Objective: Atherosclerosis (AS) is one of the most common causes of human death and disability. This study is designed to investigate the roles of aldosterone (Aldo) and oxidized low-density lipoprotein (Ox-LDL) in this disease by clinical data and cell model. Materials and Methods: In this experimental study, clinical data were collected to investigate the Aldo role for the patients with primary aldosteronism or adrenal tumors. Cell viability assay, fluorescence-activated cell sorting (FACS) assay, apoptosis assay, cell aging analysis, and matrigel tube formation assay were performed to detect effects on human umbilical vein endothelial cells (HUVECs) treated with Aldo and/or Ox-LDL. Quantitative polymerase chain reaction (qPCR) and Western blot analysis were performed to figure out critical genes in the process of endothelial cells Dysfunction induced by Aldo and/or Ox-LDL. Results: We found that the Aldo level had a positive correlation with the TG/HDL-C ratio. Endothelial cell growth, angiogenesis, senescence, and apoptosis were significantly affected, and eNOS/Sirt1, the value of Bcl-2/Bax and Angiopoietin1/2 were significantly affected when cells were co-treated by Aldo and Ox-LDL. Conclusion: Elevated Aldo with high Ox-LDL together may accelerate the Dysfunction of HUVEC, and the Ox-LDL, especially for those patients with high Aldo should be well controlled. The assessment of the role of Aldo may provide a theoretical basis for the effective prevention and investigation of a new treatment of AS.

Objective: Patients with diabetes mellitus frequently have chronic wounds or diabetic ulcers as a result of impaired wound healing, which may lead to limb amputation. Human umbilical vein endothelial cell (HUVEC) Dysfunction also delays wound healing. Here, we investigated the mechanism of miR-200b in HUVECs under high glucose conditions and the potential of miR-200b as a therapeutic target. Materials and Methods: In this experimental study, HUVECs were cultured with 5 or 30 mM glucose for 48 hours. Cell proliferation was evaluated by CCK-8 assays. Cell mobility was tested by wound healing and Transwell assays. Angiogenesis was analyzed in vitro Matrigel tube formation assays. Luciferase reporter assays were used to test the binding of miR-200b with Notch1. Results: miR-200b expression was induced by high glucose treatment of HUVECs (P<0. 01), and it significantly repressed cell proliferation, migration, and tube formation (P<0. 05). Notch1 was directly targeted and repressed by miR-200b at both the mRNA and protein levels. Inhibition of miR-200b restored Notch1 expression (P<0. 05) and reactivated the Notch pathway. The effects of miR-200b inhibition in HUVECs could be reversed by treatment with a Notch pathway inhibitor (P<0. 05), indicating that the miR-200b/Notch axis modulates the proliferation, migration, and tube formation ability of HUVECs. Conclusion: Inhibition of miR-200b activated the angiogenic ability of endothelial cells and promoted wound healing through reactivation of the Notch pathway in vitro. miR-200b could be a promising therapeutic target for treating HUVEC Dysfunction.

Background and purpose: Several adverse effects were reported for oral sildenafil. In this research after formulation of sildenafil topical gel, the effect of gel was compared with sildenafil tablet in a double-blind placebo-controled clinical trial. Materials and methods: After choosing the solvent system, several formulations were prepared and the most suitable gel was chosen for clinical trial. The study was a controlled randomized (block-random sampling), double blind, prospective, placebo-controlled trial. 94 patients, with clinically diagnosed erectile Dysfunction were recruited. This includes men with ED of organic, psychogenic, and mixed causes. Patients were divided into two categories of age under and over 50. Six blocks were considered for including patients based on the nature of ED and age. All of the patients with ED from July 2003 to May 2004 diagnosed by one urologist were included in the study. The cases received 1% topical gel of sildenafil and placebo tablet, and the control group received sildenafil tablet (100 mg) and gel base (without drug) as placebo. The tablets were taken one hour before sexual action and the topical gel was used in 0.5g (approximately) on the glans of penis and were massaged for 5min, before the sexual activity. Results: In the case group that topical gel of sildenafil was administered, 5 patients (12.5%) had complete erection, 5 patients had moderate erection and erection was not observed in 30 (75%) of them. In control group, that sildenafil tablet was administered, these results were 28 (70%), 6 (15%) and 6 (15%) respectively. The onset of erection in case group (in-patients with complete erection) was 7.4 ± 3.6 min, but this time was 37.8 ± 14.9 min in control group. Four cases of mild headache were observed in-patients who administered topical gel of sildenafil. This was pain treated before 4 min. Two cases of severe headache were observed in-patients who were administered sildenafil tablet. The disorder on visual function was observed in one patient in control group. One case of severe dyspepsia was observed in this group too. Conclusion: Sildenafil delivery using transdermal formulation can be enhanced by several synthetic or natural percutaneous absorption enhancers, and appears to be a promising approach for the treatment of erectile Dysfunction.  

Objective: Envenomation by heamotoxic snakes constituted a critical health occurrence in the world. Bleeding is the most sever consequence following snake bite with viperid and crothalid snakes. It is believed that the degradation of vascular membrane caused hemorrhage; in contrast, some suggested that direct cytotoxicity has role in endothelial cell disturbances. This study was carried out to evaluate the direct toxicity effect of V. lebetina crude venom on Human Umbilical Vein Endothelial Cells (HUVECs). Methods: The effect of V. lebetina snake venom on HUVECs growth inhibition was determined by MTT assay and neutral red uptake assay. The integrity of cell membrane through LDH release was measured with the Cytotoxicity Detection Kit. Morphological changes of endothelial cells were also evaluated using a phase contrast microscope. Result: In MTT assay, crude venom showed a cytotoxic effect on endothelial cells which was confirmed by the effect observed with neutral red assay. Also, crude venom caused changes in the integrity of cell membrane by LDH release. The morphological alterations enhanced in high concentration results in total cells number reduced. Conclusion: V. lebetina venom showed potential direct cytotoxic effects on human endothelial cells in a manner of concentration-dependent inhibition.

Background & aim: Angiogenesis is the production of new blood vessels from existing vessels. Angiogenesis depends on the balance between its activating and inhibitory factors. Endostatin, a 20-kDa fragment of collagen XVIII, is a member of the angiogenesis inhibitory protein group that inhibits endothelial cell proliferation and migration and tubular-like structure. The use of colorimetric methods by fluorescent probes as a tracer is widespread, so the aim of this study was to determine and detect the binding of anti-angiogenic peptide to human cord endothelial cells using a fluorescence marker. Methods: In the present experimental study, the 27 amino acid fragment of the endostatin protein amino acid domain was labeled with FITC as a fluorescence marker. Gel filtration chromatography with Sephadex G10 was used to separate the bound peptides and finally FTIR method was used to confirm the binding of FITC to the peptide. Human umbilical cord endothelial cells were treated with marker-bound peptides after culturing and studied by fluorescence microscopy to observe peptide binding to their receptor on the surface of these cells. Results: findings by Fluorescence microscopy confirmed binding antiangiogenic peptide to its specific receptor. Various conditions such as concentration, time, and temperature were tested to achieve the proper conditions for marking peptides. Unlabeled FITCs separated by gel filtration Chromatography method with sephedex G10. Human umbilical endothelial cells (HUVECs) are harvested and treated with conjugate samples (FITC-peptide) from gel filtration chromatography. After the required time, the cells were fixed and the binding of labeled peptides to their receptors on HUVECs was analyzed by Fluorescence microscope. Conclusion: The results of fluorescence microscopy confirmed the binding of this antiangiogenic peptide to its specific receptor on endothelial cells. Different experimental conditions such as peptide concentration and FITC and incubation time and temperature were studied to obtain the appropriate conditions for peptide labeling. Free FITCs were separated by conjugated FITC-peptide samples by gel filtration chromatography. Human umbilical cord endothelial cells (HUVEC) cultured with FITC-peptide conjugated samples were treated. The cells were then fixed and examined by fluorescence microscopy.

Background & Aims: Curcumin and thymoquinone are common herbal medicines with low side effects and potent antioxidant, anti-diabetic, and anti-cancer activity. This investigation aimed to evaluate toxicity of curcumin and thymoquinone on HUVECS cell lines via MTT and flow cytometry assays. Materials & Methods: HUVEC cells were cultured in RPMI-1640 medium under standard culture conditions (5% CO2 and 95% humidified air at 37° C). Curcumin and thymoquinone were used at concentrations of 5, 10, 20, 40, 80, 160, 320 μ M and incubated for 24 and 48 hours, then cell viability was assessed using MTT assay. Cell apoptosis was measured by flow cytometry via Annexin V-FITC and propidium iodide kit. Data were analyzed using two-way ANOVA and p <0. 05 was considered significant. Results: The results of this study showed that IC50 for curcumin and thymoquinone was 35μ M and 105 μ M at 24 h, and 32 μ M and 90 μ M at 48 h, respectively (p>0. 999). Apoptosis rates for curcumin and thymoquinone were 40% and 30%, respectively. Conclusion: In this study, we observed that cell viability depended on the concentration of curcumin and thymoquinone. Both compounds induced apoptosis and also the cell viability of HUVEC upon treatment with thymoquinone is more than that of curcumin.

Introduction: Over the last decades, considerable levels of electromagnetic fields (EMFs) have been characterized in the living environment. Recent epidemiological studies on occupational and residential exposure to EMF have shown that 50/60 Hz fields, known as extremely low frequencies (ELF), have various biological effects, such as angiogenesis. This study aimed to investigate the effect of environmental 50 Hz magnetic fields at intensities of 3, 6, 15, 46, 110 and 207 mT on human umbilical vein endothelial cells (HUVECs) as a significant component of angiogenesis process.Materials and Methods: In this study, 43 experimental groups were evaluated, including a control group, 6 sham exposure groups, 18 acute, and 18 chronic exposure groups with different exposure intensities and durations of exposure. Proliferation and viability of HUVECs were examined via cell counting and MTT methods, respectively.Results: No significant changes were observed in the proliferation of HUVECs by 50 Hz magnetic field, while the viability of some acute groups was found to increase. These findings confirmed the theory of "biological window" for magnetic fields.Conclusion: According to the results of this study, since the 50 Hz magnetic field can effect on viability of HUVECs and these cells play a key role in angiogenesis, 50 Hz magnetic fields at the mentioned intensities probably could be effective in the improvement of angiogenesis process.