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Objective To explore the effects of ischemia and ischemia reperfusion on the proliferation,apoptosis,migration ability,inducible nitric oxide synthase (iNOS) and endothelial nitric oxide synthase (eNOS) expression of endothelial progenitor cells (EPCs).Methods Collection of peripheral blood from volunteers and culture of endothelial progenitor cells in vitro.The cells were divided into three groups:control group,hypoxia group and hypoxia reoxygenation group.Methyl thiazolyl tetrazolium (MTT) assay was used to detect cell proliferation.Transwell chamber method was used to detect cell migration.Cell apoptosis was detected by flow cytometry.Western blot was used to detect iNOS and eNOS expressions.Results A confocal microscope was used to observe the basic adherence of the cells to the wall for about 3 days,and the area became larger.After 7 d of single nucleus cell culture,the growth of colony-like pattern was more than that of spindle.The cell counts of the three groups in the microscope were (1.83 ± 0.92),(5.07± 0.84),(2.11 ± 0.74).Compared with the control group (0.24 ± 0.04),the hypoxia group (0.62± 0.06) could promote EPCs proliferation,and the difference was statistically significant (t =12.142,P ＜ 0.05);While there was no significant difference between the hypoxia reoxygenation group (0.39 ± 0.06) and the control group (P ＞ 0.05).The number of cell migration in the hypoxia group (18.28 ± 2.05) and hypoxic complex oxygen group (14.08 ± 2.11) was not statistically significant compared with the control group (15.14 ± 1.25) (P ＞ 0.05).The apoptosis rate in hypoxia group (34.57 ±0.42)％ and hypoxia reoxygenation group (41.08 ± 0.44)％ was significantly higher than that in control group (24.83 ± 0.38) ％ (x2 =13.427,15.084,P ＜ 0.05).The apoptosis rate of hypoxia reoxygenation group was significantly higher than that of hypoxia group (x2 =9.657,P ＜ 0.05).The expression of iNOS in hypoxia group and hypoxia reoxygenation group was significantly higher than that in control group,and the difference was statistically significant (P ＜ 0.05).Conclusions Ischemia could promote the proliferation of EPCs,and increase the expression of iNOS,but the expression of EPCs was down-regulated after reperfusion.
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Objective To investigate the effect of thyroid stimulating hormone (TSH) on the expression of endothelial nitric oxide synthase (eNOS) and its mechanism in human microvascular endothelial cells (HMEC-1) in vitro culture.Methods Different concentrations of TSH (0,10,50 mIU/ml) were used to intervene HMEC-1.The expression of eNOS mRNA was detected with quantitative polymerase chain reaction (qPCR) method.The protein expressions of eNOS,phosphorylated protein kinase B (p-AKT),protein kinase B (AKT),phosphorylated extracellular signal-regulated kinase (P-ERK),and extracellular signal-regulated kinase (ERK) were determined with Western Blot.Results (1) The expression level of eNOS was significantly decreased by TSH in dose-dependent manner (P ＜ 0.05).(2) TSH could promote the phosphorylation of AKT and ERK (P ＜ 0.05).Conclusions Thyroid-stimulating hormone may inhibit the expression of nitric oxide synthase in human microvascular endothelial cells by activating AKT and ERK signaling pathways.
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Objective To investigate effects of ischemic postconditioning on the nitric oxide ( NO) and nitric oxide synthase ( NOS) in diabetic rat brain tissues .Methods Thirty Wistar rats were diabetic models induced by intraperitoneal injuction of stepto-zotocin (STZ), and randomly divided into three groups: Control group (normal, diabetic), cerebral ischemia group, and ischemic postconditioning ( I-POST) group.The rats of cerebral ischemia group and ischemic postconditioning group were made model of cere -bral ischemia by ligation carotid artery .Hematoxylin-eosin ( HE) was used to observe their pathological changes in control and diabetic groups.Enzyme-linked immunosorbent assay ( ELISA) method was used to detect the expression and changes of NO and NOS in the sera in each group .Western Blot method was used to investigate the expression and changes of NOS in the retinal tissues in each group .Results For I-POST group , brain tissue defects were decreased , neuronal cells were increased , serum inducible NOS ( iNOS) content was significantly lower than endothelial NOS (eNOS) and neuronal NOS (nNOS) ( P 0.05 ) .Conclusions Is-chemic postconditioning can protect the brain tissue of diabetic rats by inhibiting NOS activity especially iNOS .
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ObjectiveTo investigate the expression and effects of nitric oxide synthase in the Abeta transgenic drosophila.MethodsmRNA was obtained from 20 Abeta transgenic drosophila and 20 wild type drosophila,and cDNA was synthesized by reverse transcription.The expression of nitric oxide synthase mRNA was detected by RT-PCR.ResultsThe expression of nitric oxide synthase mRNA (0.020 ± 0.006)in the Abeta transgenic drosophila was significantly lower than that in the wild type drosophila ( 1.000 ±0.149),there had significant difference between Abeta transgenic drosophila and wild type drosophila ( P ＜ 0.05)..ConclusionsNitric oxide generated by nitric oxide synthase was an important messenger molecule,and it was closely related to Alzheimer's disease.