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Immature hematopoietic progenitors are a constant source for renewal of hemocyte populations and the basic component of the tissue and cell repair apparatus. A unique property of these cells of internalizing extracellular double-stranded DNA has been previously shown. The leukostimulatory effect demonstrated in our pioneering studies was considered to be due to the feature of this cell. In the present research, we have analyzed the effects of DNA genome reconstructor preparation (DNAgr), DNAmix, and human recombinant angiogenin on both hematopoietic stem cells and multipotent progenitors. Treatment with bone marrow cells of experimental mice with these preparations stimulates colony formation by hematopoietic stem cells and proliferation of multipotent descendants. The main lineage responsible for this is the granulocyte-macrophage hematopoietic lineage. Using fluorescent microscopy as well as FACS assay, co-localization of primitive c-Kit- and Sca-1-positive progenitors and the TAMRA-labeled double-stranded DNA has been shown. Human recombinant angiogenin was used as a reference agent. Cells with specific markers were quantified in intact bone marrow and colonies grown in the presence of inducers. Quantitative analysis revealed that a total of 14,000 fragment copies of 500 bp, which is 0.2% of the haploid genome, can be delivered into early progenitors. Extracellular double-stranded DNA fragments stimulated the colony formation in early hematopoietic progenitors from the bone marrow, which assumed their effect on cells in G0. The observed number of Sca1+/c-Kit+ cells in colonies testifies to the possibility of both symmetrical and asymmetrical division of the initial hematopoietic stem cell and its progeny.
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BACKGROUND: Diabetic wound healing disorder is one of the common complications of diabetes, but its pathogenesis has not yet been elucidated. Electrical stimulation therapy is one of the commonly used methods in the clinical treatment of wound injury, which can effectively promote the healing of injured skin. OBJECTIVE: To investigate the effects of electrical stimulation therapy on wound healing and angiogenesis in diabetic rats and its potential mechanism. METHODS: Forty-two SPF male Sprague-Dawley rats were enrolled in this study. A rat model of diabetic refractory wounds was established in 24 rats by tail vein injection of streptozotocin (50 mg/kg) combined with back skin wounds, and model rats were randomly divided into model group and electrical stimulation group. Meanwhile, normal rats were taken to cause back wounds as a blank control group. The electrical stimulation group was treated with electrical stimulation for 21 days. The blank control group and the model group were fed normally. The wound healing was evaluated on the 3rd, 7th, 14th and 21st days after treatment. After treatment, the rat serum and wound tissue were taken for index detection. The pathological morphology of rat wounds was observed by hematoxylin-eosin staining. The serum endothelial nitric oxide synthase, angiopoietin 1 and vascular endothelial growth factor levels were detected by enzyme-linked immunosorbent assay. The expression levels of angiopoietin receptor 2 and vascular endothelial growth factor receptor 2 proteins were detected by immunohistochemistry and western blot. An ethical approval was obtained from the Animal Experimental Ethics Committee of the Affiliated Hospital of Southwest Medical University. RESULTS AND CONCLUSION: The wound healing rate of diabetic rats was close to 90% on the 14th day after electrical stimulation, while the healing rate in the model group was less than 60% (P < 0.01). The serum endothelial nitric oxide synthase, angiopoietin 1 and vascular endothelial growth factor levels were significantly higher in the electrical stimulation group than the model group (P < 0.05 or P < 0.01). Meanwhile, compared with the model group, the electrical stimulation group had more neovascularization, larger vascular lumen, higher expression of angiopoietin receptor 2 and vascular endothelial growth factor receptor 2 around the blood vessels (P < 0.05 or P < 0.01). These findings indicate that electrical stimulation therapy can significantly promote wound healing and neovascularization in diabetic rats, and its mechanism is related to the increase of endothelial nitric oxide synthase, angiopoietin 1 and vascular endothelial growth factor levels and up-regulation of angiopoietin receptor 2 and vascular endothelial growth factor receptor 2 levels.
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Objective To explore whether liraglutide could affect high glucose induced-biological behavior of human endothelial colony forming cell (ECFC) by regulating sirtuin. Methods Mononuclear cells in human peripheral blood were isolated by density gradient centrifugation. ECFCs were induced and cultured with EBM-2 complete medium. The cells were randomly divided into three groups: normal glucose group (NG, 5.5 mmol/L D-glucose for 6 days), high glucose group (HG, 30 mmol/L D-glucose for 6 days), and osmotic pressure group (OSM, 5.5 mmol/L D-glucose+24.5 mmol/L mannitol for 6 days). The proliferation, migration, tube formation and senescence of ECFCs in the three groups were tested by EdU, Transwell, tube formation and SA-β-gal experiments respectively. The protein expression levels of Sirtuins (Sirtuin1-7), vascular endothelial growth factor (VEGF) and angiogenin were measured by Western blotting. Except NG and HG groups mentioned above, NG + liraglutide intervention group (5.5 mmol/L D-glucose+100 nmol/L liraglutide) and HG+liraglutide intervention group (30 mmol/L D-glucose+100 nmol/L liraglutide) were added in order to observe the effects of liraglutide on ECFC biological behavior and the expression of VEGF and angiogenin. The intervention time in all groups is 6 days. Results EdU, Transwell and tube formation experiments showed that the proliferation rate, migration ability and tube formation ability of ECFCs were lower in HG group [(30.16±12.36)%, 25.11±6.05, 27.50±3.90, respectively] than in NG group [(88.00±13.77)%, 64.89±10.73, 69.61±5.48, respectively], while the SA-β-gal experiment showed the senescence rate of ECFCs was higher in HG group than in NG group [(87.63±9.63)% vs. (71.35±7.58)%], all with statistical significance (P<0.05). Under high glucose conditions, the expression of Sirtuins family was generally reduced, and the expressions of VEGF and angiogenin significantly decreased (P<0.05). The expression of Sirtuin1 showed an upward trend with the increase of liraglutide concentration. After intervention with appropriate concentration of liraglutide, the expressions of VEGF and angiogenin significantly increased (P<0.05). EdU experiment showed the proliferation rate of ECFCs was higher in HG+liraglutide intervention group than in NG group [(54.09±27.29)% vs. (29.01±7.56)%], the Transwell and tube formation experiments showed that the migration ability and tube formation ability of ECFCs were higher in HG+liraglutide intervention group (32.25±4.99, 69.61±8.11) than in HG group (21.75±3.10, 39.85±5.78), and the SA-β-gal experiment showed the senescence rate of ECFCs was lower in HG+liraglutide intervention group than in HG group [(52.06±7.94)% vs. (69.03±1.57)%], all with statistical significance (P<0.05). Conclusions High glucose may accelerate ECFCs senescence while decrease the ability of proliferation, migration and tube formation of ECFCs, accompanied by down-regulation of Sirtuins, VEGF and angiogenin. Liraglutide may reverse the changes of ECFCs biological behavior induced by high glucose and up-regulate the expression of Sirtuin1, VEGF and angiogenin.
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Objective To investigate the effects of sevoflurane post treatment on cerebral ischemia-reperfusion injury in diabetic rats and non-diabetic rats.Methods Thirty-two male SD rats,aged 3.0-3.5 months,weighing 280-320 g,were divided into four groups by random number method (n =8):non diabetic ischemia reperfusion group (group NDC);diabetic ischemia reperfusion group (group DC2);non diabetic ischemia reperfusion group with the sevoflurane post-treatment (group NDS);diabetic ischemia reperfusion group with the sevoflurane post-treatment (group DS).The middle cerebral artery occlusion method and streptozotocin were used to establish the ischemiareperfusion injury model and diabetic model.2 h after ischemia and 24 h after reperfusion,the rats nerve defect scale was detected,the cerebral infarction volume was evaluated by TTC method,and Western blot method was used to detect angiogenin-1 (Ang 1) expression.Results Between the four groups of rats,nerve deficit score and infarct volume were significantly higher,Ang-1 protein relative expression was significantly lower in group DC than those in group NDC (P＜0.05);neural deficit score and infarct volume were significantly lower,Ang-1 protein relative expression was significantly higher in group NDS than those in group NDC (P＜0.05);nerve deficit score and infarct volume were significantly lower,Ang-1 protein relative expression was significantly higher in group DS than those in group DC (P＜0.05).Conclusion In the rats after cerebral ischemia reperfusion,diabetes could aggravate the nerve defect,increase the volume of cerebral infarction,reduce the expression of Ang-1;sevoflurane could reduce nerve defects,reduce infarct volume,increase the expression of Ang-1.The expression of Ang-1 and degree of injury after cerebral ischemia reperfusion in rats have relationship.
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Objective To investigate the effect of butylphthalide on the neurological function and the levels of (S-100B),homocysteine(Hcy),angiopoietin-1(ANG-1)in patients with acute cerebral infarction(ACI). Methods A total of 84 patients with ACI were selected from January 2016 to September 2017 in Henan Rongjun Hospital. The patients were divided into observation group and control group according to the treatment method,with 42 cases in each group. The patients in the two groups were treated with antithrombosis,blood sugar and blood pressure control,nutritional support,infection prevention and other conventional treatment measures. On the basis of routine treatment,the patients in the observation group were treated with butylphthalide injection 100 mL by intravenously guttae,twice a day for two weeks. The neurologic impairment of the patients in the two groups was assessed by the National Institutes of Health Stroke Scale(NIHSS)before and after treatment. The mental state and cognitive function of the patients in the two groups were scored by the mini-mental state examination(MMSE). The levels of serum S100B and ANG-1 were detected by Enzyme-linked immunosorbent assay. The level of Hcy was detected by AU480 automatic biochemical analyzer. The curative effect was evaluated after two weeks of treatment,and the adverse reac-tions of the patients were observed. Results There was no significant difference in the scores of NIHSS and MMSE between the two groups before treatment(P > 0. 05). The NIHSS score after treatment was significantly lower than that before treatment, and the MMSE score was significantly higher than that before treatment in the two groups(P < 0. 05). The NIHSS score in the observation group was significantly lower than that in the control group,and the MMSE score was significantly higher than that in the control group after treatment(P < 0. 05). There was no significant difference in serum S100B,Hcy and ANG-1 levels be-tween the two groups before treatment(P > 0. 05). The levels of serum S100B and Hcy after treatment were significantly lower than those before treatment,and the ANG-1 level was significantly higher than that before treatment in the tao groups(P <0. 05). The levels of serum S100B and Hcy in the observation group were significantly lower than those in the control group, and the ANG-1 level was significantly higher than that in the control group after treatment(P < 0. 05). The total effective rate in the observation group and the control group was 90. 48％(38 / 42)and 71. 43％(30 / 42)respectively,the total effective rate in the observation group was significantly higher than that in the control group(χ2 = 4. 659,P < 0. 05). The incidence of ad-verse reactions in the observation group and the control group was 11. 90％(5 / 42)and 9. 52％(4 / 42)respectively,there was no significant difference in the incidence of adverse reactions between the two groups(χ2 = 0. 092,P > 0. 05). Conclusion Butylphthalide can effectively regulate the levels of serum S100B,Hcy and ANG- 1,and improve the neurological function of patients with ACI.
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Objective:To research the clinical effect of HeDanPian combined with edaravone in the treatment of acute cerebral in farction and the influence on serum levels of interferon-γ (IFN-γ),promote angiogenin Ⅱ (Ang-2),homocysteine (Hcy).Methods:102 cases of patients with acute cerebral infarction were selected and randomly divided into the control group and the experimental group,with 51 cases in each group.The control group was treated with edaravone,intravenous infusion of30mg edaravone,1 time in the morning and night;The experimental group was treated based on the control group treated with HeDanPian,oral HeDanPian 14.6 g,1 time in the early to middle.The curative effect,nerve function defect score (NIHSS),serum IFN-γ Ang-2,Hcy,propylene glycol (MDA),superoxide disproportionation alcohol (SOD),interleukin 6,10 (IL-6,IL-10),tumor necrosis factor-α(TNF-α) levels,incidence of adverse reactions were compared between two groups.Results:After treatment,the total effective rate,serum Ang-2,SOD,IL-10 levels of experimental group were higher than those of the control group (P＜0.05),the NIHSS,serum IFN-γ,Hcy,MDA,IL-6,TNF-α levels of experimental group were lower than those of the control group (P＜0.05).No significant difference was found in the incidence of adverse reactions was found between the two groups (P＞0.05).Conclusion:HeDanPian combined with edaravone was effective in the treatment of acute cerebral infarction,which could regulate the expression of serum IFN-γ,Ang-2 and Hcy,improve the oxidative stress and inflammation.
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Objective To explore the relationship between angiogenin-1/2 (Ang-1/2) and clinical parameters of idiopathic pulmonary fibrosis (IPF), and to assess the value of Ang-1/2 in predicting the prognosis of patients with IPF.Methods A retrospective analysis was conducted. Ninety-one patients diagnosed as IPF by high resolution CT (HRCT) and lung biopsy admitted to Daqing Oil Field General Hospitalfrom March 2014 to January 2015 were enrolled. The general data, serum parameters and pulmonary function parameters of all patients were collected. After treatment, all of the 91 patients were followed-up to 2 years. The patients were divided into favorable prognosis group and unfavorable prognosis group according to follow-up results. The differences in all parameters between the two groups werecompared. The relationship between Ang-1, Ang-2 and lung function parameters was analyzed by Pearson correlation analysis. Cox proportional hazard regression model was used to evaluate the effect of clinical parameters on the prognosis of patients with IPF. The effect of Ang-2 in predicting prognosis of patients with IPF was analyzed by receiver operating characteristic (ROC) curve.Results During the 2-year follow-up period, 30 of 91 patients showed a favorable prognosis, and 55 showed an unfavorable prognosis with a poor prognosis rate of 64.71%, and 6 patients withdrew from the study due to loss of follow-up and death. Compared with the favorable prognosis group, Ang-2 level in the unfavorable prognosis group was significantly increased (μg/L: 2.88±1.63 vs. 1.89±1.22,t = 2.909,P= 0.005), but Ang-1 only showed a slight increase (μg/L: 28.70±14.26 vs. 25.62±11.95,t = 1.005,P = 0.318). The results of Pearson correlation analysis showed that Ang-2 level was negatively correlated with forced expiratory volume in 1 second (FVC1) and the percentage of carbon monoxide diffusing capacity accounting for the expected value (DLCO%;r value was -0.227 and -0.206, andP value was 0.147 and 0.253, respectively), but no significant correlation between the level of Ang-1 and FVC1 as well as DLCO% was found (r value was -0.153 and -0.121, andP value was 0.147 and 0.253, respectively). Cox proportional hazard regression model analysis showed that the prognosis of patients with IPF was significantly affected by smoking time and Ang-2 (bothP 0.05). Prognostic analysis showed that the area under ROC curve (AUC) of Ang-2 for predicting prognosis of patients with IPF was 0.692, and the best diagnostic point was 0.35μg/L, the sensitivity was 61.8%, the specificity was 73.3%, the positive predictive value was 69.8%, and the negative predictive value was 65.7% which indicated that Ang-2 could predict the prognosis of patients with IPF.Conclusion Ang-2 could assess the prognosis of patients with IPF, which is expected to be used as an indicator of predicting the prognosis of patients with IPF.
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PURPOSE: To investigate the properties of angiogenin (ANG) as a potential tool for the diagnosis and grading of dry eye syndrome (DES) by analyzing tear protein profiles. METHODS: Tear samples were collected with capillary tubes from 52 DES patients and 29 normal individuals as controls. Tear protein profiles were analyzed with an immunodot blot assay as a screening test. To confirm that the tear ANG levels were in inverse proportion to the disease severity grade, the ANG and lactoferrin (LF) tear contents of normal controls and DES patients were compared in an enzyme-linked immunosorbent assay. RESULTS: In the immunodot blot assay, the ANG area was lower in patients with grades 3 and 4 DES than in normal controls. The areas of basic fibroblast growth factor, transforming growth factor β2, and interleukin 10 were significantly greater than those of normal controls only in grade 4 DES patients, but these proteins were not linearly correlated with dry eye severity. Upon enzyme-linked immunosorbent assay analysis, the mean concentrations of ANG and LF decreased significantly as dry eye severity increased, except between grades 1 and 2. In addition, the ratios of ANG and LF to total tear proteins were correlated significantly with DES severity. CONCLUSIONS: ANG level was significantly lower in DES patients than in normal controls, and was significantly correlated with the worsening severity of DES, except between grades 1 and 2, as was LF. Therefore, ANG may be a useful measure of DES severity through proteomic analysis.
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Angiogenesis Inducing Agents/pharmacology , Dry Eye Syndromes/diagnosis , Enzyme-Linked Immunosorbent Assay , Follow-Up Studies , Immunoblotting , Proteomics/methods , Ribonuclease, Pancreatic/pharmacology , Severity of Illness Index , Tears/chemistryABSTRACT
Neamine, a non-toxic derivative of neomycin, has recently been shown to have antitumor activities in various types of cancers. However, its effect on pancreatic cancer is still unknown. The study aimed to investigate its antitumor activity on pancreatic cancer and the underlying mechanisms. MTT assay was used to observe the effect of neamine on angiogenin (ANG)-induced AsPC-1 cell proliferation. Tissue microassay and immunofluorescence staining were used to detect the expression of ANG and its nuclear translocation, respectively. Tumor xenografts were established by subcutaneous inoculation of AsPC-1 pancreatic cancer cells into the right flanks of nude mice, and neamine was injected subcutaneously. Immunohistochemistry was done to observe the expression of ANG, CD31 and Ki-67 in tumor xenografts. It was found that neamine blocked the nuclear translocation of ANG effectively and inhibited ANG-induced AsPC-1 cell proliferation in a dose-dependent manner. Neamine had anti-tumor effects on AsPC-1 xenograft models. Consistently, neamine reduced the expression levels of ANG, Ki-67 and CD31 in tumor xenografts. It was concluded that neamine may be a promising agent for treatment of pancreatic cancer.
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Adult , Animals , Humans , Male , Mice , Middle Aged , Antibiotics, Antineoplastic , Pharmacology , Therapeutic Uses , Carcinoma , Drug Therapy , Cell Line, Tumor , Cell Proliferation , Framycetin , Pharmacology , Therapeutic Uses , Ki-67 Antigen , Genetics , Metabolism , Mice, Inbred BALB C , Mice, Nude , Pancreatic Neoplasms , Drug Therapy , Platelet Endothelial Cell Adhesion Molecule-1 , Genetics , Metabolism , Ribonuclease, Pancreatic , Genetics , MetabolismABSTRACT
OBJECTIVES: In numerous malignancies, angiogenin (ANG) and Maspin are important proangiogenic and antiangiogenic regulators, respectively. The aim of this study was to identify potential relationships between the biological roles of these two proteins in laryngeal squamous cell carcinoma (LSCC). METHODS: Immunohistochemical staining for ANG and Maspin was performed on specimens from 76 consecutive LSCC patients treated with surgery alone, considering the subcellular pattern of Maspin expression. Univariate and multivariate statistical models were used for prognostic purposes. RESULTS: On univariate analysis, a different level of ANG expression was seen for patients stratified by subcellular Maspin expression pattern: the mean ANG expression was higher in cases with a nonnuclear MASPIN expression than in those with a nuclear pattern (P=0.002). Disease-free survival (DFS; in months) differed significantly when patients were stratified by N stage (P=0.01). Patients whose Maspin expression was nonnuclear (i.e., it was cytoplasmic or there was none) had a significantly higher recurrence rate (P or =5.0% had a significantly shorter DFS than those with an ANG expression or =5.0% was a significant, independent, negative prognostic factor in terms of DFS (P=0.041). CONCLUSION: Our results support the hypothesis that a higher ANG expression is associated with a nonnuclear Maspin expression pattern in patients with LSCC. Further studies are needed to clarify the relationship between the ANG and Maspin pathways, and their potential diagnostic and therapeutic role in LSCC.
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Humans , Carcinoma, Squamous Cell , Cytoplasm , Disease-Free Survival , Intracellular Space , Laryngeal Neoplasms , Models, Statistical , Multivariate Analysis , Prognosis , RecurrenceABSTRACT
Objective: To investigate the effect of autologous angiogenin(ANG)-overexpressing marrow stromal cells transplantation on the neoangiogenesis and cardiac function in a porcine model of chronic ischemia. Methods: Porcine models of chronic ischemia were established and were randomized into the following 3 groups: autologous marrow stromal cells transfected with Ad-ANG (Group I, n=11), autologous marrow stromal cells without transfection (Group II, n=10), and controls injected with serum-free DMEM medium (Group III, n=10). The autologous marrow stromal cells were cultured and were transfected with Ad-ANG, and the cells were labeled by CM-Dil. The cels were transplanted to the ischemia site of porcine heart. The parameters we observed included Rentrop score, ejection fraction, percentage of infracted area, CM-Dil labeled cells under fluorescence microscope, number of blood vessels and expression of ANG protein. Results: Compared with group H and IE, Group I had significantly increased Rentrop score, ejection fraction, numbers of blood vessels in the infarcted areas, CM-DiI labeled cells, and ANG protein expression, but significantly decreased infarcted area (P < 0. 05 or P < 0. 01). Conclusion: Transplantation of autologous angiogenin-overexpressing marrow stromal cells can yield a high survival rate of cells, greatly improving the blood supply of the infarcted cardiac area and cardiac function in porcine model.
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Retinopathy of premature is characterized by retinal vascular dysplasia,angiogenesis,fiber proliferation and retinal detachment,which can lead to a variety of serious complications,including permanent blindness.Angiopoietin-1,which has a close relationship with the incidence of ROP,is an important factor affecting angiogenesis.This paper will discuss the relations between angiopoietin-1 and retinopathy of premature.
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BACKGROUND:Shengji Yuhong col agen showed good curative effect of promoting angiogenesis and tissue healing compared with Shengji Yuhong Gao and col agen alone or gelatin alone. OBJECTIVE:To explore the curative effect and mechanism of subcutaneous implantation of Shengji Yuhong col agen in rabbits in promoting angiogenesis and repair. METHODS:Shengji Yuhong col agen as the experimental group and collagen as the control group was implanted inside the rabbit subcutaneous pockets of the back of New Zealand rabbits. The implanted samples and surrounding tissues were obtained at 3, 7, 14, 28 and 56 days fol owing surgery. Pathological sections were made and the repair of surrounding tissue was observed. Hemoglobin levels in col agen were measured. Immunofluorescence and CD34 dyeing marking method were utilized to observe capil ary angiogenesis. Western blot assay was employed to examine vascular endothelial growth factor and angiogenin-1 expression. Immunohistochemistry was used to observe the secretion of typeⅠ and Ⅲ col agen on the surrounding tissues. RESULTS AND CONCLUSION:The experimental group showed increased subcutaneous vascularization. There were reduced inflammatory exudation, granulation tissue hyperplasia, and mature fiber connective tissue at 28 days. Angiogenesis and hemoglobin contents were greater in the experimental group than in the control group (Pidentical between the experimental and control groups. However, the secretion of type Ⅲ col agen was higher in the experimental group than in the control group at 28 and 56 days (Pcol agen was lower in the experimental group than in the control group at 28 and 56 days (Pmechanisms of adjusting the protein expression of vascular endothelial growth factor and angiogenin-1. At the same time, it has the function of regulating col agen formation with better ratio of typeⅠ and type Ⅲ col agen to acquire higher quality of wound healing with reduced scar formation.
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Objective To observe the levels of Ang - 1 and NF-κB in lung tissue and to aseess the severity of ALI induced by phosgene in order to clarify the mechanism of the protective effect of Ang - 1 on phosgene induced ALI.Method Rats were randomly divided into phosgene group and air group.Another rats were randomly (random number) divided into phosgene group,phosgene + PDTC group and air group.Lung tissue was collected to weigh and calculate the wet / dry weight ratio,measure BALF,white blood cell count,total protein and Ang-1 at given time after exposure to phosgene/air and PDTC.The Ang - 1 and NF-κB levels in lung tissue were measured with Western blot and immunohistochemistry.Data were analyzed by using SPSS 16.0 statistical package and comparisons between groups were carried out byusing One-Way ANOVA analysis and LSD -t test,α ＜ 0.05.Results Serum angiopoietin -1 level became lesser within 48 hours after exposure to Phosgene.The severity of ALI became worser with time elapsing.Ccompare with air group,the severity of ALI in phosgene group was worser with time elapsing ( P ＜ 0.05).Compared with phosgene + PDTC group,the serum angiopoietin -1 and arterial oxygen partial pressure in phosgene group were lower ( P ＜ 0.05).The severity of ALI of rats in phosgene group were worser than that in phosgene + PDTC group ( P ＜ 0.05).Serum angiopoietin -1 and partial pressure of oxygen of rats in phosgene group were higher than those in phosgene + PDTC group ( P ＜ 0.05).Immunohistochemistry test showed that the expression of Ang-1 in lung tissue in air group were normal,and Ang-1 in phosgene group were significantly reduced,and Ang-1 in PDTC intervention group was higher than that in phosgene group and lower than that in air group.The above results were confirmed by Western blot test which was consistent with the results of immunohistochemistry test.Similarly,the levels of NF-κB in lung tissue determined by using both Western - blot and immunohistochemistry were consistant,and results of both methods showed that the expression of NF - κB in air group was normal,and it increased in phosgene group,and the expression of NF-κB in phosgene + PDTC group was lower than that in phosgene group.Conclusions The serum level of Ang-1 was decreasing within 48 hours after ALI.Ang-1 was negatively correlated with the sevfity of phosgene induced ALI.Ang-1 likely had an effect on NF-κB signaling pathway,ameliorating the inflammation mediated by cytokines,reducing lung endothelial permeability and in turn lessening the severity of ALI.
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Objective: To inhibit the expression of angiogenin (ANG) by recombinant adeno-associated virus (AAV) mediated RNA interference (RNAi) and to observe its influence on the growth of human lung squamous cancer cells line, SK-MES-1. Methods: A recombinant adeno-associated virus vector was constructed and used to deliver small hairpin RNA (shRNA) targeting ANG into SK-MES-1 cells; SK-MES-1 cells and SK-MES-1 cells transfected with AAV-Null were taken as control. The inhibitory effect of adeno-associated virus mediated RNAi on ANG expression and subsequent influence on the growth, tumorigenesis, and microvessel density of SK-MES-1 tumors were also evaluated. Results: In vitro experiment showed that AAV-shANG was successfully constructed. Western blotting revealed that AAV-shANG effectively infected SK-MES-1 cells and the expression of ANG was obviously decreased, 72 h after transduction. In vivo study showed that, compared with the other 2 groups, the suppression of ANG in AAV-shANG group significantly inhibited the growth of SK-MES-1 cells and decreased the MVP in nude mice (P<0. 05). Conclusion: The constructed AAV-shANG can effectively inhibit ANG protein expression in SK-MES-1 cells, which paves a way for the therapy of human lung squamou cancer.
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Angiogenin, a potent inducer of angiogenesis, is expressed in human endometrium. This study was performed to compare the expression of angiogenin mRNA level in the eutopic endometrium from women with and without endometriosis. Thirty-two women with advanced stage endometriosis and 29 control women were recruited. Following isolation of total RNA from endometrial tissue and reverse transcription, cDNA samples were amplified by real time polymerase chain reaction to quantify the expression of angiogenin genes. In selected patients, immunohistochemical staining was utilized to localize the area of angiogenin expression. Angiogenin mRNA level was significantly lower in the endometriosis group than in the control group during the secretory phase, especially the mid-secretory phase, and the decline was observed mainly in the women who presented with infertility. Within the endometriosis group, angiogenin mRNA levels did not differ between the proliferative and secretory phases, but, in the control group, the level in the secretory phase was higher than that during the proliferative phase. Immunohistochemistry showed that the glandular epithelial cell layer was decorated positively in both groups. These findings suggest that the relative deficiency of angiogenin expression in the secretory endometrium could impair implantation in women with advanced stage endometriosis.
Subject(s)
Adult , Female , Humans , Endometriosis/metabolism , Endometrium/metabolism , Fertility , Gene Expression Regulation , Immunohistochemistry/methods , Menstrual Cycle , Models, Biological , RNA, Messenger/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Ribonuclease, Pancreatic/biosynthesisABSTRACT
Objective:The purpose is to observe the change of the number of apoptosis cell in the reversible-ischemia rat brain after the intraperitoneal injection of rhANG. Methods:With the use of a reversible(middle cerebral artery occlusion (MCAO))m odel in rats,apoptosis cells were assessed by(terminal deoxynucleotedyl transferase(TdT))-mediated dUTP-biotin nick end labe ling(TUNEL) histochemical methods in sham,brain ischemia control and rhANG intraperitoneal injection groups.Results:There was no significant difference in the number of the apoptosis cell in the penumbra zone in the ischemic brain between the rhANG intraperitoneal injection group and the brain ischemia control group.The number of apoptosis cell in the ischemic lateral brain was increased significantly compared with the non-ischemia lateral in the brain ischemia control group and the rhANG intraperitoneal injection group. Conclusion:There was no significant difference in the number of the apoptosis cell in the penu mbra zone in the ischemic rat brain after the intraperitoneal injection of rhANG(15ng/100g)compared with the control group.
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To develop a novel therapeutic angiogenesis for the treatment of cardiovascular diseases, angiogenin (ANG1) was examined as a potential therapeutic gene. An adeno-associated virus (AAV)-mediated gene delivery system was used to measure the therapeutic efficacy of ANG1. Using a triple co-transfection technique, rAAV-ANG1-GFP, rAAV- VEGF-GFP and rAAV-GFP vectors were produced, which were then used to infect human umbilical vein endothelial cells (HUVECs) in order to evaluate in vitro angiogenic activities. Their protein expressions, tagged with green fluorescent protein (GFP), were monitored by confocal microscopy. The functional activities were measured using wound-healing HUVEC migration assays. The number of migrated cells stimulated by both the expressed ANG1 and the VEGF in rAAV-infected HUVECs increased almost twice the number observed in the expressed GFP control. In vivo angiogenic activities of the expressed ANG1 or VEGF were determined using mouse angiogenesis assays. The angiogenic activities of ANG1 or VEGF expressed in the injected mice were increased by 1.36 and 2.16 times, respectively, compared to those of the expressed GFP control. These results demonstrate that the expressed ANG1 derived from rAAV infection has in vitro and in vivo angiogenic activities and suggest that the rAAV-ANG1 vector is a potential strategy for therapeutic angiogenesis.
Subject(s)
Animals , Humans , Male , Mice , Cell Movement , Cells, Cultured , Dependovirus/genetics , Endothelial Cells/metabolism , Gene Transfer Techniques , Genetic Vectors , Mice, Inbred C57BL , Neovascularization, Physiologic , Ribonuclease, Pancreatic/biosynthesis , Umbilical Veins/cytology , Vascular Endothelial Growth Factor A/biosynthesisABSTRACT
0.05).In group I,the mRNA levels of Ang-1 were lower than those in group C during 3-6 h after onset of ischemia(P
ABSTRACT
PURPOSE: The purpose of this study was to verify that the expressions of angiogenin, transforming growth factor-beta(TGF-beta), vascular endothelial growth factor(VEGF), human apurinic/apyrimidinic endonuclease(APEX) and tumor necrosis factoralpha(TNF-alpha) were associated with the tumorigenesis of the oral squamous cell carcinoma(OSCC). MATERIALS AND METHODS: Fifty-one samples of OSCC and fifteen normal oral mucosae were obtained to analyze the expression levels of above five factors. mRNA expressions were quantified by the quantitative competitive PCR(QC-PCR) method. After 2% agarose gel electrophoresis stained with ethidium bromide, the concentration of mRNA was calculated by a digital image analysis system. The expression levels of angiogenin, TGF-beta, VEGF, APEX and TNF-alpha were compared by unpaired Student's ttests between cancer and normal tissues. We analyzed statistically to find the cut-off values that would be useful as diagnostic markers, and the linear regression analysis between every two factors of these five factors by SAS system. RESULTS: All of these five factors (angiogenin: P<0.0037, TGF-beta: P<0.0001, VEGF: P<0.0102, APEX: P<0.0023, TNF-alpha: P<0.0074) were significantly correlated with OSCC. In the analysis to find the cut-off values for the diagnosis, we could not find any value that had a reasonable sensitivity and specificity. In the linear regression analysis, there were correlations between angiogenin and TNF-alpha, TGF-beta and VEGF, TGF-beta and APEX, TGF-beta and TNF-alpha, VEGF and APEX, VEGF and TNF-alpha, APEX and TNF-alpha. CONCLUSION: Our results suggest that not only angiogenin, TGF-beta, VEGF, APEX and TNF-alpha are significantly associated with the tumorigenesis, but also the close relationship between these factors might enhance the tumorigenesis of OSCC. We can not find clinical availability for diagnosis.