scholarly journals Resveratrol reduced the detrimental effects of malondialdehyde on human endothelial cells

2021 ◽  
Vol 13 (2) ◽  
pp. 131-140
Author(s):  
Mehdi Hassanpour ◽  
Çıgır Biray Avci ◽  
Reza Rahbarghazi ◽  
Aysa Rezabakhsh ◽  
Alireza Nourazarian ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Endothelial Cells ◽  
Surface Plasmon Resonance ◽  
Chromatin Remodeling ◽  
Tyrosine Kinases ◽  
Receptor Tyrosine Kinases ◽  
Pcr Array ◽  
Human Endothelial Cells ◽  
Chain Reaction ◽  
Polymerase Chain

Introduction: According to the statistics, vascular injury occurs during the onset of diabetic changes after the production of several byproducts. Many authorities have focused to find an alternative therapy for diabetic patients. In this study, we investigated the therapeutic effects of natural polyphenol like resveratrol on human endothelial cells exposed to malondialdehyde for 48 hours. Methods: Human Umbilical Vein Endothelial Cells were randomly classified into four groups;control, malondialdehyde (2.5 mM), resveratrol (100 μM), and cells received the combined regime for 48 hours. Cell viability was determined by 3-(4, 5-dimethyl thiazol-2-yl) 2, 5-diphenyl-tetrazoliumbromide (MTT) assay. Griess reaction was performed to measure the content of Nitric oxide (NO).Apoptosis was studied by using real-time polymerase chain reaction (RT-PCR) and western blotting assays. Levels of receptor tyrosine kinases like VEGFR-1, -2, Tie-1, and -2 were analyzed by enzyme-linked immunosorbent assay(ELISA). The affinity of resveratrol and malondialdehyde to serum albumin was measured by Surface Plasmon Resonance Assay. Any changes in chromatin remodeling were detected by PCR array analysis. Results: Resveratrol reduced cytotoxicity and NO content inside cells induced by malondialdehyde(MDA) (P < 0.05). Endothelial cell apoptosis was decreased by the reduction of pro-apoptotic factor Bax and increase of Bcl-2 following the incubation with resveratrol (P < 0.05). MDA-induced receptor tyrosine kinases increase was inhibited by resveratrol and reached near-to-normal levels (P < 0.05).Surface Plasmon Resonance revealed a higher affinity of resveratrol to albumin compared to the malondialdehyde-albumin complex. Polymerase chain reaction (PCR) array revealed the potency of resveratrol in chromatin remodeling following the treatment with malondialdehyde (P < 0.05). Conclusion: Based on our findings, resveratrol has the potential to decrease diabetic vascular injury induced by lipid byproducts such as MDA.

Expression of Endothelin Peptides and Mrna in the Human Heart

1996 ◽  
Vol 90 (1) ◽  
pp. 37-46 ◽  
Author(s):  
C. Plumpton ◽  
M. J. Ashby ◽  
R. E. Kuc ◽  
G. O'Reilly ◽  
A. P. Davenport
Keyword(s):  
Polymerase Chain Reaction ◽  
Endothelial Cells ◽  
Reverse Transcriptase ◽  
Human Heart ◽  
Immunocytochemical Staining ◽  
Chain Reaction ◽  
Endothelin 1 ◽  
Terminal Fragment ◽  
Polymerase Chain ◽  
Endothelin 3

1. We have examined the expression of endothelin isoforms and their precursors in the human heart using RIA, HPLC, immunocytochemistry and reverse transcriptase—polymerase chain reaction assays. 2. Highly specific RIAs were used to measure the levels of mature endothelin and big endothelin-1 immunoreactivity in extracts of human right ventricle. There was no significant difference between samples from patients with ischaemic heart disease and idiopathic dilated cardiomyopathy. 3. HPLC coupled with RIAs allowed the separation and identification of the three mature isoforms of endothelin, big endothelin-1 and the C-terminal fragment of big endothelin-1. In extracts of human endocardial endothelial cells, peaks of immunoreactivity that co-eluted with authentic endothelin-1, big endothelin-1 and C-terminal fragment were found. 4. Intense immunocytochemical staining of mature endothelin immunoreactivity was detected in the cytoplasm of endothelial cells of all regions of the heart tested. Big endothelin-1 immunoreactivity mirrored that of the mature peptide and, in two of three individuals tested, big endothelin-2 immunoreactivity was also detected. No big endothelin-3 immunoreactivity was detected in any of the tissues examined. 5. Reverse transcriptase—polymerase chain reaction assays demonstrated endothelin-1 and endothelin-2 mRNA in all three samples of human left ventricle tested. In two of the individuals, additional bands were also detected with the endothelin-2 primers which corresponded to splice variants. There was no evidence for the expression of endothelin-3 mRNA. 6. These data suggest that endothelin-1 is the predominant isoform of endothelin in the human heart and is probably largely synthesized by the endothelial cells within the heart. If released from the endothelial cells in vivo, this potent cardiotonic peptide may play an important paracrine role in human cardiovascular function.

scholarly journals Rapid Simultaneous Detection of Multiple Micro-Ribonucleic Acids By Surface Plasmon Resonance Array in Cell Lysates of Myelodysplastic Syndromes Patients

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 5605-5605
Author(s):  
Jan E. Dyr ◽  
Jaroslav Cermak ◽  
Hana Vaisocherova ◽  
Zdenek Krejcik ◽  
Hana Sipova ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Surface Plasmon Resonance ◽  
Real Time ◽  
Surface Plasmon ◽  
Myelodysplastic Syndromes ◽  
Simultaneous Detection ◽  
High Capacity ◽  
Chain Reaction ◽  
Mirna Detection ◽  
Polymerase Chain

Abstract Introduction MicroRNAs (miRNAs) are involved in the control of hematopoiesis, their deregulation also appears to play role in the pathogenesis of several hematopoietic diseases. Multiple miRNAs have been reported to be abnormally expressed in hematologic cancers; moreover, specific miRNA expression profiles have been proposed as diagnostic and prognostic markers in various hematologic malignancies, including Myelodysplastic Syndromes (MDS). MDS are a heterogeneous family of clonal disorders of hematopoietic stem cells. They are characterized by ineffective hematopoiesis and frequent leukemic progression. It has been shown that the mature erythrocytes are rich in diverse miRNAs species, even though they lack ribosomal and other large-size RNAs. miRNAs are expressed during erythrocyte differentiation and have an important role in erythropoiesis and mRNA degradation. Methods Four miRNAs, i.e. miR-16, miR-181, miR-34a, and miR-125b were selected as a model set of potential MDS biomarkers and evaluated using a surface plasmon resonance imaging (SPRi) biosensor and real-time PCR. The sensor calibration curves were measured with miRNAs spiked in a complex erythrocyte lysate. The total RNA was extracted from erythrocyte lysate using the acid guanidinium-thiocyanate-phenol-chloroform method. The levels of selected miRNAs were measured in all samples by quantitative reverse transcription - real-time polymerase chain reaction. Results A high-capacity array SPRi system for rapid simultaneous detection of multiple miRNAs in erythrocyte lysate was developed and demonstrated at the Institute of Photonics and Electronics. The ultra-low fouling functionalizable poly(carboxybetaine acrylamide) (pCBAA) brushes-coated gold surfaces of the SPR sensor were shown to reduce the non-specific interaction between surface of the sensor and sample. A two-step miRNA detection assay for multiplexed miRNA detection in erythrocyte lysate was demonstrated in which hybridization of probe-functionalized pCBAA with target miRNA bound to biotinylated oligonucleotide probes was followed with capture of streptavidin-functionalized gold nanoparticles to biotinylated probes. In preliminary experiments, the array was shown to be capable of detecting multiple miRNAs spiked in erythrocyte lysate without the need for complex lysate sample pretreatment at concentrations as low as 0.5 pM in less than 45 minutes. Conclusions Using the newly developed high-capacity array SPRi system we have found significantly increased levels of miR-16 in erythrocyte lysate samples (at the concentration range of ~10-100 pM) compared to other miRNAs tested within this study. The results were confirmed by a reference real-time polymerase chain reaction method. In addition, our results indicate an over-expression of miR-16 in erythrocyte lysate samples of MDS patients with both Refractory Cytopenia with Multilineage Dysplasia (RCMD)and Refractory Anemia with Excess Blasts (RAEB) diagnosis. This clearly demonstrates the potential of SPRi array technology for clinical applications. Disclosures No relevant conflicts of interest to declare.

A Continuous Flow PCR Array Microfluidic Chip Applied for Simultaneous Amplification of Target Genes of Periodontal Pathogens

Lab on a Chip ◽  
2022 ◽  
Author(s):  
Bo Yang ◽  
Ping Wang ◽  
Zhenqing Li ◽  
Chunxian Tao ◽  
Qingxiang You ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Microfluidic Chip ◽  
Continuous Flow ◽  
Target Genes ◽  
Pcr Array ◽  
Periodontal Pathogens ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Time Required

The concept of time to place conversion makes continuous flow polymerase chain reaction (CF-PCR) microfluidic chip an ideal way to reduce the time required for amplification of target genes, however,...

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