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The involvement of mast cells in the irinotecan-induced enteric neurons loss and reactive gliosis

View Article: PubMed Central - PubMed

ABSTRACT

Background: The irinotecan (CPT-11) causes intestinal mucositis and diarrhea that may be related to changes in the enteric nervous system (ENS). In inflammatory condition, mast cells release a variety of pro-inflammatory mediators that can interact with the ENS cells. It has not been explored whether CPT-11 is able to alter the enteric glial and neuronal cell, and the role of mast cells in this effect. Therefore, this study was conducted to investigate the effect of CPT-11 on the enteric glial and neuronal cells, as well as to study the role of mast cells in the CPT-11-induced intestinal mucositis.

Methods: Intestinal mucositis was induced in Swiss mice by the injection of CPT-11 (60 mg/kg, i.p.) once a day for 4 days following by euthanasia on the fifth day. To investigate the role of mast cells, the mice were pretreated with compound 48/80 for 4 days (first day, 0.6 mg/kg; second day, 1.0 mg/kg; third day, 1.2 mg/kg; fourth day, 2.4 mg/kg) to induce mast cell degranulation before the CPT-11 treatment.

Results: Here, we show that CPT-11 increased glial fibrillary acidic protein (GFAP) and S100β gene and S100β protein expressions and decreased HuC/D protein expression in the small intestine segments. Concomitantly, CPT-11 enhanced tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) levels and inducible nitric oxide synthase (iNOS) gene expression, associated with an increase in the total number macrophages (positive cells for ionized calcium-binding adapter molecule, Iba-1) and degranulated mast cells in the small intestine segments and caused significant weight loss. The pretreatment with compound 48/80, an inductor of mast cells degranulation, significantly prevented these CPT-11-induced effects.

Conclusions: Our data suggests the participation of mast cells on the CPT-11-induced intestinal mucositis, macrophages activation, enteric reactive gliosis, and neuron loss.

Electronic supplementary material: The online version of this article (doi:10.1186/s12974-017-0854-1) contains supplementary material, which is available to authorized users.

No MeSH data available.


Mast cell pre-degranulation prevents CPT-11-induced increase of GFAP and S100β immunostaining and reduction of HuC/D immunostaining in duodenum of mice. a Representative images illustrating GFAP, S100β, and HuC/D immunostaining in the mucosa, submucosa, and myenteric plexuses. Scale bar corresponds to 50 μm (GFAP and S100β) or 20 μm (HuC/D). bGraphs represent the mean ± SEM of the percentage of GFAP, S100β, or HuC/D positive immunostaining in duodenum related to total tissue in five (GFAP and S100β) or ten (HuC/D) microscope field per mice from six mice in each group quantified using Photoshop. #p < 0.05 versus control group. *p < 0.05 versus CPT-11 group. One-way ANOVA followed by Bonferroni
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Fig5: Mast cell pre-degranulation prevents CPT-11-induced increase of GFAP and S100β immunostaining and reduction of HuC/D immunostaining in duodenum of mice. a Representative images illustrating GFAP, S100β, and HuC/D immunostaining in the mucosa, submucosa, and myenteric plexuses. Scale bar corresponds to 50 μm (GFAP and S100β) or 20 μm (HuC/D). bGraphs represent the mean ± SEM of the percentage of GFAP, S100β, or HuC/D positive immunostaining in duodenum related to total tissue in five (GFAP and S100β) or ten (HuC/D) microscope field per mice from six mice in each group quantified using Photoshop. #p < 0.05 versus control group. *p < 0.05 versus CPT-11 group. One-way ANOVA followed by Bonferroni

Mentions: To examine whether the CPT-11-induced reactive gliosis might be mediated by mast cells, GFAP and S100β expressions were investigated by immunohistochemistry in the small intestine sections collected from mice pretreated, or not, with compound 48/80 before the CPT-11 administrations. An increased GFAP immunostaining was clearly observed in mucosa, submucosa, and myenteric plexuses of the duodenum of mice subjected to CPT-11-induced intestinal mucositis (CPT-11 group) when compared with the control group (Fig. 5a). The pretreatment with compound 48/80 (p < 0.05) prevented the increase of both GFAP and S100β-positive cells in the duodenum of the CPT-11 + c48/80 group when compared with CPT-11 group (Fig. 5a, b).Fig. 5


The involvement of mast cells in the irinotecan-induced enteric neurons loss and reactive gliosis
Mast cell pre-degranulation prevents CPT-11-induced increase of GFAP and S100β immunostaining and reduction of HuC/D immunostaining in duodenum of mice. a Representative images illustrating GFAP, S100β, and HuC/D immunostaining in the mucosa, submucosa, and myenteric plexuses. Scale bar corresponds to 50 μm (GFAP and S100β) or 20 μm (HuC/D). bGraphs represent the mean ± SEM of the percentage of GFAP, S100β, or HuC/D positive immunostaining in duodenum related to total tissue in five (GFAP and S100β) or ten (HuC/D) microscope field per mice from six mice in each group quantified using Photoshop. #p < 0.05 versus control group. *p < 0.05 versus CPT-11 group. One-way ANOVA followed by Bonferroni
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
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getmorefigures.php?uid=PMC5384042&req=5

Fig5: Mast cell pre-degranulation prevents CPT-11-induced increase of GFAP and S100β immunostaining and reduction of HuC/D immunostaining in duodenum of mice. a Representative images illustrating GFAP, S100β, and HuC/D immunostaining in the mucosa, submucosa, and myenteric plexuses. Scale bar corresponds to 50 μm (GFAP and S100β) or 20 μm (HuC/D). bGraphs represent the mean ± SEM of the percentage of GFAP, S100β, or HuC/D positive immunostaining in duodenum related to total tissue in five (GFAP and S100β) or ten (HuC/D) microscope field per mice from six mice in each group quantified using Photoshop. #p < 0.05 versus control group. *p < 0.05 versus CPT-11 group. One-way ANOVA followed by Bonferroni
Mentions: To examine whether the CPT-11-induced reactive gliosis might be mediated by mast cells, GFAP and S100β expressions were investigated by immunohistochemistry in the small intestine sections collected from mice pretreated, or not, with compound 48/80 before the CPT-11 administrations. An increased GFAP immunostaining was clearly observed in mucosa, submucosa, and myenteric plexuses of the duodenum of mice subjected to CPT-11-induced intestinal mucositis (CPT-11 group) when compared with the control group (Fig. 5a). The pretreatment with compound 48/80 (p < 0.05) prevented the increase of both GFAP and S100β-positive cells in the duodenum of the CPT-11 + c48/80 group when compared with CPT-11 group (Fig. 5a, b).Fig. 5

View Article: PubMed Central - PubMed

ABSTRACT

Background: The irinotecan (CPT-11) causes intestinal mucositis and diarrhea that may be related to changes in the enteric nervous system (ENS). In inflammatory condition, mast cells release a variety of pro-inflammatory mediators that can interact with the ENS cells. It has not been explored whether CPT-11 is able to alter the enteric glial and neuronal cell, and the role of mast cells in this effect. Therefore, this study was conducted to investigate the effect of CPT-11 on the enteric glial and neuronal cells, as well as to study the role of mast cells in the CPT-11-induced intestinal mucositis.

Methods: Intestinal mucositis was induced in Swiss mice by the injection of CPT-11 (60&nbsp;mg/kg, i.p.) once a day for 4&nbsp;days following by euthanasia on the fifth day. To investigate the role of mast cells, the mice were pretreated with compound 48/80 for 4&nbsp;days (first day, 0.6&nbsp;mg/kg; second day, 1.0&nbsp;mg/kg; third day, 1.2&nbsp;mg/kg; fourth day, 2.4&nbsp;mg/kg) to induce mast cell degranulation before the CPT-11 treatment.

Results: Here, we show that CPT-11 increased glial fibrillary acidic protein (GFAP) and S100&beta; gene and S100&beta; protein expressions and decreased HuC/D protein expression in the small intestine segments. Concomitantly, CPT-11 enhanced tumor necrosis factor-&alpha; (TNF-&alpha;) and interleukin-6 (IL-6) levels and inducible nitric oxide synthase (iNOS) gene expression, associated with an increase in the total number macrophages (positive cells for ionized calcium-binding adapter molecule, Iba-1) and degranulated mast cells in the small intestine segments and caused significant weight loss. The pretreatment with compound 48/80, an inductor of mast cells degranulation, significantly prevented these CPT-11-induced effects.

Conclusions: Our data suggests the participation of mast cells on the CPT-11-induced intestinal mucositis, macrophages activation, enteric reactive gliosis, and neuron loss.

Electronic supplementary material: The online version of this article (doi:10.1186/s12974-017-0854-1) contains supplementary material, which is available to authorized users.

No MeSH data available.