Home Medicine Osteoblast iron genes: real time PCR and microarray hybridization approach under hyperoxia
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Osteoblast iron genes: real time PCR and microarray hybridization approach under hyperoxia

  • Prihartini Widiyanti EMAIL logo , Hartmut Kuehn and Soetjipto Soetjipto
Published/Copyright: June 25, 2021
Journal of Basic and Clinical Physiology and Pharmacology
From the journal Journal of Basic and Clinical Physiology and Pharmacology Volume 32 Issue 4

Abstract

Objectives

Iron is essential for cell growth, differentiation, electron transfer, and oxygen transport. Hyperoxia may increase the turnover of bone matrix components with a net effect of accelerated bone growth. Although hyperoxia was claimed could increase osteoblast activity, but expression level in possible genes which play role in proliferation is still unclear. This research aims to prove the differences of expression level of transferrin receptor gene and iron regulated transporter and other genes of 7F2 under 24 h normoxia, 24 h hyperoxia, and 48 h hyperoxia and the effect of hyperoxia by using osteoblast cell culture 7F2.

Methods

Reverse transcriptase, real time Polymerase Chain Reaction (PCR), and microarray is used to qualitatively detect gene expression. The computer softwares such as National Center for Biotechnology Information (NCBI) data base, Software Affymetrix, DNA Strider program, Genomatix – DiAlign program, Oligo 5.0 program (Software primer design) from Wojciech & Piotr Rychlik, and Genetyx-Mac version 8.0 have been used to analyze the PCR result.

Results

Under 24 h hyperoxia, there were 3,884 copies of transferrin receptor mRNA per 1,000,000 copies of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) mRNA. After 24 h hyperoxia, 8,325 copies of transferrin receptor mRNA per 1,000,000 GAPDH mRNA copies were found showing 2.1-fold up regulation. After 48 h hyperoxia, there was no significant increase at the level of expression of transferrin receptor mRNA, 8,079 mRNA copies per 1,000,000 copies of mRNA were found (2.0-fold up regulation compared with 24 h normoxia).

Conclusions

It can be concluded that hyperoxia might have an effect on upregulating the expression of some osteoblast genes which might have an impact on osteoblast activity.

Keywords: hyperoxia; osteoblast iron-regulated transporter gene; osteoblast transferrin receptor gene
Corresponding author: Prihartini Widiyanti, Biomedical Engineering, Faculty of Science and Technology, Universitas Airlangga, Campus C, Mulyorejo, 60115, Surabaya, Indonesia; and Institute of Tropical Disease, Universitas Airlangga, Surabaya, Indonesia, E-mail:

Funding source: Deutscher Akademischer Austauschdienst

Award Identifier / Grant number: 331 4 00 017

Acknowledgments

The authors deliver gratitude to Deutscher Akademischer Austauschdienst (DAAD) for the sandwich research scholarship to the Charite University of Clinics and Institute of Biochemistry, Charite Universitatsmedizin, University of Humboldt, Berlin, for supporting facilities. The gratitude also for the Institute of Tropical Disease, Universitas Airlangga for all supports.

  1. Research funding: Deutscher Akademischer Austauschdienst (DAAD) Scholarship. All authors have accepted responsibility for the entire content of this manuscript and approved its submission.

  2. Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.

  3. Competing interests: The authors declare that there is no conflict of interest regarding the publication of this article.

  4. Informed consent: Not applicable.

  5. Ethical approval: Not applicable.

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Received: 2020-11-29
Accepted: 2021-02-21
Published Online: 2021-06-25

© 2021 Walter de Gruyter GmbH, Berlin/Boston

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https://doi.org/10.1515/jbcpp-2020-0471
Keywords for this article
hyperoxia; osteoblast iron-regulated transporter gene; osteoblast transferrin receptor gene

Articles in the same Issue

  1. Frontmatter
  2. Original Articles
  3. Cost of illness of diabetes mellitus in Indonesia: a systematic review
  4. Social media health interventions to improve diabetes mellitus patient outcome: a systematic review
  5. Developing pharmacokinetics–pharmacodynamics model of valproic acid syrup based on prediction of population pharmacokinetics parameter and seizure frequency in Indonesian pediatric epilepsy outpatients
  6. Acetylcholinesterase inhibitory activity of extract and fractions from the root of Rauvolfia serpentina(L.) Bth.ex Kurz
  7. Green tea and its active compound epigallocathechin-3-gallate (EGCG) inhibit neuronal apoptosis in a middle cerebral artery occlusion (MCAO) model
  8. The effects of quercetin on nicotine-induced reward effects in mice
  9. Resveratrol ameliorates physical and psychological stress-induced depressive-like behavior
  10. Translation and cross-cultural adaption of an instrument measuring patient’s well-being under treatment for schizophrenia
  11. Quercetin promotes behavioral recovery and biomolecular changes of melanocortin-4 receptor in mice with ischemic stroke
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  13. Inhibition of Ras and STAT3 activity of 4-(tert-butyl)-N-carbamoylbenzamide as antiproliferative agent in HER2-expressing breast cancer cells
  14. Predicting the molecular mechanism of glucosamine in accelerating bone defect repair by stimulating osteogenic proteins
  15. Larvicidal toxicity and parasporal inclusion of native Bacillus thuringiensis BK5.2 against Aedes aegypti
  16. Synthesis, ADMET predictions, molecular docking studies, and in-vitro anticancer activity of some benzoxazines against A549 human lung cancer cells
  17. Thymoquinone and its derivatives against breast cancer with HER2 positive: in silico studies of ADMET, docking and QSPR
  18. Assessment of patient understanding of their conventional cardiac medicines and herbal prepared/derived products: preliminary survey and interviews with selected community-dwelling elderly patients in the Philippines
  19. The development and validation of the health belief model questionnaire for measuring factors affecting adherence in the elderly with hypertension
  20. Analysis of the side effect of QTc interval prolongation in the bedaquiline regimen in drug resistant tuberculosis patients
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  22. Analysis of HMGB-1 level before and after providing atorvastatin standard therapy in coronary artery disease patients with type-2 diabetes mellitus compared to without type-2 diabetes mellitus
  23. Analysis of matrix metalloproteinase-9 levels among acute heart failure patients with ACE inhibitor therapy (Dr. Soetomo Regional General Hospital, Surabaya)
  24. The correlation between self-related adherence, asthma-related quality of life and control of asthma in adult patients
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  28. The association of FKBP5 polymorphism with asthma susceptibility in asthmatic patients
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  31. Attenuation of hyperplasia in lung parenchymal and colonic epithelial cells in DMBA-induced cancer by administering Andrographis paniculata Nees extract using animal model
  32. N-nitrosodiethylamine induces inflammation of liver in mice
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  35. The effect of Camellia sinensis (green tea) with its active compound EGCG on neuronal cell necroptosis in Rattus norvegicus middle cerebral artery occlusion (MCAO) model
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  38. The effect of various high-fat diet on liver histology in the development of NAFLD models in mice
  39. Fabrication and characterization of bovine hydroxyapatite-gelatin-alendronate scaffold cross-linked by glutaraldehyde for bone regeneration
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  42. Correlation of dietary iron intake and serum iron with thyroid stimulating hormone (TSH) and free thyroxine (FT4) levels in adult hyperthyroid patients
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  45. Role of Centella asiatica and ceramide in skin barrier improvement: a double blind clinical trial of Indonesian batik workers
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  50. Cocrystal formation of loratadine-succinic acid and its improved solubility
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  62. Second internal transcribed spacer (ITS-2) as genetic marker for molecular characterization of Sarcoptes scabiei in rabbits from several areas of East Java, Indonesia
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  66. Antibiotic use on acute respiratory tract infection nonpneumonia and nonspecific diarrhea in Primary Health Care Centre in Banjarbaru City, South Kalimantan, Indonesia
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