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Effect of acute hypoxia on CXCR4 gene expression in C57BL/6 mouse bone marrow-derived mesenchymal stem cells.

Kadivar M, Alijani N, Farahmandfar M, Rahmati S, Ghahhari NM, Mahdian R - Adv Biomed Res (2014)

Bottom Line: Moreover, the relative gene expression of CXCR4 was decreased after hypoxia-reoxygenation by more than 80% in 4 h (0.136 ± 0.018) and 24 h (12.77 ± 0.707) groups.The results suggest that CXCR4 expression in MSCs decreases upon acute hypoxic stress.This difference could have resulted from the cells being compatible with low oxygen metabolism.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Pasteur Institute of Iran, Tehran, Iran.

ABSTRACT

Background: One of the most important stimuli in stem cell biology is oxygen. Chemokine receptor 4 (CXCR4) plays a crucial role in the migration and homing of stem cells. In this study, mesenchymal stem cells (MSCs) were exposed to 1% oxygen to investigate the effect of acute hypoxia on CXCR4 gene expression.

Materials and methods: MSCs were isolated from C57BL/6 mouse bone marrow and were identified and expanded in normoxic culture. Cells were incubated at 37°C under 1% hypoxic conditions for periods of 4, 8, 16, 24, and 48 h. After hypoxia preconditioning, the cells were placed in normoxic condition for 8 h to achieve cellular hypoxia-reoxygenation. To assess the level of CXCR4 gene expression, real-time quantitative reverse transcription-polymerase chain reaction was carried out for each group.

Results: Data from statistical analysis illustrated that exposure of MSCs to acute hypoxic condition down-regulates CXCR4 expression with the maximum under-expression observed in 4 h (0.91 ± 0.107) and 8 h (50 ± 2.98) groups. Moreover, the relative gene expression of CXCR4 was decreased after hypoxia-reoxygenation by more than 80% in 4 h (0.136 ± 0.018) and 24 h (12.77 ± 0.707) groups.

Conclusion: The results suggest that CXCR4 expression in MSCs decreases upon acute hypoxic stress. Furthermore, hypoxia-reoxygenated MSCs showed decreased expression of CXCR4, compared to cells subjected to acute hypoxia. This difference could have resulted from the cells being compatible with low oxygen metabolism. In summary, before the therapeutic application of MSCs, it should be regarded as a necessity to optimize the oxygen concentration in these cells, as it is a critical factor in modulating CXCR4 expression.

No MeSH data available.


Related in: MedlinePlus

Real-time reverse transcription-polymerase chain reaction analysis of the mRNA expression of CXCR4 receptor gene: (a) comparison between hypoxia group and normoxia group; (b) comparison between hypoxia-reoxygenation group and normoxia group; (c) comparison between hypoxia group and hypoxia-reoxygenation groups. Control groups were treated under normoxia condition. Data show relative mRNA expression of CXCR4 normalized against 18S rRNA. Values are mean ± standard error of mean ***P < 0.001
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Figure 4: Real-time reverse transcription-polymerase chain reaction analysis of the mRNA expression of CXCR4 receptor gene: (a) comparison between hypoxia group and normoxia group; (b) comparison between hypoxia-reoxygenation group and normoxia group; (c) comparison between hypoxia group and hypoxia-reoxygenation groups. Control groups were treated under normoxia condition. Data show relative mRNA expression of CXCR4 normalized against 18S rRNA. Values are mean ± standard error of mean ***P < 0.001

Mentions: As shown in Figure 4a, after exposure of cells to short-term hypoxia (1% O2) for 4, 8, 16, 24, and 48 h, CXCR4 relative gene expression was decreased in comparison to normoxic cells as control samples. The maximum decrease was observed in 4 h (0.91 ± 0.107) group, while the minimum decrease was in 8 h (50 ± 2.98) group. Figure 4b shows that hypoxia-reoxygenation treatment leads to CXCR4 gene under-expression, with the maximum decrease measured in 4 h (0.136 ± 0.018) group and the minimum decrease in 24 h (12.77 ± 0.707) group. Figure 4c presents a comparison between different hypoxia groups and their related hypoxia-reoxygenation groups. The differences between hypoxia and normoxia groups, hypoxia-reoxygenation and normoxia groups were significant as calculated by One-Way Anova. Also differences between hypoxia and hypoxia-reoxygenation groups were significant, as calculated by Student's t-test (P < 0.001).


Effect of acute hypoxia on CXCR4 gene expression in C57BL/6 mouse bone marrow-derived mesenchymal stem cells.

Kadivar M, Alijani N, Farahmandfar M, Rahmati S, Ghahhari NM, Mahdian R - Adv Biomed Res (2014)

Real-time reverse transcription-polymerase chain reaction analysis of the mRNA expression of CXCR4 receptor gene: (a) comparison between hypoxia group and normoxia group; (b) comparison between hypoxia-reoxygenation group and normoxia group; (c) comparison between hypoxia group and hypoxia-reoxygenation groups. Control groups were treated under normoxia condition. Data show relative mRNA expression of CXCR4 normalized against 18S rRNA. Values are mean ± standard error of mean ***P < 0.001
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4260281&req=5

Figure 4: Real-time reverse transcription-polymerase chain reaction analysis of the mRNA expression of CXCR4 receptor gene: (a) comparison between hypoxia group and normoxia group; (b) comparison between hypoxia-reoxygenation group and normoxia group; (c) comparison between hypoxia group and hypoxia-reoxygenation groups. Control groups were treated under normoxia condition. Data show relative mRNA expression of CXCR4 normalized against 18S rRNA. Values are mean ± standard error of mean ***P < 0.001
Mentions: As shown in Figure 4a, after exposure of cells to short-term hypoxia (1% O2) for 4, 8, 16, 24, and 48 h, CXCR4 relative gene expression was decreased in comparison to normoxic cells as control samples. The maximum decrease was observed in 4 h (0.91 ± 0.107) group, while the minimum decrease was in 8 h (50 ± 2.98) group. Figure 4b shows that hypoxia-reoxygenation treatment leads to CXCR4 gene under-expression, with the maximum decrease measured in 4 h (0.136 ± 0.018) group and the minimum decrease in 24 h (12.77 ± 0.707) group. Figure 4c presents a comparison between different hypoxia groups and their related hypoxia-reoxygenation groups. The differences between hypoxia and normoxia groups, hypoxia-reoxygenation and normoxia groups were significant as calculated by One-Way Anova. Also differences between hypoxia and hypoxia-reoxygenation groups were significant, as calculated by Student's t-test (P < 0.001).

Bottom Line: Moreover, the relative gene expression of CXCR4 was decreased after hypoxia-reoxygenation by more than 80% in 4 h (0.136 ± 0.018) and 24 h (12.77 ± 0.707) groups.The results suggest that CXCR4 expression in MSCs decreases upon acute hypoxic stress.This difference could have resulted from the cells being compatible with low oxygen metabolism.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Pasteur Institute of Iran, Tehran, Iran.

ABSTRACT

Background: One of the most important stimuli in stem cell biology is oxygen. Chemokine receptor 4 (CXCR4) plays a crucial role in the migration and homing of stem cells. In this study, mesenchymal stem cells (MSCs) were exposed to 1% oxygen to investigate the effect of acute hypoxia on CXCR4 gene expression.

Materials and methods: MSCs were isolated from C57BL/6 mouse bone marrow and were identified and expanded in normoxic culture. Cells were incubated at 37°C under 1% hypoxic conditions for periods of 4, 8, 16, 24, and 48 h. After hypoxia preconditioning, the cells were placed in normoxic condition for 8 h to achieve cellular hypoxia-reoxygenation. To assess the level of CXCR4 gene expression, real-time quantitative reverse transcription-polymerase chain reaction was carried out for each group.

Results: Data from statistical analysis illustrated that exposure of MSCs to acute hypoxic condition down-regulates CXCR4 expression with the maximum under-expression observed in 4 h (0.91 ± 0.107) and 8 h (50 ± 2.98) groups. Moreover, the relative gene expression of CXCR4 was decreased after hypoxia-reoxygenation by more than 80% in 4 h (0.136 ± 0.018) and 24 h (12.77 ± 0.707) groups.

Conclusion: The results suggest that CXCR4 expression in MSCs decreases upon acute hypoxic stress. Furthermore, hypoxia-reoxygenated MSCs showed decreased expression of CXCR4, compared to cells subjected to acute hypoxia. This difference could have resulted from the cells being compatible with low oxygen metabolism. In summary, before the therapeutic application of MSCs, it should be regarded as a necessity to optimize the oxygen concentration in these cells, as it is a critical factor in modulating CXCR4 expression.

No MeSH data available.


Related in: MedlinePlus