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Reactivity of rat bone marrow-derived macrophages to neurotransmitter stimulation in the context of collagen II-induced arthritis.

Muschter D, Göttl C, Vogel M, Grifka J, Straub RH, Grässel S - Arthritis Res. Ther. (2015)

Bottom Line: Additionally, we found a strong reduction in proliferation of BMMs at CIA onset and in the chronic phase of CIA.Apoptosis remained unaffected.We suggest an altered reactivity of BMMs to neurotransmitter stimulation caused by CIA and maybe also by aging.

View Article: PubMed Central - PubMed

Affiliation: Experimental Orthopedics, Centre for Medical Biotechnology, Biopark I, University of Regensburg, Josef-Engert-Str. 9, 93053, Regensburg, Germany. dominique.muschter@klinik.uni-regensburg.de.

ABSTRACT

Introduction: Numerous observations indicate that rheumatoid arthritis (RA) has a bone marrow component. In parallel, local synovial changes depend on neuronal components of the peripheral sympathetic nervous system. Here, we wanted to analyze whether collagen II-induced arthritis (CIA) has an impact on number, adhesion, apoptosis, and proliferation of the macrophage subset of bone marrow cells and how alterations in neurotransmitter microenvironment affect these properties.

Methods: Bone marrow-derived macrophages (BMMs) were isolated from Dark Agouti rats at different stages of CIA, and number, adhesion, caspase 3/7 activity, and proliferation were analyzed in the presence of acetylcholine (ACh), noradrenaline (NA), and vasoactive intestinal peptide (VIP).

Results: Opposed to enhanced CD11b(+) (cluster of differentiation 11b-positive) and EMR1(+) (epidermal growth factor-like module-containing mucin-like hormone receptor-like 1-positive) cells, characterizing the macrophage subset, in native bone marrow of rats with acute inflammatory arthritis, we found decreased numbers of CIA macrophages after enrichment and culture in comparison with healthy (control) animals. Adhesion studies revealed significantly reduced attachment to plastic in acute arthritis and collagen type I and fibronectin in chronic arthritis. Additionally, we found a strong reduction in proliferation of BMMs at CIA onset and in the chronic phase of CIA. Apoptosis remained unaffected. Neurotransmitter stimulation profoundly affected proliferation, adhesion, and apoptosis of BMMs from CIA and control rats, depending on disease time point. Cultured BMMs from CIA and control animals expressed neurotransmitter receptors for ACh, VIP and NA, but the expression profile seemed not to be affected by CIA.

Conclusions: Induction of CIA distinctly inhibits proliferation of BMMs in low- and non-inflammatory phases and reduces attachment to plastic at the acute inflammatory arthritis stage and adhesion to collagen I and fibronectin at the chronic stage. Influence of neurotransmitter stimulation on adhesion, apoptosis, and proliferation is altered by CIA depending on disease stage. We suggest an altered reactivity of BMMs to neurotransmitter stimulation caused by CIA and maybe also by aging.

No MeSH data available.


Related in: MedlinePlus

Caspase 3/7 apoptosis assay of pre-cultured BMMs. Pre-cultured BMMs from control and CIA animals which were serum-deprived for 24 h were incubated with caspase 3/7 reagent for 6–10 h. Percentage of apoptosis of CIA BMMs compared with BMMs from controls is presented under (a). Influence of ACh and VIP on apoptosis induction in BMMs from control and CIA rats during the time course of arthritis is shown as percentage to non-stimulated macrophages (non-stimulated = dotted line) (b). The respective results for noradrenergic stimulation are shown under (c). N (control/CIA) = day 10 (9/10), day 15 (12/9), day 20 (10/10), and day 40 (11/12). Neurotransmitter stimulation N (control/CIA) = day 10 (6/6), day 15 (6/3), day 20 (6/6), and day 40 (8/8) for ACh, VIP, and NA (10−6 M, 10−8 M). Data are expressed as mean ± standard error of the mean. *P < 0.05 neurotransmitter stimulation versus non-stimulated macrophages; #P < 0.05 control cells versus CIA cells. ACh acetylcholine, BMM bone marrow-derived macrophage, CIA collagen II-induced arthritis, NA noradrenaline, VIP vasoactive intestinal peptide
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Fig5: Caspase 3/7 apoptosis assay of pre-cultured BMMs. Pre-cultured BMMs from control and CIA animals which were serum-deprived for 24 h were incubated with caspase 3/7 reagent for 6–10 h. Percentage of apoptosis of CIA BMMs compared with BMMs from controls is presented under (a). Influence of ACh and VIP on apoptosis induction in BMMs from control and CIA rats during the time course of arthritis is shown as percentage to non-stimulated macrophages (non-stimulated = dotted line) (b). The respective results for noradrenergic stimulation are shown under (c). N (control/CIA) = day 10 (9/10), day 15 (12/9), day 20 (10/10), and day 40 (11/12). Neurotransmitter stimulation N (control/CIA) = day 10 (6/6), day 15 (6/3), day 20 (6/6), and day 40 (8/8) for ACh, VIP, and NA (10−6 M, 10−8 M). Data are expressed as mean ± standard error of the mean. *P < 0.05 neurotransmitter stimulation versus non-stimulated macrophages; #P < 0.05 control cells versus CIA cells. ACh acetylcholine, BMM bone marrow-derived macrophage, CIA collagen II-induced arthritis, NA noradrenaline, VIP vasoactive intestinal peptide

Mentions: Changes in apoptotic activity could also explain the reduced number of CIA BMMs (Fig. 5). (For more detailed data, see Additional file 4: Table S2E.)Fig. 5


Reactivity of rat bone marrow-derived macrophages to neurotransmitter stimulation in the context of collagen II-induced arthritis.

Muschter D, Göttl C, Vogel M, Grifka J, Straub RH, Grässel S - Arthritis Res. Ther. (2015)

Caspase 3/7 apoptosis assay of pre-cultured BMMs. Pre-cultured BMMs from control and CIA animals which were serum-deprived for 24 h were incubated with caspase 3/7 reagent for 6–10 h. Percentage of apoptosis of CIA BMMs compared with BMMs from controls is presented under (a). Influence of ACh and VIP on apoptosis induction in BMMs from control and CIA rats during the time course of arthritis is shown as percentage to non-stimulated macrophages (non-stimulated = dotted line) (b). The respective results for noradrenergic stimulation are shown under (c). N (control/CIA) = day 10 (9/10), day 15 (12/9), day 20 (10/10), and day 40 (11/12). Neurotransmitter stimulation N (control/CIA) = day 10 (6/6), day 15 (6/3), day 20 (6/6), and day 40 (8/8) for ACh, VIP, and NA (10−6 M, 10−8 M). Data are expressed as mean ± standard error of the mean. *P < 0.05 neurotransmitter stimulation versus non-stimulated macrophages; #P < 0.05 control cells versus CIA cells. ACh acetylcholine, BMM bone marrow-derived macrophage, CIA collagen II-induced arthritis, NA noradrenaline, VIP vasoactive intestinal peptide
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4496866&req=5

Fig5: Caspase 3/7 apoptosis assay of pre-cultured BMMs. Pre-cultured BMMs from control and CIA animals which were serum-deprived for 24 h were incubated with caspase 3/7 reagent for 6–10 h. Percentage of apoptosis of CIA BMMs compared with BMMs from controls is presented under (a). Influence of ACh and VIP on apoptosis induction in BMMs from control and CIA rats during the time course of arthritis is shown as percentage to non-stimulated macrophages (non-stimulated = dotted line) (b). The respective results for noradrenergic stimulation are shown under (c). N (control/CIA) = day 10 (9/10), day 15 (12/9), day 20 (10/10), and day 40 (11/12). Neurotransmitter stimulation N (control/CIA) = day 10 (6/6), day 15 (6/3), day 20 (6/6), and day 40 (8/8) for ACh, VIP, and NA (10−6 M, 10−8 M). Data are expressed as mean ± standard error of the mean. *P < 0.05 neurotransmitter stimulation versus non-stimulated macrophages; #P < 0.05 control cells versus CIA cells. ACh acetylcholine, BMM bone marrow-derived macrophage, CIA collagen II-induced arthritis, NA noradrenaline, VIP vasoactive intestinal peptide
Mentions: Changes in apoptotic activity could also explain the reduced number of CIA BMMs (Fig. 5). (For more detailed data, see Additional file 4: Table S2E.)Fig. 5

Bottom Line: Additionally, we found a strong reduction in proliferation of BMMs at CIA onset and in the chronic phase of CIA.Apoptosis remained unaffected.We suggest an altered reactivity of BMMs to neurotransmitter stimulation caused by CIA and maybe also by aging.

View Article: PubMed Central - PubMed

Affiliation: Experimental Orthopedics, Centre for Medical Biotechnology, Biopark I, University of Regensburg, Josef-Engert-Str. 9, 93053, Regensburg, Germany. dominique.muschter@klinik.uni-regensburg.de.

ABSTRACT

Introduction: Numerous observations indicate that rheumatoid arthritis (RA) has a bone marrow component. In parallel, local synovial changes depend on neuronal components of the peripheral sympathetic nervous system. Here, we wanted to analyze whether collagen II-induced arthritis (CIA) has an impact on number, adhesion, apoptosis, and proliferation of the macrophage subset of bone marrow cells and how alterations in neurotransmitter microenvironment affect these properties.

Methods: Bone marrow-derived macrophages (BMMs) were isolated from Dark Agouti rats at different stages of CIA, and number, adhesion, caspase 3/7 activity, and proliferation were analyzed in the presence of acetylcholine (ACh), noradrenaline (NA), and vasoactive intestinal peptide (VIP).

Results: Opposed to enhanced CD11b(+) (cluster of differentiation 11b-positive) and EMR1(+) (epidermal growth factor-like module-containing mucin-like hormone receptor-like 1-positive) cells, characterizing the macrophage subset, in native bone marrow of rats with acute inflammatory arthritis, we found decreased numbers of CIA macrophages after enrichment and culture in comparison with healthy (control) animals. Adhesion studies revealed significantly reduced attachment to plastic in acute arthritis and collagen type I and fibronectin in chronic arthritis. Additionally, we found a strong reduction in proliferation of BMMs at CIA onset and in the chronic phase of CIA. Apoptosis remained unaffected. Neurotransmitter stimulation profoundly affected proliferation, adhesion, and apoptosis of BMMs from CIA and control rats, depending on disease time point. Cultured BMMs from CIA and control animals expressed neurotransmitter receptors for ACh, VIP and NA, but the expression profile seemed not to be affected by CIA.

Conclusions: Induction of CIA distinctly inhibits proliferation of BMMs in low- and non-inflammatory phases and reduces attachment to plastic at the acute inflammatory arthritis stage and adhesion to collagen I and fibronectin at the chronic stage. Influence of neurotransmitter stimulation on adhesion, apoptosis, and proliferation is altered by CIA depending on disease stage. We suggest an altered reactivity of BMMs to neurotransmitter stimulation caused by CIA and maybe also by aging.

No MeSH data available.


Related in: MedlinePlus