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Pulmonary endothelial barrier dysfunction is a hallmark of clinical pulmonary edema and contributes to the development of acute lung injury (ALI). Here we reported that ruscogenin (RUS), an effective steroidal sapogenin of Radix Ophiopogon japonicus, attenuated lipopolysaccharides (LPS)-induced pulmonary endothelial barrier disruption through mediating non-muscle myosin heavy chain IIA (NMMHC IIA)‒Toll-like receptor 4 (TLR4) interactions. By in vivo and in vitro experiments, we observed that RUS administration significantly ameliorated LPS-triggered pulmonary endothelial barrier dysfunction and ALI. Moreover, we identified that RUS directly targeted NMMHC IIA on its N-terminal and head domain by serial affinity chromatography, molecular docking, biolayer interferometry, and microscale thermophoresis analyses. Downregulation of endothelial NMMHC IIA expression in vivo and in vitro abolished the protective effect of RUS. It was also observed that NMMHC IIA was dissociated from TLR4 and then activating TLR4 downstream Src/vascular endothelial cadherin (VE-cadherin) signaling in pulmonary vascular endothelial cells after LPS treatment, which could be restored by RUS. Collectively, these findings provide pharmacological evidence showing that RUS attenuates LPS-induced pulmonary endothelial barrier dysfunction by inhibiting TLR4/Src/VE-cadherin pathway through targeting NMMHC IIA and mediating NMMHC IIA‒TLR4 interactions.
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Disruption of pulmonary endothelial permeability and associated barrier integrity increase the severity of acute respiratory distress syndrome (ARDS). This study investigated the potential ability of the human immunodeficiency virus-1 (HIV-1) integrase inhibitor raltegravir to protect against acute lung injury (ALI) and the underlying mechanisms. Accordingly, the impact of raltegravir treatment on an in vitro lipopolysaccharide (LPS)-stimulated human pulmonary microvascular endothelial cell (HPMEC) model of ALI and an in vivo LPS-induced two-hit ALI rat model was examined. In the rat model system, raltegravir treatment alleviated ALI-associated histopathological changes, reduced microvascular permeability, decreased Evans blue dye extravasation, suppressed the expression of inflammatory proteins including HMGB1, TLR4, p-NF-κB, NLRP3, and MPO, and promoted the upregulation of protective proteins including claudin 18.1, VE-cadherin, and aquaporin 5 as measured via western blotting. Immunohistochemical staining further confirmed the ability of raltegravir treatment to reverse LPS-induced pulmonary changes in NLRP3, claudin 18.1, and aquaporin 5 expression. Furthermore, in vitro analyses of HPMECs reaffirmed the ability of raltegravir to attenuate LPS-induced declines in VE-cadherin and claudin 18.1 expression while simultaneously inhibiting NLRP3 activation and reducing the expression of HMGB1, TLR4, and NF-kB, thus decreasing overall vascular permeability. Overall, our findings suggested that raltegravir may represent a viable approach to treating experimental ALI that functions by maintaining pulmonary microvascular integrity.
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Objective To investigate the prognostic significance of serum VE-cadherin in patients with septic shock. Methods A prospective observation study was performed between January 2016 and December 2017, forty-eight septic shock patients from intensive care unit (ICU) were enrolled, and 25 healthy volunteers served as the controls. Meanwhile, patients in the septic shock group were divided into two subgroups of the survival and death groups according to the 28-day mortality. The dynamic value changes of serum VE-cadherin (VE-Cad), vascular endothelial growth factor (VEGF), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were monitored on the 1st, 3rd and 7th day after admission. Results The serum VE-Cad level of the septic group was significantly higher than that of the control group on the 1st day (3.02±0.18 ng/mL vs. 0.26±0.05 ng/mL, t=3.275, P=0.002). There was a positive correlation between VE-Cad level and VEGF, TNF-α and IL-6 (r=0.826, 0.723, and 0.870, respectively; P<0.01). The PaO2/FiO2 and serum albumin (ALB) level of the death group were lower than that of the survival group, and extravascular lung water index (EVLWI), serum lactate, mechanical ventilation time, 7 day fluid balance, APACHE Ⅱ and SOFA levels of the death group were higher than those of the survival group. The serum VE-Cad levels of the death group were higher than those of the survival group on the 1st, 3rd and 7th day. The serum VE-Cad levels were positively correlated with APACHE Ⅱ and SOFA. The area under the ROC curve of VE-Cad predicting septic shock death on day 1 was 0.723 (95％CI 0.568-0.878). The sensitivity and specificity of VE-Cad with cut-off of 3.1 ng/mL in predicting septic shock death were 60％ and 70.83％, respectively. Conclusions Serum VE-Cad is positively correlated with disease severity and could predict a poor outcome in septic shock patients.
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Objective To investigate the prognostic significance of serum vascular endothelial cadherine (VE-Cad) in patients with acute respiratory distress syndrome (ARDS) induced by sepsis . Methods A prospective observational study was performed between June 2015 and Dec 2017 ,and 48 ARDS patients induced by sepsis from intensive care unit (ICU) were enrolled .And 30 healthy volunteers were enrolled as control .ARDS group was divided into mild group (n=17) ,moderate group (n=18) and severe group (n= 13) .The dynamic levels of serum VE-Cad ,tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were monitored at day 1 ,3 and 7 of admission .Clinical data including extravascular lung water index (EVLWI) ,pulmonary vascular permeability index (PVPI) ,lung injury score (LIS) , APACHEⅡand SOFA were also collected .The t-test or chi square test were used in the comparison between the two groups .One-way ANOVA was used for comparison among multiple groups .Results The serum VE-Cad level of septic group was higher than control group at day 1 of admission ([5 .67 ± 0 .29] vs [0 .28 ± 0 .03] μg/L ,t= 101 .2 , P< 0 .01) .The serum VE-Cad levels in the mild group , moderate group and severe group were (1 .52 ± 0 .59) ,(3 .45 ± 0 .68) ,and (4 .68 ± 0 .53) μg/L , respectively (F=15 .45 ,P<0 .01) .There were positive correlation between VE-Cad levels and EVLWI , PVPI ,LIS ,TNF-αand IL-6 (r=0 .640 ,0 .601 ,0 .507 ,0 .584 ,and 0 .456 ,respectively ,all P<0 .01) . The PaO2/FiO2 and serum albumin level in death group (n=17) were lower than survival group (n=31) ([146 .74 ± 16 .45] vs [245 .42 ± 12 .13] mmHg [1 mmHg=0 .133 kPa] ,t=23 .72 ,P<0 .01 ;[23 .18 ± 3.24]vs[29.16±3.45]g/L,t=5.865,P< 0.01,respectively),andEVLWI ,PVPI,LIS,serum lactate ,mechanical ventilation time ,7 d fluid balance ,APACHEⅡ and SOFA in death group were all higher than survival group .The serum VE-Cad levels at day 1 ,3 and 7 in death group were all higher than survival group ([4 .72 ± 0 .96] vs [3 .36 ± 0 .47]μg/L ,t=8 .801;[3 .87 ± 0 .28] vs [1 .95 ± 0 .42]μg/L , t=16 .86 ;[3 .92 ± 0 .53] vs [0 .96 ± 0 .28]μg/L ,t=25 .42 ,respectively ,all P<0 .01) .The area under curve (AUC ) of VE-Cad for ARDS outcome prediction was 0 .878 with sensitivity of 100 .00％ and specificity of 58 .06％ with a cutoff of 3 .035 μg/L .Conclusion Serum VE-Cad level increases in patients with ARDS induced by sepsis ,and positively correlates with disease severity ,which could be a potential predictor for prognosis .
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Objective:To investigate the influence and related mechanisms of T7 peptide derived from tumstatin on angiogenesis in vitro.Methods:HUVECs were incubated in hypoxia chamber(37℃,1％ O2,5％ CO2,94％ N2) to simulate the hypoxic microenvironment in tumors,and grouped into:hypoxia group,hypoxia+T7 peptide (1.0 μ mol/L) group,and hypoxia+T7 peptide(2.0μ mol/L) group.The tube formation assay was applied to analyze the influence of T7 peptide on angiogenesis under hypoxia.Cell Counting Kit-8 was applied to observe the cell viability.The apoptosis rate was detected with Annexin V-FITC,and the expression levels of pro-apoptotic protein BAX and anti-apoptotic protein Bcl-2 were observed with western blot.Immunofluorescence and western blot were used to detect the expression of VE-cadherin.Results:Under hypoxic condition,T7 peptide significantly inhibited capillary-like tube formation (P＜0.05),inhibited ECs viability(P＜0.05),and increased the ECs apoptosis rate in vitro;T7 peptide resulted in the downregulation of anti-apoptotic protein Bcl-2 (P＜0.01)and upregulation of pro-apoptotic protein BAX(P＜0.05);the expression of VE-cadherin was downregulated by T7 peptide significantly(P＜0.05).Conclusions:T7 peptide could execute its anti-angiogenic activity via inhibition of ECs viability,induction of ECs apoptosis rate,and downregulation of VE-cadherin expression.
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Objective To monitor the association between the expression of ve-cadherin and mesentery microcirculation blood flow of sepsis CASP rats,Analysis the relationship of each other.Methods Based on the standard criteria,we established sepsis CASP rats,and the rats were divided into several groups according to the gauge of the vein detained needle in the ascending colon,every group has six rats.The groups consisted of Sepsis A group 22 G(0.9 mm× 25 mm,33 ml/min),sepis B group 20 G(1.1 mm ×32 mm,54 ml/min),sepsis C group 18 G(1.3 mm ×32 mm,80 ml/min),sepsis D group 14 G(2.0 mm ×45 mm,270 ml/min),in addition we established the control group.Real-time observation was performed by microscope on the 6 h's blood velcocity of each sepsis CASP rats group.Then we applied the immunohistochemistry to quantitatively analyze the expression of ve-cadherin of each group.Results The blood velcocity of the mesentery microcirculation in control group was (583.21 ±52.39) μm/s,it was higher compared with the sepsis D group(213.30 ±52.39) μm/s (P ＜0.05),the blood velocity in sepsis A group was (482.71 ± 58.62) μm/s,higher compared with the sepsis D group(P ＜ 0.05).The score of the immunohistochemistry quantitative analysis of mesentery ve-cadherin in control group was 11.17 ±0.34,higher compared with the sepsis D group(5.43 ±0.43)(P ＜0.01).The score of sepsis A group was 10.07 ±0.30,higher compared with the sepsis D group(P＜0.05).Conclusions The expression of ve-cadherin of sepsis CASP rats had positive correlation with the blood velocity of the mesentery microcirculation of sepsis CASP rats,and it can indirectly reflect the degree of the sepsis.
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Objective To investigate the influence of Ulinastatin (UTI) on the hyper-permeability of vascular endothelial cells induced by tumor necrosis factor alpha (TNF-α).Methods Inflammation model was induced by TNF-α in human umbilical vein endothelial cell line (EA.hy926).The experiment was designed into 4 groups:normal group,TNF-α group,UTI group and TNF-α with UTI (U + T) group.Methyl thiazolyl tetrazolium (MTT) method and epithelial voltameter (EVOM) method were used to measure cell viability [absorbance (A) value] and transepithelial electrical resistance (TER) of EA.hy926 cells respectively.The expression of VE-cadherin was measured by reverse transcription polymerase chain reaction (RT-PCR) and immunocytochemistry.Results Compared with normal group,the TER of EA.hy926 cells induced by TNF-α was significantly decreased (67.200 ± 8.937 vs.33.600 ± 8.771,P =0.010).The permeability in EA.hy926 cells increased obviously.The hyper-permeability of EA.hy926 cells induced by TNF-α could be alleviated by UTI at the concentrations of 1-100 U/mL in a dose-dependent manner (40.133 ±7.484 vs.33.600 ±8.771,P=0.382;49.232 ± 3.162 vs.33.600 ± 8.771,P =0.044;63.700 ± 8.515 vs.33.600 ± 8.771,P =0.013).The expression of VE-cadherin mRNA reduced significantly in the TNF-α group (1.089 ±0.018 vs.0.835±0.021,P =0.000) compared with normal group.This effect of TNF-α could be attenuated by UTI.When EA.hy926 cells exposed to UTI at 10 U/mL and 100 U/mL,a significant increase of the expression of VE-cadherin mRNA was observed (0.976 ±0.014 vs.0.835 ±0.021,P =0.001;1.115 ±0.015 vs.0.835 ± 0.021,P =0.000).And the inhibition of UTI manifested a dose-dependent manner (1-100 U/mL).The results of the immunocytochemistry showed that the expression of VE-cadherin in TNF-o group was decreased significantly (0.061 ± 0.013 vs.0.093 ± 0.014,P =0.049) compared with normal group.And the low-expression of VE-cadherin could be alleviated by UTI (0.032 ± 0.004 vs.0.061 ± 0.013,P =0.016).Conclusion The high permeability of EA.hy926 cells induced by TNF-α could be inhibited by UTI at the concentrations of 1-100 U/mL in a dose-dependent manner.
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Objective:To investigate the inhibitory effect of Dickkopf-1 (Dkk1) on vasculogenic mimicry (VM) formation and the relevant mechanism. Methods:CD34-PAS dual staining and immunohistochemical staining were used to detect and analyze the re-lationship between VM existence and Dkk1 expression in 217 human colon cancer tissue samples;three dimensional (3D) culture was used to detect the influence of Dkk1 on tube structure formation and on VE-cadherin expression;a subcutaneous mouse xenograft mod-el was made to further validate the inhibitory role of Dkk1 on VM formation in vivo. Results:VM-positive samples indicated a lower expression of Dkk1(P<0.05);colon cancer cells with Dkk1 overexpression exhibited a decreased ability to form tube-like structure and a decreased expression of VE-cadherin;Dkk1 inhibited the VM-formation abilities of human colorectal carcinoma cell line xenograft tu-mor tissue. Conclusion:Dkk1 inhibits the VM formation of colon cancer.
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Objective: To clarify that protection of Tongxinluo against myocardial ischemia/reperfusion injury relates to enhancing endothelial barrier in diabetic rats. Methods: A total of 32 Zucker diabetic rats were randomized into 4 groups:Sham group, Model group, Insulin group and Tongxinluo group, n=8 in each group. In addition, there was a Control group containing 8 non-diabetic Zucker rats. Myocardial ischemia/reperfusion injury model was established by a 45-min ischemia and 3-h reperfusion protocol. The size of infarction was detected by pathological staining, the microvascular permeability was examined by Miles assay to obtain the lfuorescein isothiocyanate concentration in myocardial tissue, the Triton X-100 soluble and insoluble VE-cadherin was measured by membrane protein extraction and western blot analysis. Results:The size of infarction in Model group was obviously higher than that in Control group (55.2 ± 1.4)%vs (36.2± 1.3)%,P0.05. Both insulin group and Tongxinluo group got lower lfuorescein isothiocyanate concentration (all P Conclusion: Protection of Tongxinluo against myocardial ischemia/reperfusion injury in diabetic rats is as effective as insulin, but the effect is independent of reducing blood glucose and may be related to enhancing endothelial barrier.
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Objective To investigate the possible correlation between obstructive sleep apnea hypopnea syndrome(OSAHS)and serum epithelial neutrophil-activing peptide-78(ENA-78)and VE-cadherin in pregnant patients.Methods One hundred and two pregnant women combined with OSAHS,42 of which were with hypertension,and 36 healthy pregnant women were underwent polysomnography(PSG),and their apnea hypopnea index(AHI)was recorded.Based on the apnea hypopnea index(AHI),they were divided into mild OSAHS group(36 patients),moderate OSAHS group(34 patients)and severe OSAHS group(32 patients)Serum ENA-78 and VE-cadhefin levels were detected by enzyme-linked immunosorbent assay(ELISA),and mean arterial pressure(MAP)was measured simultaneously.Results The values of AHI,MAP,ENA-78 and VE-cadherin in OSAHS with hypertension were significantly higher than those of OSAHS only group(t =13.46,11.65,12.47,13.36,all P ＜ 0.01)and the control group(t =16.72,15.31,16.35,16.73,all P ＜ 0.01).The levels of serum ENA-78 and VE-cadherin were gradually significantly decreased in the order of severe OSAHS group,moderate OSAHS group and mild OSAHS group(P ＜0.01).The AHI of OSAHS group was significantly correlated with MAP,VE-cadherin,ENA-78(r =0.75,0.71,0.68,all P ＜ 0.01).ENA-78 of OSAHS group were positively correlated with MAP and VE-cadherin(r =0.64,0.67,all P ＜ 0.01).Conclusions The condition OSAHS in pregnant patients is closely related to the levels of ENA-78 and VE-cadherin.Pregnancy combined with OSAHS may take part in the pathological process of hypertensive disorder complicating pregnancy.
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Objective To investigate the changes of high mobility group box-1 ( HMGB1 ) and VE-cadherin in serum of children with viral myocarditis and their clinical significance. Methods The serum levels of HMGB1 and VE-cadherin were detected by ELISA in 52 children with viral myocarditis, and 36 normal healthy children were enrolled as control. CK-MB was also measured in all subjects enrolled into the study. Results The serum levels of HMGB1, VE-cadherin and CK-MB in children with acute stage viral myocarditis (HMGB1 :[5.14 ±0. 23] mg/L;VE-cadherin: [5.36 ±0. 92] mg/L;CK-MB: [31.42 ± 3.22] U/L)were significantly higher than those with recovery stage viral myocarditis ( HMGB1: [ 0. 92 ± 0. 14 ] mg/L, VE-cadherin: [2. 93 ±0. 64] mg/L; CK-MB: [ 13.75 ± 3.18 ] U/L) ( t = 11.37,10. 26 and 12. 17 respectively ,Ps ＜ 0. 01 )and control (HMGB1 :[ 0. 86 ± 0. 12 ] mg/L; VE-cadherin: [ 2. 86 ± 0. 65 ] mg/L; CK-MB: [ 12. 83 ±3.04] U/L) (t = 12.06,11.19 and 12. 64 respectively,Ps ＜0.01 ). However,we found no significant difference in the serum levels of HMGB1, VE-cadherin and CK-MB between recovery stage viral myocarditis group and the control ( t = 1.26,1.19,1.43, Ps ＞ 0. 05 ). There were positive correlations between HMGB1 and VE-cadherin,CK-MB (r = 0. 73,0. 79, Ps ＜ 0. 05 ) ;and positive correlation between VE-cadherin and CK-MB (r= 0. 82, P ＜0.05). Conclusion HMGB1 and VE-cadherin may play roles in the viral myocarditis pathogenesis, which can be new prognosis factors for viral myocarditis.
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Objective To observe the changes of serum vascular endothelium-cadherin (VE-cadherin) and plasma arginine vasopressin (AVP) in patients with acute brain injury, and to investigate the VE-cadherin and plasma AVP related to brain edema and prognosis following acute brain injury. Method ELISA was used to measure the serum VE-cadherin level and radioimmunoassay was used to measure serial concentration of AVP in plasma of 110 patients with acute brain injury 12 hours and 3 and 5 days after brain injury. The outcome was estimated in combination of clinical manifestation, iconographic findings and Glasgow Outcome Scale (GOS). Results The levels of VE-cadherin and AVP increased significantly over 12 hours after brain injury with reduction of GCS. The severer brain injury resulted in higher levels of VE-cadherin and AVP, and prolongation of peak brain edema (P < 0.01). VE-cadherin levels showed positive correlation with severity of brain edema (r = 0.69, P < 0.01) and prolongation of brain edema (r = 0.70.P < 0.01). AVP levels had positive correlation with severity of brain edema (r = 0.65, P < 0.01) and prolongation of brain edema (r = 0.64, P < 0.01). Furthermore, there were significant increases in VE-cadherin and AVP levels in patients with low GOS groups (P < 0.01). The VE-cadherin and AVP leveb in poor outcome group persistently increased for 5 days after brain injury and were higher than those in good outcome group and normal control group. There were a correlation between VE-cadherin and AVP in patients with acute brain injury (r = 0.75, P < 0.01). Conclusions VE-cadherin and AVP leveb are closely associated with the prognosis of patients after acute brain injury. VE-cadherin and AVP may play pivotal roles in the pathogenesis of brain edema after acute brain injury.
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Objective To investigate the changes and clinical implications of VE-eadherin during the courseof progressive cerebral infarction.Method One hundred sixty-seven patients with acute cerebral infarction of Rong Jun General Hospital of Shandong Province and the Central People' s Hospital of Tengzhou were diagnosed in our hospital from May 2006 to July 2007,were diagnosed according to the ill%gnome criteria set by the 4th national cerebrovascular disease conference in 1995.Of them there were 102 ases with progressive cerebral infarction patients and 65 cases with non-progressive cerebral infaction.The progressive cerebral infarction patients were divided into 3 groups according to Pullicino's expressions:the big infarction focus(32 patients) ,the medium-sized infarction focus(34 patients) and the small infarction focus(36 patients) .The neurological deficits were divided into 3 groups according to the crrteria set by the 4th national cerebrovascular disease conference in 1995:light-defictits(38 patients),the moderate dificits (32 patients) and sever ditlcits (32 patients).The 65 non-progressive irffarction patients were stable without headache,vertigo and tinnitus.Arother 60 healthy subjects were entered as control group.Blood samples of all the patients' were collected at 0 h,24 h,3 d,7 d,14 d,24 d and the serum VE-eadherin by ELISA method was asaayed.All the data were analyzed by SPSS 10.0 software and One-way ANOVA was applied to intergroup comparisons for mote than two groups.Results The VF-cadherin level of patieras with progressive infarction increased in acute stage,reached the peak 3 days after onset,declined remarkably 7 days later and got nearly normalized within 21 days.The results were significantly different from those of non-progressive and controlgroup(P＜0.01).The VE-cadherin concentration was higher in patients with bigger size infarction and more sever symptoms.Conclusions The VE-cadherin level is related to the infarction size,course and the severity,and higher in the progressive group.VE-cadherin could be used for predicting prognostic of cerebral infarction and clinically valuable for treating ischemie cerebrovascular disease.