excessive ultraviolet B (UVB) exposure causing corneal endothelium injury, including apoptosis, is a serious condition. Therefore, drugs that can inhibit apoptosis in corneal endothelial cells represent an effective strategy. Simvastatin is widely used as a specific inhibitor of 3-hydroxy-3-methyl-glutaryl-CoA reductase, can reduce levels of low density lipoprotein (LDL) cholesterol, and exerts anti-inflammatory effects. However, the protective effect of simvastatin on corneal endothelial cells remains unclear. Therefore, the aim of this study was to elucidate whether UVB promotes the initiation of apoptosis in corneal endothelial cells and injury reversible by simvastatin treatment. Methods: We detected the cell viability, subG1 population, and caspase-3 activity. Results: Results showed that simvastatin alleviates UVB-induced cell death, cell apoptosis, and caspase-3 activity. Conclusion: Our findings indicated that simvastatin alleviated UVB-induced corneal endothelial cell apoptosis via caspase-3 activity.
RESUMEN
Objective To investigate the effect of low pH on acid-sensing ion channel 1a( ASIC1a) expression in vascular endothelial cells induced by serum IgA 1 from Henoch-Sch?nlein purpura ( HSP ) children and regulatory role of ASIC1a in it.Methods Human dermal microvascular endothelial cells ( HDMECs) treated by serum IgA1 from children with HSP were incubated in different pH medium .ASIC1a, destrin and α-SM mRNA expressions in HDMECs were evaluated by real-time quantitative polymerase chain reaction (q-PCR).The level of inflammatory cytokines released by vascular endothelial cells was detected by ELISA .Moreover destrin and α-SM protein expres-sion in HDMECs was evaluated by Western blot .Results The results showed that in low pH condition , IgA1 from HSP children could induce the upregulation of ASIC 1a mRNA expression , stimulate IL-8, NO and TM release of vascular endothelial cells of HSP children .And blockers of ASICs could reduce acid-induced cytokine release of vascular endothelial cells .Moreover destrin and α-SM mRNA and protein expression in HDMECs of HSP children significantly decreased when exposure to extracellular acidosis .However blockers of ASICs increased destrin and α-SM mRNA and protein expression in vascular endothelial cells of HSP children .Conclusions These findings showed that activation of ASIC 1a could be involved in the vascular endothelial cell injury of HSP children .Blocking ASIC1a may have a significant protective effect on the inflammatory injury of vascular endothelial cells .
RESUMEN
Objective To investigate the role of mitochondrial respiratory chain (MRC) in induction of reactive oxygen species (ROS) by hyperbaric oxygen (HBO).Methods Superoxide anion (O·-2) specific fluorescence probe DHE and mitochondrial O·-2 probe Mito-SOX were used to label ROS in human umbilical vein endothelium cells(HUVEC) microscopically after HBO exposure.Results After HBO exposure, O·-2 increased (31±8)% and (137±19) % in whole cells and in mitochondria, respectively (P<0.01).These increments were suppressed by MRC complex Ⅱ inhibitor TTFA for (55±11)% in whole cells (P<0.05)and (61±8) % in mitochondria(P<0.01).Conclusion MRC may be the main source of ROS induced by HBO in HUVECs.
RESUMEN
Aim To investigate the role of captopril in insulin resistance of endothelial cells induced by high glucose.Methods 1 .Improvement effect of captopril on insulin resistance in HUVECs was observed.The HUVECs were seeded in a 6-well plate and were ran-domly divided into 5 groups,namely,control group, IR group,IR together with different Cap concentrations (low,medium and high concentration),respectively. 2.Improvement effect of Cap on insulin resistance was mediated by PPARγin HUVECs.HUVECs were ran-domly divided into 6 groups,namely,control group, control +PPARγinhibitor (PI)(1 .0 μmol · L -1 ) group,IR group,IR +PI(1 .0 μmol·L -1 )group,IR +Cap(1 ×1 0 -5 mol·L -1 ) group,and IR +Cap +PI (1 .0 μmol·L -1 )group.All indicators were detected. Results After HUVECs were incubated with media containing 33 mmol·L -1 of glucose for 48 h,the NO levels were significantly decreased while ET-1 levels were significantly elevated,showing a significant differ-ence between IR group and control group (P 0.05).When the HUVECs in IR group were treated with DMEM containing glucose (33 mmol·L -1 )for 48 h and insulin for 30 min,the expression levels of PPARγmRNA and its protein in Cap groups were simi-lar to those in the IR group,and there was no signifi-cant difference between the two groups (P >0.05 );however, the expression levels of phosphorylated PPARγprotein in Cap groups were increased compared with IR group (P <0.05).The levels of NO were sig-nificantly increased whereas the levels of ET-1 were decreased in Cap groups,which had significant differ-ences compared with IR group (P <0.05).Nonethe-less,pre-treating with GW9662,a PPARγinhibitor, the improvement effects of Cap were markedly abol-ished.Conclusions Captopril could improve high glucose-induced insulin resistance of endothelial cells mediated by PPARγ,and the underlying mechanisms are related to the activation of PPARγ,rather than its expression.
RESUMEN
Objective To study the effects of Exendin-4 (Ex-4) on the function of endothelial cells by investigating the relationship of Ex-4 induced nitric oxide (NO) release from pig iliac endothelial cells (PIEC) and changes in intracellular calcium. Methods Glucagon-like peptide-1 receptor (GLP-1R) was confirmed to be present in PIEC by immunofluorescence assay. Intracellular NO production was probed using DAF-FM DA, and then measured with fluorescence microplate reader at the timepoints of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12h. The intracellular Ca2+ concentration was probed with Fura-2 AM at 0-30min, 4-4.5h and 8-8.5h and measured with fluorescence microplate reader. The relationship between Ex-4 induced NO release in PIEC and intracellular Ca2+ concentration was evaluated. Results GLP-1R was expressed in PIEC. Ex-4 increased the intracellular NO level in PIEC in a time-dependent manner. NO level was significantly higher 4 hours after Ex-4 treatment compared with that of 0-hour group (P<0.05). The intracellular NO level reached the top value 8 hours after Ex-4 treatment (P<0.05). The intracellular Ca2+ concentrations were significantly higher during the periods of 0-30min, 4-4.5h and 8-8.5h after Ex-4 treatment than those of their control groups (2.042±2.115 vs 0.634±0.352, P<0.01; 0.413±0.154 vs 0.113±0.111, P<0.01; 0.309±0.133 vs 0.063±0.120, P<0.01). During the period of 0-30min after Ex-4 treatment, the intracellular Ca2+ concentration increased gradually (fitted curve: y=0.106x+1.326); while during the periods of 4-4.5h (y=0.003x+0.374) and 8-8.5h (y=-0.001x+0.324) after Ex-4 treatment, the intracellular Ca2+ concentrations, and the NO levels were higher than those of their control groups, but without significant variation. Conclusions Ex-4 may increase the NO release in endothelial cells, and intracellular calcium ions may participate in the process.
RESUMEN
Objective To study the effects of Exendin-4 (Ex-4) on the function of endothelial cells by investigating the relationship of Ex-4 induced nitric oxide (NO) release from pig iliac endothelial cells (PIEC) and changes in intracellular calcium. Methods Glucagon-like peptide-1 receptor (GLP-1R) was confirmed to be present in PIEC by immunofluorescence assay. Intracellular NO production was probed using DAF-FM DA, and then measured with fluorescence microplate reader at the timepoints of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12h. The intracellular Ca2+ concentration was probed with Fura-2 AM at 0-30min, 4-4.5h and 8-8.5h and measured with fluorescence microplate reader. The relationship between Ex-4 induced NO release in PIEC and intracellular Ca2+ concentration was evaluated. Results GLP-1R was expressed in PIEC. Ex-4 increased the intracellular NO level in PIEC in a time-dependent manner. NO level was significantly higher 4 hours after Ex-4 treatment compared with that of 0-hour group (P<0.05). The intracellular NO level reached the top value 8 hours after Ex-4 treatment (P<0.05). The intracellular Ca2+ concentrations were significantly higher during the periods of 0-30min, 4-4.5h and 8-8.5h after Ex-4 treatment than those of their control groups (2.042±2.115 vs 0.634±0.352, P<0.01; 0.413±0.154 vs 0.113±0.111, P<0.01; 0.309±0.133 vs 0.063±0.120, P<0.01). During the period of 0-30min after Ex-4 treatment, the intracellular Ca2+ concentration increased gradually (fitted curve: y=0.106x+1.326); while during the periods of 4-4.5h (y=0.003x+0.374) and 8-8.5h (y=-0.001x+0.324) after Ex-4 treatment, the intracellular Ca2+ concentrations, and the NO levels were higher than those of their control groups, but without significant variation. Conclusions Ex-4 may increase the NO release in endothelial cells, and intracellular calcium ions may participate in the process.
RESUMEN
@#Objective To investigate the morphological effect of fluid shear stress on pig iliac endothelium cells cultured solely or co-cultured with pig small intestinal submucosa.Methods The shear stress of 40×10-5 N/cm2 were carried out for 12 h on both groups.The images were recorded every 30 min.The directional angles were calculated.Results In the group of cell cultured solely:The defluvium of cells was obvious at the 1st hour,but the shape of cells didn't change.At the 4th hour,the defluvium of cells was little,the cell became round from its initiatory polygon shape.At the 8th hour,the defluvium of cell could not be observed.The shape of cells became fusiform and gracile.The cells arranged along the direction of flow field in the local area.At the 12th hour,the cells became more and more gracile.The trend of realignment of cells along the direction of flow field was obviously.The directional angles of cells at the 12th hour was significantly different from the zero hour.In the group of cell co-cultured with small intestinal submucosa:At the 1st hour,some of cells were brushed off mildly.The defluvium of cells could not been observed since the 2nd hour.The directional angles didn't change significantly in the 12 hours.Conclusion The shear stress of 40×10-55 N/cm2 cannot influence the cell of co-cultured but do influence the cell cultured solely.
RESUMEN
Objective To investigate the effects of ovariectomy and cholesterol rich diet on the change of estrogen receptor content of vascular endothelial cells and hearts of female rats. Methods The receptor binding assay (RBA) was adopted to measure the content of estrogen receptors, and the serum levels of estradiol and lipids were measured. Results The content of ER was much lower in hearts and VEC of ovariectomy〔(0.51?0.09) fmol/mg pro,(6.73 ? 0.52) fmol/106 cell〕and cholesterol rich rats〔(0.97?0.12) fmol/mg pro,(9.15 ?0.53) fmol/106 cell〕than pseudo-operation rats 〔(2.08?0.15) fmol/mg pro ,(17.66?1.26) fmol/106 cell , P
RESUMEN
Objective To investigate the effect of pronuciferine on apoptosis of cultured human umbilical vein endothelium cells(HUVECs) induced by angiotensin Ⅱ(AngⅡ).Methods HUVECs cell line ECV304 was cultured in vitro,pretreated with Captopril(10 ?mol/L) or pronuciferine 10,1,0.1,0.01 ?mol/L for 30 min,respectively,then treated with AngⅡ(1 ?mol/L).Cell-morphosis was observed by light microscope.Cells viability was assessed by MTT assay.Production of nitric oxide(NO),activities of total nitric oxide synthase(tNOS),and inducible nitric oxide synthase(iNOS) were measured by colorimetry.Apoptosis rate was measured by Flow Cytometer(FCM).Results AngⅡ induced typical endothelial cell apoptosis and the apoptosis rates were significantly higher than those of the control group((P
RESUMEN
Adhesion of lymphocytes to EC is one of the important steps of lympho. cyte migration into the tissues outside vessels; and the components of blood plasma may have some effect on the adhesion of lymphocytes. In our experiments, we observed that PNIP, normally present in blood and being able to inhibit the proliferation of lymphocytes stimulated by antigens or mitogens, could significantly inhibit the adhesion of the lymphocytes from blood, thymus and mesenteric lymph nodes (MLN) to the aortic EC, using the aortic EC monolayer preparation (Hautchen technique). Meanwhile, the indirect immunofluorescent test also showed us that PNIP might result in a remarkable decrease in percentage of SmIgG positive B lymphocytes in the total lymphocytes adhering to the aortic EC. These results imply that PNIP may suppress the adhesion of lymphocytes or other leukocytes to EC in the development of inflammation, atherosclerosis or thrombosis.