Dysfunction of endothelial progentior cells in the angiographically defined coronary atherosclerosis / Монголын Анагаах Ухаан

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)

Effect of STAT5 Pathway Inhibitor Pimozide on NO and iNOS Expressions in the Inflammation Model of RAW264.7 Cell / 中国药房

OBJECTIVE:To investigate the effect of STAT5 pathway inhibitor pimozide on the expressions of nitric oxide (NO)and nitric oxide synthetase(iNOS)in the model of mouse macrophage RAW264.7 inflammation induced by lipopolysaccha-ride (LPS). METHODS:RAW264.7 cells in logarithmic growth phase were divided into blank control group,drug control group (10μmol/L pimozide),model group(1μg/ml LPS)and the pimozide groups of low,middle and high doses(2.5,5 and 10μmol/L), where the corresponding cells were given pimozide 30 min before the administration of LPS,and then were cultured for 24 h. Griess method was used to determine the content of NO in the supernate of cell culture solutions of all groups,real-time quantita-tive polymerase chain reaction(RT-PCR)to determine iNOS mRNA expression,and Western blot method to determine the protein expression of iNOS and phosphorylated STAT5(p-STAT5). RESULTS:The content of NO,iNOS mRNA and protein expressions and the content of p-STAT5/STAT5 in the cells in the model group were higher than those in the blank control group,with statisti-cally difference (P<0.01). Compared to the model group,the pimozide groups of middle and high doses had lower content of NO,iNOS mRNA and protein expressions and the content of p-STAT5/STAT5 in the cells,with statistically difference(P<0.01 or P<0.05). CONCLUSIONS:STAT5 pathway inhibitor pimozide can inhibit the release of NO by inhibiting iNOS mRNA and pro-tein expressions in cells.

An experimental study on inhibitory effects and mechanisms of N-butanol extract of Potentilla anserine L. against hypoxia-induced nitric oxide on hippocampus neurons of rats / 中国中西医结合急救杂志

Objective To study in vitro the inhibitory effects and mechanisms of N-butanol extract of Potentilla anserine L.(NP)against hypoxia-induced nitric oxide(NO)in hippocampus neuron of rats. Methods The models of hippocampus neurons hypoxia injury of Sprague-Dawley(SD)neonatal rats were cultured in vitro. The cultured hippocampus neurons were divided randomly into blank control group, hypoxia injury model group, nimodipine group(2 μmol/L)and NP high(250.0 mg/L),middle(62.5 mg/L),low(15.6 mg/L)dose groups. The activities of hippocampus neurons were examined by methyl thiazolyl tetrazolium(MTT)assay,and meanwhile their contents of nitrogen monoxidum(NO)were detected. Half quantity reverse transcription-polymerase chain reaction(RT-PCR)and Western blotting were used to detect neuronal nitric oxide synthetase(nNOS)mRNA and protein expression levels respectively in each group,immunocytochemistry stain was used to detect protein positive rate. Results Compared with blank control group,the activity of neuron〔absorbance(A)value〕was significantly decreased(0.0826±0.0095 vs. 0.3315±0.0105),content of NO(μmol/g:0.0509±0.0027 vs. 0.0291±0.0032), the expression levels of nNOS mRNA (0.1463±0.0081 vs. 0.0801±0.0058), the positive rate of nNOS〔(74.4238±3.9423)%vs.(28.3714±4.1361)%〕,the expression levels of nNOS protein(A value:1.9130±0.0471 vs. 0.5068±0.0368)were all significantly increased in the hypoxia injury model group(all P0.05). Conclusions NP can ameliorate the injury of rat hippocampus neurons induced by hypoxia in vitro. The possible mechanisms might be related to the effective inhibition of the synthesis of nNOS and NO excessive generation.

Prevention and treatment of nitric oxide synthetase inhibitor on galactose cataract in rat / 中华实验眼科杂志

Background Researches showed that the content of nitric oxide (NO) and nitric oxide synthetase (NOS) increases in blood,aqueous humor and tear of cataract patient.But the function of NO and NOS in cataract formation is still elusive.Objective The aim of this study was to explore the prevention and treatment effect of NOS inhibitor,1-nitro-arginine methyl ester (L-NAME),on galactose cataract.Methods Sixty clean three-week-old Wistar rats were equally and randomly divided into 3 groups.0.9％ Normal saline solution (30 ml/kg) was subcutaneously injected every day for 30 days in the rats of the control group,and 50％ of D-galactose solution (30 ml/kg) was used in the rats of the model and L-NAME group at the same way.L-NAME eye drops was simultaneously administered in the L-NAME group 3 times per day for 30 days.The eyes of the rats were examined under the slit lamp in 10,20 and 30 days,and the degree of lens opacification was scored.Lenses of the rats were obtained at the end of this experiment for the detect of NO,NOS contents.Flow cytometry was used to assay the caspase-3 level of rat lens.Repeated measurement two factor analysis of variance was used to analyze the difference of lens opacification scores in different groups and different time points,and one-way ANOVA was used to analyze the differences of NO,NOS and caspase-3 contents in lens among the groups.Results Lens opacification appeared in 10 days after injection of 50％ D-galactose solution in the rats of the model group and L-NAME group.Lens opacification score was higher among the different groups and different time points (Ftime =435.251,P =0.000 ;Fgroup =395.120,P=0.000).NO content in the lens was (0.45±0.15) μmol/g,(2.67 ± 0.47) μmol/g and (1.68±0.34) μmol/g in the control group,model group and L-NAME group,showing a significant difference (F=58.872,P=0.000).The NOS contents in the lens was (0.0160±0.0020) U/ml,(0.0370±0.0040) U/ml and (0.0270±0.0010) U/ml in the control group,model group and L-NAME group,showing a significant difference (F =66.174,P=0.000).Caspase-3 contents in the lens was (339.4 ± 37.9),(697.7 ± 46.5) and (650.7 ± 53.1),Showing a significant difference among them (F =100.005,P =0.000).Conclusions The increase of NO,NOS and caspase3 levels are associated with lens opacification.Topical administration of L-NAME eye drops can down-regulate NOS content in lens,reduce the NO formation and inhibit the apoptosis of lens epithelial cells.

Measurement of nitric oxide levels and activity of nitric oxide synthetase in the patients with depression / 中国综合临床

Objective To measure the correlation between the levels of blood serum nitric oxide(NO) ,the activity of nitric oxide synthetase(NOS) and depression.Methods 136 patients with depression were diagnosed by CCMD-3 and 120 healthy control subjects were included in the study.NO and NOS levels in blood serum of both groups were tested, and the variation of NO and NOS between the two groups were compared.Results The blood se-rum NO levels[ (70.05±10.34)μmol/L w.(67.17±16.52) μmol/L] and the activity of NOS [(29.49±5.12)U/L vs.(26.99±2.87) U/L] in the patients with depression were significantly higher than those in the control group(P<0.05).The blood serum NO levels and the activity NOS in the treatment patients was lower than the oth-er patients[(74.42±8.80) μmol/L vs.(78.81±12.28) μmol/L;(27.71±5.46)U/L vs.( 30.49±4.65 )U/L,P <0.05].Conclusion The blood serum NO levels and the activity of NOS increase in the patients with de-pression.NO might be involved in the pathogenesis of depression.The antidepressants might descend the NO levels so to relieving depression.

Efficacy of mIL-18BP and mIL-4 adenoviral coexpression gene therapy for cyclooxygenase-2 and inducible nitric oxide synthetase in murine collagen-induced arthritis / 中华风湿病学杂志

Objective A recombinant adenoviral vector containing mIL-18BP and mIL-4 fusion gene(AdmIL-18BP/mIL-4) was constructed and used to investigate the role of mIL-18BP and mIL-4 in medula-ring the expressions of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthetase (iNOS) and their inducing products PGE2, NO in murine collagen-induced arthritis. Methods Male DBA-1/BOM mice were used in this study. Mice with CIA were intra-articularly injected with 107 pfu/6μl of AdmIL-18BP/mIL-4.Intra-articular injections of AdLacZ or PBS were used as controls. The mRNA expression of COX-2, iNOS in synovial tissue was analyzed by semi-quantitative RT-PCR. Expression of COX-2 and iNOS protein was estimated by Western blot method. The production of PGE2 and NO in synovia was detected by competitive ELISA and enzyme reduction of nitrate. Results The expression of COX-2, iNOS mRNA in routine synovial tissue of AdmIL-18BP/mIL-4 treatment group was significantly lower than that of AdLacZ group (0.15 vs 0.42,P<0.01 ; 0.05 vs 0.77, P<0.01) and PBS group (0.15 vs 0.65, P<0.01; 0.05 vs 0.64, P<0.01 ). And the protein expression of COX-2, iNOS from AdmIL-18BP/mIL-4 treatment group was also obviously lower than that of AdLacZ group (0.08 vs 0.92, P<0.01; 0.11 vs 1.00, P<0.01) and PBS group (0.08 vs 0.77, P<0.01; 0.11 vs 0.84, P<0.01 ). The PGE2 and NO production in synovia of AdmIL-18BP/mIL-4 treatment group was significantly lower than that of AdLacZ group [(0.68x0.06) vs (2.58±0.21)ng/mL, P<0.01; (23.4+2.5) vs (60.0±11.3)μmol/L, P<0.01 ] and PBS group [(0.68±0.06) vs (2.57±0.20)ng/mL, P<0.01; (23.4+2.5) vs (60.3±13.4)μmol/L, P<0.01]. Conclusion These data indicat that local over-expre-ssion of mIL-18BP and mIL-4 can down-regulate COX-2, iNOS and their induced product PGE2, NO in CIA mice. The combination treatment with mIL-18BP and mIL-4 is a promising therapeutic target for RA.

Escenciales en biomedicina (biología, patobiología y bioclínica) humana de las NOSS (Óxido Nítrico-Sintetasas): [revisión] / Essentials in human biomedicine (biology, patobiology and bioclinic)of the nitric oxide synthetases (NOS): [review]

El Óxido Nítrico (NO) es un radical libre gaseoso que juega roles prominentes en señalamiento celular, expresión y regulación génica, energética celular, proliferación celular y citostasis, e inmunidad celular y tolerancia, incluyendo funciones inflamatorias. Aunque NO sirve a roles beneficiosos como citotrófico, vasodilatador, antiangiogénico, anti-trombótico, anti-inflamatorio, defensa inmune del huésped, antiproliferativo y antioxidante, su excesiva producción puede ser citotóxica, vasoconstrictora, pro-angiogénica, protrombótica, pro-inflamatoria, proproliferativa y oxidante, a causa de la endógena producción de intermediarios reactivos del nitrógeno y/o el oxígeno. Esta revisión explora el conocimiento colectivo del rol de las NO-Sintetasas (NOSs) en biología, patobiología, bioclínica humana y nuevas oportunidades en prevención y tratamiento...

The Nitric Oxide (NO) is a gaseosus free radical that plays prominent roles in cell signaling, gene expression and regulation, cellular energetics, cell proliferation and cytostasis, and cell immunity and tolerence including inflammatory functions. Althoug NO serves benificial rolesas cytotrophic, vasodilator, anti-angiogenic, antithrombotic, anti-inflammatory, host defense, antiproliferativeand antioxidant, excessive production can be cytotoxic, vasoconstrictor, pro-angiogenic, prothrombotic, proinflammatory, pro-proliferative and oxidant, because endogenous production of reactive nitrogen and/o oxygen intermediates. The review explores the collective knowledge of the role of NO-Synthetases (NOSs) in human biology, pathobiology, bioclinic and new opportunities in prevention and treatment.

Effects of nitric oxide on experimental hepatic fibrosis in rats / 中国药理学通报

AIM: To study the role of nitric oxide (NO) in the pathogenesis of hepatic fibrosis. METHODS: To establish the model of hepatic fibrosis in rats and treated with L-Arg (preNO) and L-NNA (NOS inhibitor). The degree of hepatic fibrosis, the levels of HA, AST, ALT determined with histology, radioimmunoassay, autochemol-analysis, respectively. NO and NOS were also determined. RESULTS: The degree of hepatic fibrosis, the levels of AST, ALT and HA in L-Arg group were all lower than those in hepatic fibrosis control group. While the levels of HA and AST in L-NNA group were higher than that in hepatic fibrosis control group. CONCLUSION: NO has the function of protecting liver cells and anti-hepatofibrosis liver fibrosis induced by CCl4 in rats.

Influence of Aspirin on growth and NOS expression in astrocytoma cell line / 中国免疫学杂志

Objective:To investigate the effect of Aspirin on the proliferation and NOS expression in astrocytoma cell line,and probe into ins mechanism.Methods:The effect of Aspirin on the growth of astrocytoma cells evaluated by MTT assay;NOS protein levels were determined by immunohistochemistry,NO and CEA concentration in the medium were determined by Griess assay and lepton catch immuning method respectively.Results:Aspirin can inhibit the growth of astrocytoma cells,induce the expression of iNOS,increase the concentration of NO in the medium. The effect of these were in a concentration dependent pattern. Moreover,Aspirin can reduce the concentration of CEA in the medium.Conclusion:Aspirin inhibit the growth of astrocytoma cell line.UP regulated iNOS expression resulting a increase of NO concentration are ascribed to mechanism of antrproliferation activity of Aspirin. CEA is a good indicator in monitoring curative effect of astrocytoma.

The expression of nitric oxide synthetase in the experimental tooth movement in rats / 대한치과교정학회지

Nitric oxide(NO) has been reported to be one of the mediators relating to bone remodelling. Nitric oxide is synthesized from L-arguinine by nitric oxide synthetase(NOS), which is largely divided into two groups. One group which is composed of NOS1 and NOS3, is dependent of calcium or calmodulin. The other consisted of NOS2, which is independent of calcium or calmodulin. NOS is thought to be a possible intermediate affecting in the course of tooth movement. This study was designed to evaluate the expression of nitrous oxide synthetase(NOS) in periodontal tissue during the experimental movement of rat incisors, by LSAB(labelled streptavidine biotin) immunohistochemical staining for NOS2 and NOS3. Twenty seven Sprague-Dawley rats were divided into a control group(3 rats), and 6 experimental groups(24 rats), to which 75g of force was applied, with helical springs across the maxillary incisors. Rats of experimental groups were sacrificed at 12 hours, 1, 4, 7, 14 and 28 days after force application, respectively. After that, the tissues of the control group and experimental groups were studied immunohistochemically. The results were as follows: 1. In control group, the expression of NOS3 was rare in gingiva, dentin, periodontal ligament and alveolar bone, and was mild in the capillaries of pulp and intermaxillary suture. And the expression of NOS2 showed similar pattern to that of NOS3. 2. There were no differences in the expression of NOS2 or NOS3 in dentin, gingiva, cementum, cementoblast and odontoblast, between control and experimental groups, regardless of the duration of the force application. 3. The expression of NOS3 began to increase at 4 days and showed to the highest degree at 7 days after force application, in the apical region of pressure side of periodontal ligament in experimental groups. 4. The expression of NOS3 in alveolar bone was rare until 7 days, after which it increased to mild degree at 14 days through 28 days in experimental group. But there was no difference between pressure and tension side of periodontal ligament. 5. The expression of NOS2 in periodontal ligament was mild from 7 days after force application, regardless of the side of periodontium, which was generally more evident than that of NOS3 . 6. The expression of NOS2 in alveolar bone increased to mild degree at 14 days after force application, and it was evident in osteoblasts, osteoclasts and osteocytes. And the expression of NOS2 was little more stronger in the tension side than that of pressure side of alveolar bone.

A Study on Relationship between iNOS and p53 Expression in Human Prostatic Carcinoma / 中国医师杂志

Objective To investigate the association of inducible nitric oxide synthetase (iNOS) expression with the expression of mutation type p53 (mt p53) in prostatic carcinoma (PCa). Methods The expression levels of iNOS and mt p53 were in 28 PCa specimens and 10 cases of benign prostatic hyperplasia (BPH) were detected by immunohistochemical method. Results The expression levels of both iNOS and mt p53 in PCa were higher than those in BPH. The expression of iNOS was related to the expression of p53 protein. Conclusion iNOS and mt p53 may play a role in the pathogenesis of PCa.

Comparison Between Borneol-induced Opening of Blood-brain Barrier and Its Pathological Opening / 中药新药与临床药理

Objective: To observe the difference between borneol-induced opening of blood-brain barrier (BBB) and its pathological opening . Methods: The expression of inducible nitric oxide synthetase (iNOS) in brain microvascular endothelial cells in normal rats and brain-injury rats before and after treatment of borneol was examined by immunohistochemical methods with S-P staining and iNOS antibody. Results: iNOS expression was not different in normal rats before and after treatment. However, iNOS expression was increased in brain-injury rats and borneol could inhibit the expression of iNOS. Conclusion: Borneol-induced opening of BBB is different from pathological opening of BBB. Borneol can increase the pathological opening of BBB and has a protective effect on brain tissue.

Effects of Nocodazole on Fine Structures in Co-cultured Vascular Endothelial Cells and Cancer Cell Lines / 대한해부학회지

Endothelial cells were isolated from the aortic intima of Sprague-Dawley species, rat. These cells and each cancer cell line (HeLa, Hep G2, A549, L929 and NIH/3T3 cells) were co-cultivated in alpha-MEM with 3 micrometer or 30 micrometer nocodazole. To investigate the influences induced by nocodazole, the morphological changes were observed under inverted microscope and transmission electron microscope, the amounts of fibronectin produced by vascular endothelial cells and cancer cell lines and the activities of nitric oxide synthetase synthesized mainly by endothelial cells were analyzed in the aspects associated with fine structural changes. The vascular endothelial cells of control group at the 1st, 2nd and 3rd days extended the cell processes, which contacted with cells from all cell lines investigated, but the endothelial cells of nocodazole-treated groups didn't possess the processes. All cell lines in nocodazole-treated groups had a large number of micronucleated cells, but endothelial cells didn't show micronuclei. Compared with control group, the endothelial cells of nocodazole-treated groups at the 1st, 2nd and 3rd days showed the decrease of amounts of fibronectin because of the increase of heterochromatin area. The amounts of fibronectin increased in all cell lines of nocodazole-treated groups at the 2nd and 3rd days whereas the nuclear folding or the dilatation/numerical increase of rough endoplasmic reticulum didn't appear. The activities of nitric oxide synthetase heightened in endothelial cells of nocodazole-treated groups, and therefore the considerable changes in fine structures such as vesicles, lysosomes, liposomes, pyknosis and cell lysis occurred even though the extent of changes differed among the cell lines. Taken together, the materials such as fibronectin or nitric oxide synthetase produced by endothelial cells directly or indirectly acted on cancer cells, and the amounts of fibronectin and the vesicles, lysosomes, liposomes and cell lysis seemed to be much more increased or enforced. Therefore, co-culture system seemed to work better for the investigation of actions of nocodazole and the role of endothelial cells in cancer cells research. Also, the co-culture system was closer to the in vivo state and more favorable in studies for proliferation or metastasis of cancer cells.

Endothelial cells from the rat aorta in culture / 대한해부학회지

Endothelial cells were isolated from the intima of Sprague-Dawley rat aorta with 0.2% collagenase solution and cultured in MCDB 131 medium. All endothelial cells regardless of passages and the freezing-thawing were cultured in M-199, alpha-MEM, and MCDB 131 media. Thereafter, the morphological findings of the cells were observed under light and electron microscopes. The activities of nitric oxide synthetase of endothelial cells were investigated with NADPH-diphorase staining. Production of von Willebrand factor, cytoskeletal proteins, and extracellular matrix proteins in endothelial cells was identified with PAP. By treating with 0.2% collagenase solution for 30 minutes, and then incubating the cells in MCDB 131 medium for 3 days, only endothelial cells could be separated. Endothelial cells formed a monolayer with typical cobblestone pattern (polygonal-shaped morphology) 7-10 days after seeding. The growth patterns of endothelial cells were similar regardless of their cultured states (primary cultured cells, subcultured or thawed). The existence of nitric oxide synthetase, von Willebrand factor, beta-actin, fibronectin, and laminin in endothelial cells was confirmed. However, above-mentioned materials were quantitatively less in cells grown with MCDB 131 medium compared to those cultured with M-199 and alpha-MEM. Weibel-Palade bodies were observed by transmission electron microscopy. Cultured cells in MCDB 131 medium compared to those cultured with M-199 and alpha-MEM were relatively fewer in number of rough endoplasmic reticulum, mitochondria, free ribosome, vesicle, and liposome and especially showed the absence of basement membrane. Taken together, the cultivation of endothelial cells from the intima of rat aorta was possible. Endothelial cells in vitro synthesized various materials such as von Willebrand factor etc. and had characteristic organelles (Weibel-Palade body etc.). It suggested that it would be more advantageous to culture the endothelial cells in M-199 with 20% fetal bovine serum and alpha-MEM with 10% bovine calf serum rather than in MCDB 131 endothelial cell culture medium only with 0.7% dialyzed serum to maintain characteristics of endothelial cells in vivo

Effects of Tianshu capsule on plasma NO,NOS,CGRP contents and hemodynamics of migraine animal models / 临床神经病学杂志

Objective To investigate the effects of Tianshu capsule on vasoactive substances in blood plasma or brain and hemodynamics in migraine animal models. Methods After subcutaneously administration of nitroglycerin (NTG) and tube feeding Tianshu capsule, the radioimmunoassay or spectrophotometry was used to measure the plasma contents of calcitonin gene related peptide (CGRP), nitric oxide (NO) and nitric oxide synthase (NOS) in rats.The distribution of NOS1, CGRP in rat nucleus tratus spinalis nervi trigemini was detected by immunohistostaining, and the changes of internal carotid artery flow velocity in rabbit were measured by transcranial Doppler.Results The levels of NO, NOS and CGRP were elevated in the plasma of migraine rats. Tianshu capsule could inhibit these increases, and the moderate-and high-dose had much significant effects (P

Influence of aspirin on growth and NOS expression in astrocytoma cell line / 中国药理学通报

AIM To investigate the effect of aspirin on the proliferation and NOS expression in astrocytoma cell line, and probe into its mechanism. METHOD The effect of aspirin on the growth of astrocytoma cells was evaluated by MTT assay; NOS protein levels were determined by immunocytochemistry, NO and CEA concentration in the medium were determined by Griess assay and lepton catch immunising method respectively. RESULTS Aspirin inhibited the growth of astrocytoma cells, induced the expression of iNOS, increased the concentration of NO in the medium. The effects of these were centratoin dependent. Moreover, aspirin reduced the concentration of CEA in the medium. CONCLUSION Aspirin inhibits the growth of astrocytoma cell line. Up regulated iNOS expression resulting a increase of NO concentration are ascribed to mechanism of antiproliferation activity of aspirin. CEA is a good indicator in monitoring curative effect of astrocytoma.