Abstract
Anaemia occurs frequently in patients with heart failure and its current treatment lacks clear targets. Emerging evidence suggests that erythroid progenitor cell expansion is an integral part of physiological response to anaemia associated with chronic stress. Understanding the underlying mechanism may provide a novel approach to anaemia management. In this study, we aimed to examine a role for nitric oxide (NO) in the regulation of bone marrow erythroid progenitor response to chronic stress. For this purpose, adult male mice were subjected to 2 h daily restraint stress for 7 or 14 consecutive days. The role of NO was assessed by subcutaneous injection with NG-nitro-l-arginine methyl ester, 30 min prior to each restraint. Chronic exposure to stress resulted in significantly increased number of bone marrow erythroid progenitors, and blockade of NO biosynthesis prior to daily stress completely prevented stress-induced erythroid progenitor cell expansion. Furthermore, chronic stress exposure led to altered expression of neural, endothelial and inducible nitric oxide synthases (NOS) in the bone marrow, both on mRNA and protein level. Decreased expression of neural and endothelial NOS, as well as reduced expression of NF-kappaB/p65 in bone marrow nuclear cell fraction, was accompanied by elevated bone marrow expression of inducible NOS in chronically stressed animals. This is the first study to demonstrate a role for NO in adaptive response of erythroid progenitors to chronic stress. Targeting NO production may be beneficial to improve bone marrow dysfunction and reduced erythroid progenitor cell expansion in chronic heart failure patients.
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Abbreviations
- NO:
-
Nitric oxide
- nNOS:
-
Neuronal nitric oxide synthase
- iNOS:
-
Inducible nitric oxide synthase
- eNOS:
-
Endothelial nitric oxide synthase
- NF-kappaB:
-
Nuclear factor kappa B
- L-NAME:
-
Nω-nitro-l-arginine-methyl ester
- BFU-E:
-
Burst-forming unit erythroid cells
- CFU-E:
-
Colony-forming unit erythroid cells
- REST:
-
Relative expression Software Tool
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The authors would like to thank Mrs Snežana Marković for technical support.
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This work was supported by the Grants (175053, 175062) from the Ministry of Education, Science and Technological Development of Republic of Serbia.
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GJ, MB and SVP conceived the study; SVP, MB, MM and SM conducted in vivo experiments; DM performed Western blot analysis; OMA performed immunohistochemical analysis; SVP and SM performed qRT-PCR analysis; SVP, MB, DM, OMA, VČ and MM analysed data; SVP and MB wrote the manuscript; GJ and VČ contributed to review and revision of final manuscript. All authors approved the final version of the manuscript.
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The experimental protocol was approved by the Ethic Committee of the Institute for Medical Research, University of Belgrade, Serbia (No O112-1/12), according to the National Law on Animal Welfare that is consistent with guidelines for animal research and principles of the European Convention for the Protection of Vertebrate Animals Used for Experimental and Other Purposes (Official Daily N. L 358/1–358/6, 18, December 1986) and Directive on the protection of animals used for scientific purposes (Directive 2010/63/EU of the European Parliament and of the Council, 22 September 2010).
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418_2020_1856_MOESM1_ESM.tif
Supplementary file1 The effects of chronic stress on BFU-E and CFU-E in the bone marrow. Data are expressed as mean ± SEM (n = 8/group). **p < 0.01; *p <0.05. C-control mice, R7 and R14 - animals stressed for 7 or 14 days (TIF 265 kb)
418_2020_1856_MOESM2_ESM.tif
Supplementary file2 The effects of chronic restraint stress and L-NAME on blood haemoglobin levels. C – controls; R – mice subjected to daily restraint stress for 7 consecutive days; L-NAME+R – animals treated with L-NAME prior to daily restraint; L-NAME – mice received only the daily dose of L-NAME. Data are expressed as mean ± SEM **p <0.01 (TIF 94 kb)
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Vignjević Petrinović, S., Budeč, M., Marković, D. et al. Nitric oxide-dependent expansion of erythroid progenitors in a murine model of chronic psychological stress. Histochem Cell Biol 153, 457–468 (2020). https://doi.org/10.1007/s00418-020-01856-y
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DOI: https://doi.org/10.1007/s00418-020-01856-y