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Sensory neuropathy hampers nociception-mediated bone marrow stem cell release in mice and patients with diabetes.

Dang Z, Maselli D, Spinetti G, Sangalli E, Carnelli F, Rosa F, Seganfreddo E, Canal F, Furlan A, Paccagnella A, Paiola E, Lorusso B, Specchia C, Albiero M, Cappellari R, Avogaro A, Falco A, Quaini F, Ou K, Rodriguez-Arabaolaza I, Emanueli C, Sambataro M, Fadini GP, Madeddu P - Diabetologia (2015)

Bottom Line: Patients with neuropathy showed a remarkable reduction in NK1R-HSPC mobilisation under ischaemia or upon G-CSF stimulation.Following LI, diabetic mice manifested an altered SP gradient between BM, peripheral blood and limb muscles, accompanied by a depressed recruitment of NK1R-HSPCs to the ischaemic site.Nociceptors may represent a new target for treatment of diabetic complications.

View Article: PubMed Central - PubMed

Affiliation: Bristol Heart Institute, School of Clinical Sciences, University of Bristol, Upper Maudlin Street, Bristol, BS2 8HW, UK.

ABSTRACT

Aims/hypothesis: Upon tissue injury, peripheral sensory neurons release nociceptive factors (e.g. substance P [SP]), which exert local and systemic actions including the recruitment of bone marrow (BM)-derived haematopoietic stem and progenitor cells (HSPCs) endowed with paracrine pro-angiogenic properties. We herein explore whether diabetic neuropathy interferes with these phenomena.

Methods: We first investigated the presence of sensory neuropathy in the BM of patients with type 2 diabetes by immunohistochemistry and morphometry analyses of nerve size and density and assessment of SP release by ELISA. We next analysed the association of sensory neuropathy with altered HSPC release under ischaemia or following direct stimulation with granulocyte colony-stimulating factor (G-CSF). BM and circulating HSPCs expressing the neurokinin 1 receptor (NK1R), which is the main SP receptor, were measured by flow cytometry. We finally assessed whether an altered modulation of SP secretion interferes with the mobilisation and homing of NK1R-HSPCs in a mouse model of type 2 diabetes after limb ischaemia (LI).

Results: Nociceptive fibres were reduced in the BM of patients and mice with type 2 diabetes. Patients with neuropathy showed a remarkable reduction in NK1R-HSPC mobilisation under ischaemia or upon G-CSF stimulation. Following LI, diabetic mice manifested an altered SP gradient between BM, peripheral blood and limb muscles, accompanied by a depressed recruitment of NK1R-HSPCs to the ischaemic site.

Conclusions/interpretation: Sensory neuropathy translates into defective liberation and homing of reparative HSPCs. Nociceptors may represent a new target for treatment of diabetic complications.

No MeSH data available.


Related in: MedlinePlus

Type 2 diabetes alters the levels of SP in PB and BM. (a) Immunostaining for SP in BM and graph showing SP levels in the BM of non-diabetic individuals; positive SP-stained area (ASP) as a percentage of the total area of sections (ATOT). (ND, n = 7) and patients with type 2 diabetes (n = 4, each subgroup [see (b)]) (scale bar, 50 μm). (b) Bar graphs (mean and SEM) showing quantitative analysis of immunoreactive SP in the PB of non-diabetic controls (black bar, n = 15) and patients with type 2 diabetes classified as uncomplicated (T2DM-U, white bar, n = 13) or complicated by neuropathy (T2DM-N, light grey bar, n = 35) or neuropathy and CLI (T2DM-NI, dark grey bar, n = 33). *p < 0.05, **p < 0.01 and ***p < 0.001 for indicated comparisons
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Fig2: Type 2 diabetes alters the levels of SP in PB and BM. (a) Immunostaining for SP in BM and graph showing SP levels in the BM of non-diabetic individuals; positive SP-stained area (ASP) as a percentage of the total area of sections (ATOT). (ND, n = 7) and patients with type 2 diabetes (n = 4, each subgroup [see (b)]) (scale bar, 50 μm). (b) Bar graphs (mean and SEM) showing quantitative analysis of immunoreactive SP in the PB of non-diabetic controls (black bar, n = 15) and patients with type 2 diabetes classified as uncomplicated (T2DM-U, white bar, n = 13) or complicated by neuropathy (T2DM-N, light grey bar, n = 35) or neuropathy and CLI (T2DM-NI, dark grey bar, n = 33). *p < 0.05, **p < 0.01 and ***p < 0.001 for indicated comparisons

Mentions: We first sought evidence of sensory neuropathy in the BM of patients with type 2 diabetes by conducting an analysis of neuronal fibre density on specimens collected from iliac crest biopsies or bone left over from hip reconstructive surgery. Since nerve density was similar in the two source groups, data were pooled in the final analysis. By immunohistochemistry, we recognised a striking decrease in the total number of nerve fibres that express the general neuronal marker PGP9.5 in the BM of diabetic patients with neuropathic (T2DM-N) and neuropathic/ischaemic complications (T2DM-NI) (Fig. 1a, ANOVA p < 0.001). This defect extends to nociceptive fibres that co-express PGP9.5 and SP (Fig. 1b, ANOVA p < 0.01) and to tyrosine hydroxylase-positive sympathetic fibres (ESM Fig. 1a, p < 0.05 vs non-diabetic controls). Additionally, in the BM of T2DM-N and T2DM-NI patients, both PGP9.5-positive and SP-positive nerve fibres showed a marked increase in diameter (Fig. 1c, ANOVA p < 0.0001 and p < 0.01, respectively), as well as vacuolisation and infiltration with CD68-positive macrophages (Fig. 1d). These degenerative/inflammatory features were not observed in non-diabetic controls. This association of diabetes with nociceptive fibre rarefaction and remodelling was additionally confirmed in a multivariable analysis adjusted by age, BMI, fasting glucose and HbA1c. Furthermore, neuronal rarefaction was associated with reduction in BM capillary density (ESM Fig. 1b), which identifies a distinct form of microangiopathy we have described previously in animal models and patients with diabetes [23, 24]. Furthermore, by immunofluorescence microscopy, we detected a significant reduction in SP expression in the BM of diabetic patients with neuropathy (p < 0.01 vs non-diabetic controls) but not in those with superimposed CLI (Fig. 2a). In addition, SP levels were remarkably reduced in the PB of neuropathic diabetic patients with or without CLI (p < 0.001 vs non-diabetic controls; p < 0.001 vs uncomplicated diabetes) (Fig. 2b).Fig. 1


Sensory neuropathy hampers nociception-mediated bone marrow stem cell release in mice and patients with diabetes.

Dang Z, Maselli D, Spinetti G, Sangalli E, Carnelli F, Rosa F, Seganfreddo E, Canal F, Furlan A, Paccagnella A, Paiola E, Lorusso B, Specchia C, Albiero M, Cappellari R, Avogaro A, Falco A, Quaini F, Ou K, Rodriguez-Arabaolaza I, Emanueli C, Sambataro M, Fadini GP, Madeddu P - Diabetologia (2015)

Type 2 diabetes alters the levels of SP in PB and BM. (a) Immunostaining for SP in BM and graph showing SP levels in the BM of non-diabetic individuals; positive SP-stained area (ASP) as a percentage of the total area of sections (ATOT). (ND, n = 7) and patients with type 2 diabetes (n = 4, each subgroup [see (b)]) (scale bar, 50 μm). (b) Bar graphs (mean and SEM) showing quantitative analysis of immunoreactive SP in the PB of non-diabetic controls (black bar, n = 15) and patients with type 2 diabetes classified as uncomplicated (T2DM-U, white bar, n = 13) or complicated by neuropathy (T2DM-N, light grey bar, n = 35) or neuropathy and CLI (T2DM-NI, dark grey bar, n = 33). *p < 0.05, **p < 0.01 and ***p < 0.001 for indicated comparisons
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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Fig2: Type 2 diabetes alters the levels of SP in PB and BM. (a) Immunostaining for SP in BM and graph showing SP levels in the BM of non-diabetic individuals; positive SP-stained area (ASP) as a percentage of the total area of sections (ATOT). (ND, n = 7) and patients with type 2 diabetes (n = 4, each subgroup [see (b)]) (scale bar, 50 μm). (b) Bar graphs (mean and SEM) showing quantitative analysis of immunoreactive SP in the PB of non-diabetic controls (black bar, n = 15) and patients with type 2 diabetes classified as uncomplicated (T2DM-U, white bar, n = 13) or complicated by neuropathy (T2DM-N, light grey bar, n = 35) or neuropathy and CLI (T2DM-NI, dark grey bar, n = 33). *p < 0.05, **p < 0.01 and ***p < 0.001 for indicated comparisons
Mentions: We first sought evidence of sensory neuropathy in the BM of patients with type 2 diabetes by conducting an analysis of neuronal fibre density on specimens collected from iliac crest biopsies or bone left over from hip reconstructive surgery. Since nerve density was similar in the two source groups, data were pooled in the final analysis. By immunohistochemistry, we recognised a striking decrease in the total number of nerve fibres that express the general neuronal marker PGP9.5 in the BM of diabetic patients with neuropathic (T2DM-N) and neuropathic/ischaemic complications (T2DM-NI) (Fig. 1a, ANOVA p < 0.001). This defect extends to nociceptive fibres that co-express PGP9.5 and SP (Fig. 1b, ANOVA p < 0.01) and to tyrosine hydroxylase-positive sympathetic fibres (ESM Fig. 1a, p < 0.05 vs non-diabetic controls). Additionally, in the BM of T2DM-N and T2DM-NI patients, both PGP9.5-positive and SP-positive nerve fibres showed a marked increase in diameter (Fig. 1c, ANOVA p < 0.0001 and p < 0.01, respectively), as well as vacuolisation and infiltration with CD68-positive macrophages (Fig. 1d). These degenerative/inflammatory features were not observed in non-diabetic controls. This association of diabetes with nociceptive fibre rarefaction and remodelling was additionally confirmed in a multivariable analysis adjusted by age, BMI, fasting glucose and HbA1c. Furthermore, neuronal rarefaction was associated with reduction in BM capillary density (ESM Fig. 1b), which identifies a distinct form of microangiopathy we have described previously in animal models and patients with diabetes [23, 24]. Furthermore, by immunofluorescence microscopy, we detected a significant reduction in SP expression in the BM of diabetic patients with neuropathy (p < 0.01 vs non-diabetic controls) but not in those with superimposed CLI (Fig. 2a). In addition, SP levels were remarkably reduced in the PB of neuropathic diabetic patients with or without CLI (p < 0.001 vs non-diabetic controls; p < 0.001 vs uncomplicated diabetes) (Fig. 2b).Fig. 1

Bottom Line: Patients with neuropathy showed a remarkable reduction in NK1R-HSPC mobilisation under ischaemia or upon G-CSF stimulation.Following LI, diabetic mice manifested an altered SP gradient between BM, peripheral blood and limb muscles, accompanied by a depressed recruitment of NK1R-HSPCs to the ischaemic site.Nociceptors may represent a new target for treatment of diabetic complications.

View Article: PubMed Central - PubMed

Affiliation: Bristol Heart Institute, School of Clinical Sciences, University of Bristol, Upper Maudlin Street, Bristol, BS2 8HW, UK.

ABSTRACT

Aims/hypothesis: Upon tissue injury, peripheral sensory neurons release nociceptive factors (e.g. substance P [SP]), which exert local and systemic actions including the recruitment of bone marrow (BM)-derived haematopoietic stem and progenitor cells (HSPCs) endowed with paracrine pro-angiogenic properties. We herein explore whether diabetic neuropathy interferes with these phenomena.

Methods: We first investigated the presence of sensory neuropathy in the BM of patients with type 2 diabetes by immunohistochemistry and morphometry analyses of nerve size and density and assessment of SP release by ELISA. We next analysed the association of sensory neuropathy with altered HSPC release under ischaemia or following direct stimulation with granulocyte colony-stimulating factor (G-CSF). BM and circulating HSPCs expressing the neurokinin 1 receptor (NK1R), which is the main SP receptor, were measured by flow cytometry. We finally assessed whether an altered modulation of SP secretion interferes with the mobilisation and homing of NK1R-HSPCs in a mouse model of type 2 diabetes after limb ischaemia (LI).

Results: Nociceptive fibres were reduced in the BM of patients and mice with type 2 diabetes. Patients with neuropathy showed a remarkable reduction in NK1R-HSPC mobilisation under ischaemia or upon G-CSF stimulation. Following LI, diabetic mice manifested an altered SP gradient between BM, peripheral blood and limb muscles, accompanied by a depressed recruitment of NK1R-HSPCs to the ischaemic site.

Conclusions/interpretation: Sensory neuropathy translates into defective liberation and homing of reparative HSPCs. Nociceptors may represent a new target for treatment of diabetic complications.

No MeSH data available.


Related in: MedlinePlus