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Objective:To study the effects of endothelial progenitor cells (EPC)-derived exosomes on hyperoxia-induced injury in type Ⅱ alveolar epithelial cell (AECⅡ) in neonatal rats.Methods:EPCs of rats were cultured and exosomes were collected using Total Exosome Isolation kit. Primary cultured AECⅡof neonatal rats were randomly assigned into three groups: the control group, the hyperoxia group and the exosome group. The control group was cultured in room air with 5%CO 2, the hyperoxia group was cultured in 95%O 2 with 5%CO 2 and the exosome group was cultured with 0.1 mg/ml EPC-derived exosomes in 95%O 2 with 5%CO 2. Cell viability was detected using cell counting kit-8 (CCK-8) and apoptosis was detected using flow cytometry on d2, d4, and d6. Results:EPC-derived exosomes isolated from EPC culture supernatant were confirmed morphologically using transmission electron microscopy. After co-incubation of Dil-labeled EPC-derived exosomes with AEC Ⅱ for 24 h, Dil fluorescence was detected in the cytoplasm of AEC Ⅱ, indicating exosomes were uptaken by AEC Ⅱ. Compared with the control group, hyperoxia decreased cell viability and increased apoptosis of AEC Ⅱ and the injury was aggravated with the prolongation of hyperoxia duration ( P<0.001). Cell injury in the exosome group was milder than the hyperoxia group ( P<0.001). Compared with the control group, cell viability on d4 and d6 of hyperoxia was lower ( P=0.029 and 0.005 respectively) and cell apoptosis at d6 of hyperoxia was higher in the exosome group ( P=0.007). Conclusions:EPC-derived exosomes may partially attenuate hyperoxia-induced cell injury in neonatal rat AEC Ⅱ.
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Objective:To investigate the effect of Astragaloside Ⅳ-mediated Endothelial progenitor cells derived exosomes (EPC-Exos) on the biological function of EPC-Exos damaged by high glicose.Methods:EPCs from human umbilical cord blood were isolated and cultured in vitro. the EPC-Exos secreted by EPCs were extracted by ultracentrifugation combined with ultrafiltration, and identified by specific markers CD9, CD63 and CD81, respectively. After the cells were cultured for 24 hours with AS-IV at 100 mg/L and PBS at the same volume, the morphological characteristics of EPC-Exos were observed by transmission electron microscope. Human endothelial cells were isolated, cultured and identified in vitro. The identified endothelial cells were pretreated with 30 mmol/L glucose for 120 h and randomly divided into experimental group and control group, at the same time set the normal group. The cells were cultured for 24 hours, the effects of EPC-Exos on proliferation, adhesion, migration and angiogenesis of endothelial cells damaged by high glucose were observed by using cell counting kit-8 (CCK-8) Cell Proliferation Assay Kit, cell scratch test, adhesion assay and in vitro angiogenesis assay by Matrigel. Results:Compared with the normal group, the proliferation, migration, adhesion and tubulogenesis of human endothelial cells in the control group were significantly lower ( t=24.35, 6.80, 10.65, 9.62, P<0.05). Compared with the control group, the proliferation, adhesion, migration and tubulogenesis of human endothelial cells in the experimental group were significantly enhanced ( t=30.68, 5.99, 5.40, 8.25, P<0.05). Conclusions:EPC-Exos mediated by AS-Ⅳ can significantly improve the biological function of human endothelial cells damaged by high glucose and has the potential to modulate endothelial neovascularization in diabetic rats.
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Objective:To investigate the effect of Danggui Buxuetang（DGBX）on the functional activity of rat endothelial progenitor cells（EPCs）exposed to different luminar shear stress （SS）. Method:EPCs isolated from rat bone marrow were incubated on a parallel plate flow chamber at a steady SS of 0， 0.12， 1.2， 2.4 Pa for 6 h，then the cells exposed to different SS were randomly divided into 8 groups： control group （perfused with serum free medium），simvastatin group（0.1 μmol·L<sup>-1 </sup>simvastatin），3 DGBX groups（low，medium，high-dose DGBX）and 3 inhibitor groups（3 DGBX groups with LY294002）. After 12 h，the samples were collected for the detection of cell proliferation ，migration，tubule formation ，the secretion of nitric oxide （NO） ，and the expressions of endothelial nitric oxide synthase（eNOS） mRNA and protein kinase B（Akt），respectively. Result:Compared with the control group，simvastatin and DGBX（high-dose）could both promote the functional activities and NO secretion，and up-regulate the expressions of eNOS mRNA and Akt protein in EPCs exposed to different SS（<italic>P</italic><0.05），while DGBX（mid-dose）could do these only at 0 Pa. However，LY294002 could inhibit all effects of DGBX on EPCs. Conclusion:SS seems to play an important role in the effect of DGBX on EPCs，and DGBX could promote the functional activity of EPCs exposed to SS by up-regulating the expressions of NO/eNOS/Akt.
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OBJECTIVE: To discuss the effect of feiliuping ointment on proliferation, angiogenesis and PI3K/AKT pathway of rat bone marrow-derived endothelial progenitor cells (EPCs) in A549 lung tumor cells supernatant. METHODS: Bone marrow mononuclear cells were isolated from Sprague-Dawley rats by density gradient centrifugation methods and cultured in EGM-2 MV medium. And fluorescent immunocytochemistry was used to detect CD133 and vascular endothelial growth factor receptor 2 (VEGFR2) expression. EPCs were treated with 0.9% sodium chloride solution (blank control) or serum of rats treated with high dose and low dose Feiliuping ointment for 48 h to explore the concentration-effect, or with serum of rats treated with high dose feiliuping ointment for 0, 12, 24, 48 h to explore the time-effect. MTT assay and Matrigel method were used to detect the proliferation and angiogenesis of EPCs. The NF-κB, p-Akt, PI3K p85 proteins of EPCs were detected by Western blot. RESULTS: After induced culture for 7 d, the isolated bone marrow mononuclear cells exhibited round or short spindle-shaped appearance, were Dil-Ac-LDL and FITC-UEA-1 positive, and expressed CD133+VEGFR2+. After being treated with high dose feiliuping ointment serum for 48 h, OD490 nm value was lower than that in the control group or low dose feiliuping ointment group (P<0.05). The inhibition effect of rat serum after Feiliuping ointment administration on EPCs was dose-dependent and time-dependent. Matrigel tube formation assays showed that the counts of tube formation of EPCs in high dose Feiliuping ointment group was lower than that in control group or low dose of feiliuping ointment group (P<0.05). And the levels of NF-κB, p-Akt, PI3K p85 proteins in high dose of feiliuping ointment group were lower than those in control group(P<0.01). CONCLUSION:S Treatment with administration can Feiliuping ointment serum would inhibit the proliferation and tube formation of bone marrow-derived EPCs in A549 supernatant via the down-regulation of PI3K/AKT signaling pathway.
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Objective: To investigate the effect of poly (L-lactic acid caprolactone) (PLCL) /gelatin electrospinning on the angiogenesis differentiation of endothelial progenitor cells (EPCs). Methods: Rat bone marrow-derived EPCs were isolated and cultured, then identification was performed. After preparation of PLCL/gelatin blend electrospun scaffold, scanning electron microscopy and water contact angle test were carried out. EPCs were grown on PLCL/gelatin electrospinning and CCK8 was used to detect cell proliferation. The expression of vascular endothelial growth factor (Vegf ) and kinases insert region receptor (Kdr) was observed by RT-PCR and the expression of VEGF protein was observed by Western blotting. Results: The density gradient centrifugation combined with differential adherence method could effectively isolate EPCs. PLCL/gelatin electrospun nanofibers were porous, and the hydrophilic properties were favorable for cell adhesion, and EPCs grew well on the scaffold. The expression of Vegf and Kdr gene in PLCL/gelatin group was higher than that in control group (P=0.000), and the expression of VEGF protein was also increased (P=0.000). Conclusion: PLCL/gelatin is an ideal scaffold for tissue engineering, and it can promote the angiogenesis differentiation of EPCs.
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Objective · To investigate the effect of poly (L-lactic acid caprolactone) (PLCL)/gelatin electrospinning on the angiogenesis differentiation of endothelial progenitor cells (EPCs).Methods· Rat bone marrow-derived EPCs were isolated and cultured,then identification was performed.After preparation of PLCL/gelatin blend electrospun scaffold,scanning electron microscopy and water contact angle test were carried out.EPCs were grown on PLCL/gelatin electrospinning and CCK8 was used to detect cell proliferation.The expression of vascular endothelial growth factor (Vegf) and kinases insert region receptor (Kdr) was observed by RT-PCR and the expression of VEGF protein was observed by Western blotting.Results· The density gradient centrifugation combined with differential adherence method could effectively isolate EPCs.PLCL/gelatin electrospun nanofibers were porous,and the hydrophilic properties were favorable for cell adhesion,and EPCs grew well on the scaffold.The expression of Vegfand Kdr gene in PLCL/gelatin group was higher than that in control group (P=0.000),and the expression of VEGF protein was also increased (P=0.000).Conclusion · PLCL/gelatin is an ideal scaffold for tissue engineering,and it can promote the angiogenesis differentiation of EPCs.
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Introduction@#Endothelial progenitor cells (EPC) have a role in the maintenance and promotion of vascular repair and are negatively correlated with coronary atherosclerosis. @*Goal@#To culture of EPC-CFUs during coronary atherosclerosis, evaluate endothelial nitric oxide synthetase (eNOS) enzyme levels in the culture.@*Materials and Methods@#The 10 ml blood was drawn from the peripheral vein of 12 man patients that stable angina 4, acute myocardial infarction (AMI) 4 and healthy people 4. Peripheral blood mononuclear cells were isolated by Ficoll density-gradient centrifugation and EPC-CFUs was assayed after two platings and a 6 day culture on fibronectin coated, 72 well plates, as described. eNOS enzyme titers were determined by ELISA according to the protocol in the cells culture.@*Results@#The people were 52±2.12 years. The number of EPC-CFUs increases with accordance of patients with stable angina, AMI, healthy people with the statistical significance (F=17.3, p<0.001): stable angina (2.6±0.47 colony/well), AMI (6.7±0.81 colony/well), healthy people (10.5±1.34 colony/well). Furthermore, ANOVA test of eNOS enzyme levels in patients with stable angina (5.2±0.61 pg/ml), AMI (8.7±1.49 pg/ml) and healthy people (13.7±2.48 pg/ml). The significant difference (F=6.2, p=0.003) was observed among the three groups. The number of EPC-CFUs had direct significantly correlation (r=0.621, p<0.001) with the eNOS enzyme levels of this culture.@*Conclusion@#Number of EPC-CFUs and eNOS enzyme levels decrease at patient with stable angina, indicate more than endothelial dysfunction.@*Ethical approval@#The ethics committee of Mongolian National University of Medical Sciences (ID: 6/3/201506, approved on Jan 01, 2015)
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Objective To investigate the protective effects of vascular endothelial growth factor-165 (VEGF165) transfected the endothelial progenitor cells (EPCs) mediated by lentivirus on acute lung injury (ALI) in rats. Methods The mononuclear cells from the male Sprague-Dawley (SD) rats were isolated and cultured to get the EPCs for study. The lentivirus vector carrying the human VEGF165 gene was constructed. According to the random number table method, 90 male SD rats were divided into ALI model group, phosphate buffer solution (PBS) group, EPCs treatment group, none transfected EPCs treatment group and VEGF165 transfected EPCs treatment group, and the rats in each group were subdivided into 4, 12 and 48 hours subgroups, with 6 rats in each subgroup. The rat model of ALI was reproduced by intravenous injection of oleic acid (0.15 μL/g). Then each treatment group was given PBS, EPCs, none transfected EPCs and VEGF165 transfected EPCs respectively with the same volume of 0.2 mL. For the groups with cells, about 1×106cells were contained. Abdominal aortic blood and lung tissue were harvested at 4, 12 and 48 hours. Arterial blood gas analysis was performed. The lung wet/dry weight ratio (W/D) was calculated. The expressions of induced nitric oxide synthase (iNOS), endothelin-1 (ET-1) and VEGF165 were determined by enzyme-linked immunosorbent assay (ELISA). After dyed with hematoxylin-eosin (HE), the lung tissue pathology was observed and the lung injury score was performed. Results Compared with the ALI model group, the arterial partial pressure of oxygen (PaO2) in EPCs, none transfected EPCs and VEGF165 transfected EPCs treatment groups was significantly increased from 4 hours, and lung W/D, expressions of iNOS and ET-1 were significantly decreased, and VEGF165 expression was significantly increased. Compared with the EPCs treatment group, the increase in PaO2, the decrease in lung W/D and expressions of iNOS and ET-1, and the increase in VEGF165 expression in VEGF165 transfected EPCs treatment group were more significant [4 hours: PaO2(mmHg, 1 mmHg = 0.133 kPa) was 82.84±10.69 vs. 72.34±9.36, lung W/D ratio was 4.83±0.23 vs. 5.55±0.37, iNOS (ng/mg) was 8.77±1.10 vs. 14.84±1.34, ET-1 (ng/mg) was 103.41±5.66 vs. 153.08±5.12, VEGF165 (ng/mg) was 130.56±12.16 vs. 83.03±5.95; 12 hours: PaO2(mmHg) was 91.67±6.81 vs. 78.5±8.81, lung W/D ratio was 4.44±0.35 vs. 5.32±0.25, iNOS (ng/mg) was 7.23±0.24 vs. 14.04±1.18, ET-1 (ng/mg) was 91.98±3.52 vs. 125.99±7.55, VEGF165 (ng/mg) was 164.49±5.71 vs. 96.61±6.12]; individual parameters reached valley value or peak value at 48 hours [lung W/D ratio was 4.26±0.30 vs. 4.89±0.15, iNOS (ng/mg) was 5.79±0.85 vs. 12.72±1.10, ET-1 (ng/mg) was 74.53±7.10 vs. 108.33±5.84, VEGF165 (ng/mg) was 237.43±10.79 vs. 134.24±11.99, all P < 0.05]. Over time, lung tissue injury in each group was gradually increased, and the lung injury score was gradually increased. The lung injury score at 48 hours in the EPCs, none transfected EPCs and VEGF165 transfected EPCs treatment groups were lower than that in the ALI model group. Compared with the EPCs treatment group, the VEGF165 transfected EPCs treatment group had a lower score at 48 hours (8.50±1.05 vs. 10.50±1.05, P < 0.05). Conclusion The transplantation of EPCs which were transfected with VEGF165 mediated by lentivirus could obviously improve the oxygen pressure, reduce the lung water seepage, decrease the iNOS and ET-1 expressions in lung tissue, and had obvious protective effects on ALI.
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Objective·To study the change of circulating endothelial progenitor cells (cEPCs) in acute ischemic stroke (AIS) patients within one week after attack,and the correlation of cEPCs with the prognosis.Methods·Ninty-two patients with AIS (AIS group) and 20 patients with risk factors (Risk group) were recruited.The proportion of cEPCs (CD34TKDR+ cells) in peripheral blood mononuclear cells of AIS patients was measured by flow cytometry (FCM) on the first day of admission and the seventh day after attack.Functional recovery was assessed by modified Rankin Scale (mRS) on the 90th day after onset.The cEPCs percentages of AIS patients with different mRS were compared to analyze their correlation.Results·Compared with Risk group,cEPCs percentage of AIS group on the 1st day of admission was lower (P=0.016).In AIS group,compared with poor prognosis group (mRS＞2),eEPCs percentage of good prognosis group on the 7th day after onset (mRS ≤ 2) elevates (P=0.002).The result of multiple linear regression showed that cEPCs percentage on the 7th day after onset was positively correlated with mRS on the 90th day (t=4.608,P=0.011).Conclusion·The percentage of cEPCs in peripheral blood of AIS patients decreases significantly during the acute phase.The percentage on the 7th day after onset is correlated with prognosis of AIS patients.
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Objective The extracorporeal cardiac shock wave therapy ( CSWT ) can promote angiogenesis in ischemic myo-cardium and improve myocardial perfusion , but its mechanisms remain to be clarified .This study aimed to explore the effects of CSWT on the endothelial progenitor cells (EPCs), vascular endothe-lial growth factor ( VEGF) and Interleukin-8 ( IL-8) as well as its re-lieving effect on angina pectoris in patients with coronary heart dis-ease. Methods After Dobutamine stress echocardiography ( DSE) and 99 mTc-MIBI myocardial perfusion imaging (MPI) at rest and un-der stress, 25 patients with coronary heart disease underwent 9 three-month cycles of CSWT .Before and after the treatment , we obtained the results of 6-min walk test, NYHA cardiac function grades , CCS angina pectoris classes, Seattle Angina Questionnaire (SAQ) scores, doses of nitroglycerin administered , left ventricular diastolic di-ameter ( LVDD) , and left ventricular ejection fraction ( LVEF) .We evaluated myocardial perfusion and myocardial contractile function using MPI and the peak systolic strain rate (PSSR) at rest and under stress, respectively. Results After CSWT, the numbers of EPCs and EPC-CFUs cultured in vitro were significantly increased as compared with the baseline (34.52±6.58 vs 19.56±4.28, P0.05).The PSSR showed no significant changes at rest (1.21±0.62 vs 1.04±0.43, P>0.05) but remark-ably increased under stress after CSWT (2.02±1.00 vs 1.35±0.66, P<0.01). Conclusion CSWT can up-regulate the expressions of VEGF and IL-8 and improve the function of EPCs in the peripheral blood , and thus plays an important role in relieving the symptoms of angina pectoris , promoting cardiac function and enhancing exercise tolerance in patients with coronary heart disease .
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Purpose To clarify the role of KAI1/CD82 in metastasis of nasopharyngeal carcinom and to evaluate the clinical efficacy of KAI1/CD82-expressing EPCs in the prevention of nasopharyngeal carcinoma.Method Umbilical vein-derived EPCs were infected with KAI1/CD82-expressing lenti-virus to get a KAI1/CD82-overexpressing EPC cell line (KAI1/CD82-EPCs).A xenograft mouse model of human nasopharyngeal carcinoma was established,and KAI1/CD82-EPCs were injected through the tail vein.The effect of the KAI1/CD82-EPCs on growth and metastasis of the xenograft was observed.Results Time required for tumor formation was 14.70 ± 3.81,15.05 ±3.85,14.20 ± 3.55 days respectively for the EPCs,EPCs-NC,and KAI1/CD82-EPCs groups,with no significant difference among the three groups (P =0.771).Weight of the xenograft was (1.388 ±0.204) g,(1.487 ±0.223) g,(1.485 ±0.234) g respectively for the EPCs,EPCs-NC,and KAI1/CD82-EPCs groups,with no significant difference (P =0.274).Rate of lung metastasis was 55％,45％ and 10％ for the EPCs,EPCs-NC,and KAI1/CD82-EPC groups,and the difference was significant (P =0.005).Number of metastatic lesions was 34.27 ± 5.35,38.44 ± 9.63,17.50 ± 3.54 for the three groups,and the difference was also significant (P =0.007).Immunohistochemistry indicated positive KAI1/CD82 expression in metastatic lesion of the KAI1/CD82-EPCs group,but no KAI1/CD82 expression in the EPCs group or EPCs-NC group.Conclusion KAI1/CD82-expressing EPCs inhibits lung metastasis of the xenograft mouse model of human nasopharyngeal carcinoma.
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Dysfunction or loss of blood vessel causes several ischemic diseases. Although endothelial progenitor cells (EPCs) are a promising source for cell-based therapy, ischemia-induced pathophysiological condition limits the recovery rate by causing drastic cell death. To overcome this issue, we attempted to develop a cell-targeted peptide delivery and priming system to enhance EPCbased neovascularization using an engineered M13 bacteriophage harboring nanofibrous tubes displaying ∼ 2700 multiple functional motifs. The M13 nanofiber was modified by displaying RGD, which is an integrin-docking peptide, on the minor coat protein, and bymutilayering SDKPmotifs,which are the key active sites for thymosin b4, on themajor coat protein. The engineered M13 nanofiber dramatically enhanced ischemic neovascularization by activating intracellular and extracellular processes such as proliferation, migration, and tube formation in the EPCs. Furthermore, transplantation of the primed EPCs with the M13 nanofiber harboring RGD and SDKP facilitated functional recovery and neovascularization in a murine hindlimb ischemia model. Overall, this study demonstrates the effectiveness of theM13 nanofiber-based novel peptide deliveryandprimingstrategy inpromotingEPC bioactivity and neovessel regeneration. To our knowledge, this is first report onM13 nanofibers harboring dual functional motifs, the use of which might be a novel strategy for stem and progenitor cell therapy against cardiovascular ischemic diseases.
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Animals , Bacteriophages , Blood Vessels , Catalytic Domain , Cell Death , Endothelial Progenitor Cells , Hindlimb , Ischemia , Nanofibers , Regeneration , Stem Cells , ThymosinABSTRACT
Endothelial dysfunction in atherosclerosis is associated with increasing oxidative stress that could be reversed by antioxidant. Therefore epigallocatechin gallate (EGCG), epicatechin gallate (ECG), epigallocatechin (EGC) and catechin (C) of tea flavonoids were investigated for their roles in regenerating endothelial cell. Peripheral blood mononuclear cells (PB-MNCs) were isolated, plated and cultured in medium with/without treatment of EGCG, ECG, EGC and C. Results showed that among all EGCG, ECG, EGC and C concentrations tested, 12.5 µmol/L was not cytotoxic for peripheral blood-derived endothelial progenitor cells (PB-EPCs). Treatment of EGCG, ECG, EGC or C increased the percentages of CD34, CD133, VEGFR-2 expressions and suppressed hydrogen peroxide-induced percentages of reactive oxygen species (ROS) level in PB-EPCs. Taken together, our current results showed that EGCG, ECG, EGC or C of tea flavonoids could induce differentiation of PB-MNCs into PB-EPCs as well as protect PB-EPCs from oxidative damage by suppresing the intracellular ROS levels.
Subject(s)
Apoptosis , Atherosclerosis , Catechin , Electrocardiography , Endothelial Cells , Endothelial Progenitor Cells , Flavonoids , Hydrogen , Oxidative Stress , Reactive Oxygen Species , Tea , Vascular Endothelial Growth Factor Receptor-2ABSTRACT
Objective To measure the level of circulating endothelial progenitor cell (EPC) and serum vascular endothelial growth factor (VEGF), and to investigate its clinical significance in patients with different stages of renal cell carcinoma (RCC). Methods The level of circulating EPC was quantified by assaying CD45-CD34+VEGFR-R2 +cell phenotype in 45 patients with RCC (RCC group), 30 patients with benign renal tumors(benign renal tumors group) and 30 healthy controls (control group). Serum VEGF was quantified by enzyme- linked immunosorbent assay (ELISA). The results were compared. Results The level of EPC in RCC groups was (0.265 ± 0.042)%, in benign renal tumors group was(0.053 ± 0.008)% , and in control group was (0.048 ± 0.006)%. The level of EPC in RCC group was significantly higher than that in benign renal tumors group and control group (P<0.05). The level of EPC in Ⅲ- Ⅳ stage patients was significantly higher than that in Ⅰ-Ⅱ stage patients:(0.312 ± 0.038)%vs. (0.215 ± 0.021)%, P<0.05. Three months after operation, the level of EPC in 33 RCC patients without pretreatment was (0.078 ± 0.003)% and significantly lower that before treatment (P<0.05). The level of VEGF in RCC groups was (305.5 ± 29.1) ng/L, in benign renal tumors group was (29.8 ± 3.2) ng/L, and in control group was (25.1 ± 2.8) ng/L. The level of VEGF in RCC group was significantly higher than that in benign renal tumors group and control group (P<0.05). The level of VEGF inⅢ-Ⅳstage patients was significantly higher than that inⅠ-Ⅱstage patients:(365.6 ± 34.6) ng/L vs. (256.2 ± 23.2) ng/L, P<0.05. Pearson association analysis showed that the level of EPC had positive associations with VEGF (r=0.714, P<0.01). Multiple linear regression showed that the size of kidney neoplasms was a dependent factor for the level of EPC. Conclusions The level of EPC has a positive association with VEGF. EPC maybe a new biomarker for RCC.
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Objective To explore the angiogenic function of EPC from peripheral blood in kidney transplanted patient and to reveal its regulative mechanism.Methods 23 chronic renal failure patients without diabetes were recruited in department of Urology Peking University Third Hospital from January 2014 to February 2015.Fasting peripheral blood mixed with heparin (20 U/mL) was collected one day before and 24 hours after kidney transplantation.We set preoperative blood as control and the postoperative blood as the experimental group.EPC from peripheral blood were isolated by density-gradient centrifugation.FACS was used to identify the EPC.The AA metabolites PGE2 in EPC cultured medium was measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS).Q-PCR and WB were used to detect the expression of endothelial markers in HUVEC cultured under the EPC conditional medium.Tube formation assay was performed to assess the angiogenic ability of HUVEC.Results EPC from kidney transplantation expressed c-kit and CD31 by FACS analysis.Multiple types of AA metabolites was detected in the conditional medium by LC-MS/MS and level of PGE2 increased into two folds after kidney transplantation, compared with that before operation(P ＜ 0.05).HUVEC highly expressed CD31 and VE-cadherin cultured under conditional medium, which were 1.5 folds compared with that before operation (P ＜ 0.01).And those cells formed more tubes than that in control group, which showed better angiogenic capacity.HUVEC, treated by PGE2, had the similar biological characteristics like the conditional culture.Conclusions EPCs in the peripheral blood form kidney transplantation patient secret the PGE2, which can enhance the capacity of angiogenesis in HUVEC.
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@#Endothelial nitric oxide synthase (eNOS) and nitric oxide (NO) can adjust the recovery of blood flow after cerebral ischemia, inhibit inflammatory reaction and promote the regeneration of the nerve and blood vessels. The gene polymorphism of eNOS is closely related to the onset of ischemic cerebrovascular disease. This article reviewed the relationship between eNOS and ischemic cerebrovascular disease.
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Objective To study the modulation in number and function of endothelial progenitor cell ( EPC ) in multiple organ dysfunction syndrome ( MODS ) after trauma in swine, and to investigate its pathogenesis. Methods Forty pigs were divided into sham group and MODS group ( each, n = 20 ). The model of MODS of two-hit injury, namely hemorrhagic shock and endotoxemia, was reproduced. The peripheral blood was collected before hemorrhage ( T1 ) and endotoxin injection ( T2 ), and 1 hour ( T3 ), 24 hours ( T4 ), 48 hours ( T5 ) after endotoxin injection. Phosphorylation of p38 mitogen-activated protein kinase ( p-p38MAPK ) in mononuclear cell was determined by Western Blot, the content of tumor necrosis factor-α ( TNF-α) was determined with enzyme linked immunosorbent assay ( ELISA ), and the number of EPC was determined with flow cytometry. Results Model of MODS was successfully reproduced in 17 pigs. In model group, the expression of p-p38MAPK ( A value ) peaked at T3 ( 4.83±0.52 ), and gradually declined at T4 and T5 ( 4.36±0.43, 1.93±0.33 ), and the expression of p-p38MAPK at T3-T5 was significantly higher than that at T1 ( 1.00±0.22, all P<0.01 ). The plasma concentration of TNF-α( ng/L ) at T3 in MODS group was obviously elevated compared with that of sham group ( 532.43±52.17 vs. 129.03±20.45, t=31.163, P<0.001 ), and it peaked at T3, it then gradually lowered, and it was significantly higher at T4 and T5 than that in sham group ( T4: 398.93±35.75 vs. 131.12±29.53, t = 26.562, P < 0.001; T5: 287.48±27.26 vs. 126.44±26.96, t=17.861, P<0.001 ). The number of EPC ( ×107/L ) was apparently increased in MODS group at T3 compared with sham group ( 4.832±0.624 vs. 3.545±0.363, t=9.542, P<0.001 ), and it peaked at T3, then gradually decreased, and the number of EPC at T4 and T5 was significantly lower than that in sham group ( T4:2.628±0.627 vs. 3.442±0.325, t=5.043, P<0.001;T5:2.203±0.711 vs. 3.471±0.323, t=2.972, P<0.001 ). Conclusion Phosphorylation of p38MAPK could increase the plasma concentration of TNF-αand decrease the quantity of EPC in MODS,which may be one of the mechanisms of MODS.
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Objective To investigate the role of endothelial progenitor cells ( EPCs ) transplantation in rats with sepsis induced by endotoxin ( lipopolysaccharides, LPS ). Methods Sixty clean grade Sprague-Dawley ( SD ) rats with genetic background were divided into three groups according to random number table method:control group, model group, and EPCs transplantation group, with 20 rats in each group. The sepsis model was reproduced by intravenous delivery of LPS 5 mg/kg. Rats in control group were injected with the same amount of normal saline. EPCs were isolated, and cultured and identified were fluorescently labeled with the green fluorescent protein ( GFP ) adenoviral transfection method. The EPC transplantation group was injected with LPS, then a fluorescently labeled EPCs suspension was injected via the tail vein 1 hour later. The expression of fluorescent markers of EPCs was detected with both small animal in vivo imaging instrument and frozen section. Seven days after transplantation, abdominal aorta blood was collected to determine interleukins ( IL-6 and IL-10 ) in peripheral blood with enzyme linked immunosorbent assay ( ELISA ), and the lung, liver, and kidney tissues were harvested, the wet/dry ratio of the lung ( W/D ) was calculated, and hematoxylin and eosin ( HE ) staining was performed to observe, the change in histopathology. Toll-like receptor 4 ( TLR4 ) mRNA expression in lung, liver, and kidney tissues was determined with real-time reverse transcription-polymerase chain reaction ( RT-PCR ). Results The positive rate of EPCs cells with double marking of CD133 and CD34 was 99.0% at the 5th generation of subculture by using flow cytometry. After the transplantation of EPCs labeled with the green fluorescent protein, the appearance of fluorescence indicated that EPCs were mainly localized in the chest, and a stronger fluorescence was observed near the blood vessels. EPCs transplantation could significantly reduce the inflammatory cell infiltration and cell damage in lung, liver, and kidney tissue in septic rats. Compared with control group, the expression of IL-6 and IL-10 in the peripheral blood, W/D ratio, and TLR4 mRNA in lung, liver, and kidney were increased significantly in the model group. Compared with model group, the expressions of IL-6 and IL-10 in the peripheral blood were significantly reduced after EPCs transplantation [ IL-6 (μg/L ):2.127±0.118 vs. 2.664±0.438, IL-10 ( ng/L ): 24.5±3.9 vs. 31.5±3.8, both P < 0.01 ]. EPCs transplantation reduced the W/D ratio of lung, liver and kidney tissues ( lung: 4.68±0.24 vs. 5.48±0.15, liver: 3.33±0.11 vs. 3.94±0.09, kidney: 4.08±0.20 vs. 4.84±0.21, all P < 0.05 ], and down-regulated the expression of TLR4 mRNA ( ×103, lung: 782±131 vs. 1 136±126, liver: 39.1±14.0 vs. 69.2±8.7, kidney: 52.2±15.2 vs. 83.5±17.1, all P < 0.01 ). Conclusions EPCs can enter the lung, liver and kidney tissues of the rat successfully after transplantation of EPCs via vein. EPCs transplantation can down-regulate pro-inflammatory process, help to recover the balance of pro-and anti-inflammatory processes, alleviate the damage to the lung, liver, and kidney tissue significantly.
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Various approaches have been attempted in translational moyamoya disease research. One promising material for modeling and treating this disease is vascular progenitor cells, which can be acquired and expanded from patient peripheral blood. These cells may provide a novel experimental model and enable us to obtain insights regarding moyamoya disease pathogenesis. We briefly present the recent accomplishments in regard to the studies of vascular progenitor cells in moyamoya disease.
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Humans , Models, Theoretical , Moyamoya Disease , Stem CellsABSTRACT
OBJECTIVE: To investigate the neuroprotective role of early statin treatment on stroke patients.