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Volume 13, Issue 3 (12-2023)
cmja 2023, 13(3): 11-20 |
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Eskandari N, Azarbayjani M A, Matinhomaee H, Fatolahi H. The Effect of Date Pollen Extract/Testosterone and Resistance Training on Gene Expression and Synthesis Rate of Occludin and Cadherin Proteins of Blood-Spinal Cord Barrier in Male Rats. cmja 2023; 13 (3) :11-20
URL: http://cmja.arakmu.ac.ir/article-1-946-en.html
URL: http://cmja.arakmu.ac.ir/article-1-946-en.html
1- Department of Exercise Physiology, Central Tehran Branch, Islamic Azad University, Tehran, Iran
2- Department of Exercise Physiology, Central Tehran Branch, Islamic Azad University, Tehran, Iran , m_azarbayjani@iauctb.ac.ir
3- Department of Physical Education, Pardis Branch, Islamic Azad University, Pardis, Iran
2- Department of Exercise Physiology, Central Tehran Branch, Islamic Azad University, Tehran, Iran , m_azarbayjani@iauctb.ac.ir
3- Department of Physical Education, Pardis Branch, Islamic Azad University, Pardis, Iran
Keywords: Blood-spinal barrier [MeSH], Tight junction protein [MeSH], Adherens junction protein [MeSH], Resistance training [MeSH], Date pollen extract [MeSH], testosterone [MeSH]
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Table 1. The results of the two-way analysis of variance test in relation to gene expression and protein expression of occludin
Table 2. The results of the two-way analysis of variance test in relation to gene expression and protein expression of cadherin
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INTRODUCTION
The brain-blood barrier (BBB) and spinal cord-blood barrier (BSCB) play an important role in protecting the central nervous system against external factors such as hormones, toxins, and pathogens. The regular functioning of both barriers is regulated by a combination of protein structures called tight junctions (TJs) and adherens junctions (AJs). The most important of these proteins include occludin and claudin 1, 2, 5, 8, 13 and binding adhesive molecules. Apparently, a wide range of signaling processes including PKC and GTPases and cytokine activation are involved in the regulation of occludin and cadherin. Changes in gene expression and protein expression of the spinal-blood barrier play an important role in the transmission and development of neuro-inflammatory responses. In order to increase the knowledge related to the role of environmental interventions affecting the tight junction and adhesive proteins of this barrier, this study investigated the effect of four weeks of resistance training and date pollen/testosterone extract on gene expression and protein expression of occludin and cadherin in male rats.
METHODS
In an experimental design, 30 male Wistar rats randomly divided into 6 groups: Control; Resistance training; DPP extract; Testosterone; DPP extract+ resistance training; Testosterone+ resistance training. The amount of 100 mg/kg date pollen extract was fed to mice by gavage for 4 weeks. Also, synthetic testosterone with a dose of 2 mg/kg was used subcutaneously as a positive control. Resistance training consisted of a four-week increasing training period on a 50-step ladder with 8 to 12 repetitions per session. After the intervention, the expression level of occludin and cadhin genes in spinal cord tissue was evaluated using Real-time PCR method according to the steps of preparation, opening of RNAs, reverse transcriptase reaction, PCR reaction and product analysis. Also, the amount of expression of occludin and cadherin proteins was measured using western blot method according to the instructions of Tatsuta et al. in three stages of separation based on the size of proteins, transfer to paper membrane and labeling of target proteins using primary and secondary antibodies. Central tendency and dispersion indices were used to describe the collected data. Also, two-way analysis of variance was used to investigate the main effect of exercise, synthetic testosterone/palm pollen extract and the interactive effect of exercise and drug on dependent variables. The results are reported at a significance level of 0.05 with figures and graphs obtained from Excel and SPSS software.
RESULTS
Histological staining showed an increase in the amount of lipolysis, a decrease in the number and size of inflammatory cells in the nervous tissue of the testosterone and interaction groups. A summary of the effects of resistance training, date pollen/testosterone extract and their interactive effect on gene expression and protein expression of occludin and cadherin in male rats is presented in Tables 1 and 2. The protein synthesis rate of occludin (P=0.016) and cadherin (P=0.012) was increased only in the group receiving synthetic testosterone. However, the highest expression of occludin and cadherin protein occurred when resistance training was combined with testosterone supplementation (P<0.05).
The brain-blood barrier (BBB) and spinal cord-blood barrier (BSCB) play an important role in protecting the central nervous system against external factors such as hormones, toxins, and pathogens. The regular functioning of both barriers is regulated by a combination of protein structures called tight junctions (TJs) and adherens junctions (AJs). The most important of these proteins include occludin and claudin 1, 2, 5, 8, 13 and binding adhesive molecules. Apparently, a wide range of signaling processes including PKC and GTPases and cytokine activation are involved in the regulation of occludin and cadherin. Changes in gene expression and protein expression of the spinal-blood barrier play an important role in the transmission and development of neuro-inflammatory responses. In order to increase the knowledge related to the role of environmental interventions affecting the tight junction and adhesive proteins of this barrier, this study investigated the effect of four weeks of resistance training and date pollen/testosterone extract on gene expression and protein expression of occludin and cadherin in male rats.
METHODS
In an experimental design, 30 male Wistar rats randomly divided into 6 groups: Control; Resistance training; DPP extract; Testosterone; DPP extract+ resistance training; Testosterone+ resistance training. The amount of 100 mg/kg date pollen extract was fed to mice by gavage for 4 weeks. Also, synthetic testosterone with a dose of 2 mg/kg was used subcutaneously as a positive control. Resistance training consisted of a four-week increasing training period on a 50-step ladder with 8 to 12 repetitions per session. After the intervention, the expression level of occludin and cadhin genes in spinal cord tissue was evaluated using Real-time PCR method according to the steps of preparation, opening of RNAs, reverse transcriptase reaction, PCR reaction and product analysis. Also, the amount of expression of occludin and cadherin proteins was measured using western blot method according to the instructions of Tatsuta et al. in three stages of separation based on the size of proteins, transfer to paper membrane and labeling of target proteins using primary and secondary antibodies. Central tendency and dispersion indices were used to describe the collected data. Also, two-way analysis of variance was used to investigate the main effect of exercise, synthetic testosterone/palm pollen extract and the interactive effect of exercise and drug on dependent variables. The results are reported at a significance level of 0.05 with figures and graphs obtained from Excel and SPSS software.
RESULTS
Histological staining showed an increase in the amount of lipolysis, a decrease in the number and size of inflammatory cells in the nervous tissue of the testosterone and interaction groups. A summary of the effects of resistance training, date pollen/testosterone extract and their interactive effect on gene expression and protein expression of occludin and cadherin in male rats is presented in Tables 1 and 2. The protein synthesis rate of occludin (P=0.016) and cadherin (P=0.012) was increased only in the group receiving synthetic testosterone. However, the highest expression of occludin and cadherin protein occurred when resistance training was combined with testosterone supplementation (P<0.05).
Table 1. The results of the two-way analysis of variance test in relation to gene expression and protein expression of occludin
| Factor | Gene Expression | Protein Expression | ||||
| F | sig | Es | F | sig | Es | |
| Resistance training | 10.081 | 0.010 | 0.373 | 0.951 | 0.404 | 0.096 |
| Drug | 13.002 | 0.005 | 0.430 | 5.296 | 0.021 | 0.261 |
| Resistance training* Drug | 13.992 | 0.001 | 0.672 | 8.318 | 0.001 | 0.839 |
Table 2. The results of the two-way analysis of variance test in relation to gene expression and protein expression of cadherin
| Factor | Gene Expression | Protein Expression | ||||
| F | sig | Es | F | sig | Es | |
| Resistance training | 6.408 | 0.015 | 0.265 | 0.783 | 0.647 | 0.079 |
| Drug | 9.700 | 0.004 | 0.441 | 5.088 | 0.033 | 0.252 |
| Resistance training* Drug | 15.082 | 0.001 | 0.728 | 8.624 | 0.001 | 0.862 |
CONCLUSION
The findings of this research showed the amount of reversible physiological change in gene expression and protein expression of occludin and cadherin in BSCB barrier after four weeks of resistance training and consumption of DPP/testosterone extract. Apparently, these interventions have a greater effect on the gene expression of tight junction proteins than adhesive junctions. However, their effects on the expression of occludin and cadherin proteins are completely similar. In spite of the fact that DPP extract can be a suitable alternative for increasing the gene expression of proteins compared to testosterone and resistance training, the separate consumption of testosterone is superior in terms of increasing the expression of occludin and cadherin proteins. In order to increase the synergistic effect, it is more important to combine resistance training with testosterone.
Compliance with Ethical Guidelines
This article is taken from a research project with ethics code IR.UT.SPORT.REC.1397.028 which has been registered in the University Islamic Azad University, Central Tehran Branch.
Funding
The paper was extracted from the PhD. dissertation of the first authorat the Department of Exercise Physiology, Central Tehran Branch, Islamic Azad University, Tehran, Iran.
Authors' Contributions
All authors have participated in the design, implementation and writing of sections of the present study.
Conflicts of Interest
The authors declared no conflict of interest.
Acknowledgements
The authors would like to thank all the contributors who contributed to this research.
The findings of this research showed the amount of reversible physiological change in gene expression and protein expression of occludin and cadherin in BSCB barrier after four weeks of resistance training and consumption of DPP/testosterone extract. Apparently, these interventions have a greater effect on the gene expression of tight junction proteins than adhesive junctions. However, their effects on the expression of occludin and cadherin proteins are completely similar. In spite of the fact that DPP extract can be a suitable alternative for increasing the gene expression of proteins compared to testosterone and resistance training, the separate consumption of testosterone is superior in terms of increasing the expression of occludin and cadherin proteins. In order to increase the synergistic effect, it is more important to combine resistance training with testosterone.
Compliance with Ethical Guidelines
This article is taken from a research project with ethics code IR.UT.SPORT.REC.1397.028 which has been registered in the University Islamic Azad University, Central Tehran Branch.
Funding
The paper was extracted from the PhD. dissertation of the first authorat the Department of Exercise Physiology, Central Tehran Branch, Islamic Azad University, Tehran, Iran.
Authors' Contributions
All authors have participated in the design, implementation and writing of sections of the present study.
Conflicts of Interest
The authors declared no conflict of interest.
Acknowledgements
The authors would like to thank all the contributors who contributed to this research.
Type of Study: Research |
Subject:
Physiology
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