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

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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.


Related in: MedlinePlus

Effects of the mast cells pre-degranulation in the intestinal histopathological analysis and villus height: duodenum, jejunum, and ileum of controls (untreated); animals with CPT-11-induced intestinal mucositis; pretreated with c48/80 and submitted to CPT-11-induced intestinal mucositis or treated only with c48/80. d–f CPT-11 induces shortening of the villus (green arrows), loss of crypt architecture (brown arrows), intense inflammatory cell infiltrate (blue arrows), and cell vacuolization (black arrows) in duodenum, jejunum, and ileum. Hematoxylin and eosin; scale bar correspond to 100 μm in all figures (a–l). Segments of duodenum (m), jejunum (n), and ileum (o) were collected for measurement of villus height (ten villus/slide). Bars represent mean ± SEM of eight mice in each group. #p < 0.05 versus control group, *p < 0.05 versus CPT-11 group. One-way ANOVA followed by Bonferroni
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Fig2: Effects of the mast cells pre-degranulation in the intestinal histopathological analysis and villus height: duodenum, jejunum, and ileum of controls (untreated); animals with CPT-11-induced intestinal mucositis; pretreated with c48/80 and submitted to CPT-11-induced intestinal mucositis or treated only with c48/80. d–f CPT-11 induces shortening of the villus (green arrows), loss of crypt architecture (brown arrows), intense inflammatory cell infiltrate (blue arrows), and cell vacuolization (black arrows) in duodenum, jejunum, and ileum. Hematoxylin and eosin; scale bar correspond to 100 μm in all figures (a–l). Segments of duodenum (m), jejunum (n), and ileum (o) were collected for measurement of villus height (ten villus/slide). Bars represent mean ± SEM of eight mice in each group. #p < 0.05 versus control group, *p < 0.05 versus CPT-11 group. One-way ANOVA followed by Bonferroni

Mentions: The histopathology of the small intestine segments (duodenum, jejunum, and ileum) of animals subjected to CPT-11-induced intestinal mucositis showed severe destruction of the crypts, villus shortening (Fig. 2m–o), vacuolization of epithelial and smooth muscle cells, and intense infiltration of inflammatory cells into the mucosa, resulting in high histological scores (Fig. 2d–f; Table 2), when compared with the group not subjected to CPT-11-induced intestinal mucositis (control group; Fig. 2a–c; Table 2). The pretreatment with compound 48/80 was able to prevent the villus atrophy in the jejunum (Fig. 2n), but neither in the duodenum (Fig. 2m) nor in the ileum (Fig. 2o). Compound 48/80 also prevented the destruction of the crypts, and the infiltration of inflammatory cells into the mucosa of the three small intestine segments (Fig. 2g–i), resulting in significant differences in the histological scores (Table 2) between CPT-11 + c48/80 and CPT-11 groups observed in jejunal segments. Compound 48/80 by itself did not cause any damage to the small intestine, as illustrated by representative photomicrographs of duodenum (Fig. 2j), jejunum (Fig. 2k), and ileum (Fig. 2l) of a mouse treated with this compound. No significant differences were found between the c48/80 and the control groups (Table 2).Fig. 2


The involvement of mast cells in the irinotecan-induced enteric neurons loss and reactive gliosis
Effects of the mast cells pre-degranulation in the intestinal histopathological analysis and villus height: duodenum, jejunum, and ileum of controls (untreated); animals with CPT-11-induced intestinal mucositis; pretreated with c48/80 and submitted to CPT-11-induced intestinal mucositis or treated only with c48/80. d–f CPT-11 induces shortening of the villus (green arrows), loss of crypt architecture (brown arrows), intense inflammatory cell infiltrate (blue arrows), and cell vacuolization (black arrows) in duodenum, jejunum, and ileum. Hematoxylin and eosin; scale bar correspond to 100 μm in all figures (a–l). Segments of duodenum (m), jejunum (n), and ileum (o) were collected for measurement of villus height (ten villus/slide). Bars represent mean ± SEM of eight mice in each group. #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
Show All Figures
getmorefigures.php?uid=PMC5384042&req=5

Fig2: Effects of the mast cells pre-degranulation in the intestinal histopathological analysis and villus height: duodenum, jejunum, and ileum of controls (untreated); animals with CPT-11-induced intestinal mucositis; pretreated with c48/80 and submitted to CPT-11-induced intestinal mucositis or treated only with c48/80. d–f CPT-11 induces shortening of the villus (green arrows), loss of crypt architecture (brown arrows), intense inflammatory cell infiltrate (blue arrows), and cell vacuolization (black arrows) in duodenum, jejunum, and ileum. Hematoxylin and eosin; scale bar correspond to 100 μm in all figures (a–l). Segments of duodenum (m), jejunum (n), and ileum (o) were collected for measurement of villus height (ten villus/slide). Bars represent mean ± SEM of eight mice in each group. #p < 0.05 versus control group, *p < 0.05 versus CPT-11 group. One-way ANOVA followed by Bonferroni
Mentions: The histopathology of the small intestine segments (duodenum, jejunum, and ileum) of animals subjected to CPT-11-induced intestinal mucositis showed severe destruction of the crypts, villus shortening (Fig. 2m–o), vacuolization of epithelial and smooth muscle cells, and intense infiltration of inflammatory cells into the mucosa, resulting in high histological scores (Fig. 2d–f; Table 2), when compared with the group not subjected to CPT-11-induced intestinal mucositis (control group; Fig. 2a–c; Table 2). The pretreatment with compound 48/80 was able to prevent the villus atrophy in the jejunum (Fig. 2n), but neither in the duodenum (Fig. 2m) nor in the ileum (Fig. 2o). Compound 48/80 also prevented the destruction of the crypts, and the infiltration of inflammatory cells into the mucosa of the three small intestine segments (Fig. 2g–i), resulting in significant differences in the histological scores (Table 2) between CPT-11 + c48/80 and CPT-11 groups observed in jejunal segments. Compound 48/80 by itself did not cause any damage to the small intestine, as illustrated by representative photomicrographs of duodenum (Fig. 2j), jejunum (Fig. 2k), and ileum (Fig. 2l) of a mouse treated with this compound. No significant differences were found between the c48/80 and the control groups (Table 2).Fig. 2

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.


Related in: MedlinePlus