Deysi Viviana Tenazoa Wong

Targeted inhibition of IL‐18 attenuates irinotecan‐induced intestinal mucositis in mice

Intestinal mucositis is a common side‐effect of irinotecan‐based cancer chemotherapy regimens. This mucositis is associated with cytokine activation and NO synthesis. Production of IL‐18 is up‐regulated in patients suffering from inflammatory bowel disease. Therefore, we have investigated the role of IL‐18 in the pathogenesis of irinotecan‐induced intestinal mucositis.

Apolipoprotein E COG 133 mimetic peptide improves 5-fluorouracil-induced intestinal mucositis

BackgroundIntestinal mucositis is one of the major troublesome side effects of anticancer chemotherapy leading to poor patient compliance. In this study we addressed the role of the novel apolipoprotein E (ApoE) COG 133 mimetic peptide in 5-fluorouracil (5-FU)-challenged Swiss mice and IEC-6 cell monolayers. Experiments were also conducted in C57BL6J ApoE knock-out mice to assess the effects of apoE peptide treatment.MethodsExperimental groups were as follows: unchallenged controls, 5-FU-challenged mice (450 mg/kg, i.p) with or without the ApoE peptide (0.3, 1, and 3 μM, given twice daily i.p. for 4 days). Mice were sacrificed 3 days after 5-FU challenge. Proximal small intestinal samples were harvested for molecular biology and histological processing. We conducted ELISA assays and RT-PCR to target IL-1β, TNF-α, IL-10, iNOS, and myeloperoxidase (MPO) to assess intestinal inflammation. Cell death and NF-κB assays were also conducted in apoE knock-out mice. In our in vitro models, IEC-6 cells were exposed to 1 mM of 5-FU in glutamine free media with or without the ApoE peptide (0.02, 0.2, 2, 5, 10, and 20 μM). We investigated IEC-6 cell proliferation and migration, 24 h after the 5-FU challenge. Additionally, apoptotic IEC-6 cells were measured by Tunel and flow cytometry. Equimolar doses of the ApoA-I (D4-F) peptide were also used in some experiments for comparative studies.ResultsVillus blunting and heavy inflammatory infiltrates were seen in the 5-FU-challenged group, findings that were partially ameliorated by the ApoE peptide. We found increased intestinal MPO and pro-inflammatory IL-1β and TNF-α levels, and TNF-α and iNOS transcripts, and reduction of IL-10 following 5-FU treatment, each of which were partially abrogated by the peptide. Improvements were also found in IEC-6 cell apoptosis and migration following ApoE and D-4F treatment.ConclusionAltogether, these findings suggest that the novel ApoE COG 133 mimetic peptide can reduce 5-FU-induced intestinal changes and potentially benefit mucositis.

Interleukin-4 Modulates the Inflammatory Response in Ifosfamide-Induced Hemorrhagic Cystitis

We investigated whether interleukin-4 (IL-4) is present and capable of reducing inflammatory changes seen in ifosfamide-induced hemorrhagic cystitis. Male Swiss mice were treated with saline or ifosfamide alone or ifosfamide with the classical protocol with mesna and analyzed by changes in bladder wet weight (BWW), macroscopic and microscopic parameters, exudate, and hemoglobin quantification. In other groups, IL-4 was administered intraperitoneally 1xa0h before ifosfamide. In other experimental groups, C57BL/6 WT (wild type) and C57BL/6 WT IL-4 (−/−) knockout animals were treated with ifosfamide and analyzed for changes in BWW. Quantification of bladder IL-4 protein by ELISA in control, ifosfamide-, and mesna-treated groups was performed. Immunohistochemistry to tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β) as well as protein identification by Western blot assay for inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) was carried out on ifosfamide- and IL-4-treated animals. In other experimental groups, antiserum against IL-4 was given 30xa0min before ifosfamide. In IL-4-treated animals, the severity of hemorrhagic cystitis was significantly milder than in animals treated with ifosfamide only, an effect that was reverted with serum anti-IL-4. Moreover, knockout animals for IL-4 (−/−) exhibit a worse degree of inflammation when compared to C57BL/6 wild type. Exogenous IL-4 also attenuated TNF-α, IL-1β, iNOS, and COX-2 expressions in ifosfamide-treated bladders. IL-4, an anti-inflammatory cytokine, attenuates the inflammation seen in ifosfamide-induced hemorrhagic cystitis.

Dual effect of silymarin on experimental non-alcoholic steatohepatitis induced by irinotecan

ABSTRACT Irinotecan‐based regimens are commonly used for treatment of colorectal cancer, which is limited by mucositis and non‐alcoholic steatohepatitis (NASH). Silymarin (SIL) prevents fatty liver disease in the clinical setting and in models of liver damage induced chemically. This study investigated the possible effect of SIL on irinotecan (IRI)‐induced NASH. Swiss female mice were injected with saline (SAL 5 ml/kg i.p.), IRI (50 mg/kg i.p.), SIL (150 mg/kg p.o.) or IRI (50 mg/kg i.p.) + (SIL 1.5, 15 or 150 mg/kg p.o.) thrice/week/7 weeks. On the seventh week, blood samples were collected for transaminases assay and livers were collected for histopathology, measurement of the total lipids, malondyadehyde (MDA), non‐protein sulfhydryl groups (NPSH), cytokines (IL‐1&bgr;, IL 6 and IL‐10), 3‐nitrotyrosine (N‐Tyr) and toll‐like receptor 4 (TLR4) immunoexpression, quantification of NF‐kB, &agr;‐smooth muscle actin (&agr;‐SMA), and Escherichia coli 16S rRNA gene (RRS) expression. IRI increased liver transaminases, neutrophil infiltration, lipid accumulation, MDA, IL‐1&bgr; and IL‐6 levels, N‐Tyr and TLR4 immunostaining, NF‐kB, &agr;‐SMA expression and RRS versus the SAL group (p < 0.05). Additionally, SIL (1.5 mg/kg) improved these parameters (p < 0.05), except neutrophil infiltration and RSS versus the IRI group. Furthermore, the SIL (15 mg/kg) only improved the inflammatory parameters, the expression of &agr;‐SMA and RRS versus the IRI group (p < 0.05). The higher dose of SIL (150 mg/kg) was even more deleterious than the intermediate dose. Therefore, silymarin showed a dual effect on liver damage induced by IRI. Hepatoprotection seems to involve the inhibition of oxidative stress and protein nitrosylation, preventing activation of hepatic fibrosis mechanisms. Graphical abstract Figure. No caption available. HighlightsLow doses of silymarin attenuate irinotecan‐related liver damage.The protection involves the inhibition of inflammatory markers and oxidative stress.Silymarin also reduces bacterial accumulation in the liver and activation of TLR4.High doses of silymarin enhance irinotecan‐induced NASH and animal mortality.

Acute and neuropathic orofacial antinociceptive effect of eucalyptol

Terpenes have a wide range of pharmacological properties, including antinociceptive action. The anti-inflammatory and antinociceptive effects of eucalyptol are well established. The purpose of this study was to evaluate the antinociceptive effect of eucalyptol on acute and neuropathic orofacial pain in rodent models. Acute orofacial and corneal nociception was induced with formalin, capsaicin, glutamate and hypertonic saline in mice. In another series, animals were pretreated with capsazepine or ruthenium red to evaluate the involvement of TRPV1 receptors in the effect of eucalyptol. In a separate experiment, perinasal tissue levels of IL-1β, TNF-α and IFN-γ were measured. Rats were pretreated with eucalyptol before induction of temporomandibular joint pain with formalin or mustard oil. In another experiment, rats were submitted to infraorbital nerve transection (IONX) to induce chronic pain, followed by induction of mechanical hypersensitivity using Von Frey hairs. Locomotor performance was evaluated with the open-field test, and molecular docking was conducted on the TRPV1 channel. Pretreatment with eucalyptol significantly reduced formalin-induced nociceptive behaviors in all mouse strains, but response was more homogenous in the Swiss strain. Eucalyptol produced antinociceptive effects in all tests. The effect was sensitive to capsazepine but not to ruthenium red. Moreover, eucalyptol significantly reduced IFN-γ levels. Matching the results of the experiment in vivo, the docking study indicated an interaction between eucalyptol and TRPV1. No locomotor activity changes were observed. Our study shows that eucalyptol may be a clinically relevant aid in the treatment of orofacial pain, possibly by acting as a TRPV1 channel antagonist.

Interleukin‐18 (IL‐18) is equally expressed in inflammatory breast cancer and noninflammatory locally advanced breast cancer: A possible association with chemotherapy response

Inflammatory breast cancer (IBC) is the most aggressive form of locally advanced breast cancer. The signs of inflammation such as hyperemia and hyperthermia might suggest the possible participation of inflammatory mediators. This study investigates stromal and tumor expression of nuclear factor‐kappa B (NF‐κB) and interleukin‐18 (IL‐18) in samples obtained from IBC and noninflammatory locally advanced breast cancer (LABC) and the influence of these markers on patients prognosis.

Inhibition of neutrophil migration and reduction of oxidative stress by ethyl p-coumarate in acute and chronic inflammatory models

BACKGROUNDnIt is well known that medicinal plants and their products are relevant candidates for the treatment of inflammatory conditions. Ethyl p-coumarate is a phenylpropanoid that has similar structure to others anti-inflammatory and antioxidant substances. However, these activities have never been tested.nnnPURPOSEnThe aim of this study was to investigate the effect of ethyl p-coumarate on inflammatory and oxidative stress parameters.nnnSTUDY DESIGNnThis is an experimental study to evaluate the anti-inflammatory and antioxidant activities of ethyl p-coumarate in acute and chronic models of inflammation.nnnMETHODSnThe anti-inflammatory effect of ethyl p-coumarate was evaluated in Swiss mice by carrageenan-induced paw edema model (1%, 50xa0μl), followed by histological analysis, and edema induced by compound 48/80 (12xa0µg/paw), histamine (100 u202fµg/paw), serotonin (100xa0µg/paw) and prostaglandin E2 (3xa0nmol/paw) in comparison to indomethacin treatment (10xa0mg/kg, p.o.). In addition, peritonitis was induced by carrageenan (500xa0μg/cavity) to neutrophil and total leukocytes counting, myeloperoxidase (MPO), interleukin 6 (IL-6) and 8 (IL-8), nitrite (NO2-), glutathione (GSH) and malondialdehyde (MDA) measurements. The arthritis model was induced with Freunds complete adjuvant (id. 0.1u202fml) in female Wistar rats, with measurement of joint diameter and X-ray. Changes in gastric tissue of Swiss mice were analyzed in comparison to indomethacin (20 u202fmg/kg, p.o.).nnnRESULTSnAfter treatment with ethyl p-coumarate, the animals had no apparent toxic effects, and significantly inhibited paw edema induced by edematogenic agents, neutrophil (pu202f<u202f0.001) and total leukocyte (pu202f<u202f0.001) migration, MPO (pu202f<u202f0.01), IL-6 (pu202f<u202f0.05) and IL-8 (pu202f<u202f0.5), MDA (pu202f<u202f0.5), GSH (pu202f<u202f0.5), NO2- (pu202f<u202f0.001), joint thickness and bones changes. Furthermore, were not observed significant formation of gastric lesions.nnnCONCLUSIONnTaken together, these results suggest that ethyl p-coumarate exhibits anti-inflammatory activity through the inhibition of inflammatory mediators and leukocyte migration without causing gastric lesions.

Neutrophils contribute to the pathogenesis of hemorrhagic cystitis induced by ifosfamide

Abstract Ifosfamide (IFO) is an antineoplastic drug that is commonly used to treat gynecological and breast cancers. Hemorrhagic cystitis (HC) is a common side effect associated with IFO injection, which courses with neutrophil accumulation and affects 6–50% of patients depending on dose intensity. Here, we investigated the role of neutrophils in this inflammatory process. Female Swiss mice (n = 8/group) were injected with saline, IFO (400 mg/kg, i.p.), fucoidan (a P‐ and L‐selectins inhibitor, 100 mg/kg, i.v.) or IFO + fucoidan (1–100 mg/kg) alone or combined with mesna (80 mg/kg i.p.). Another group of mice received anti‐Ly6G antibody (500 &mgr;g/mouse, once daily for 2 days) for neutrophil depletion before IFO injection. In another experimental setting, animals received granulocyte colony‐stimulating factor (G‐CSF, 400 &mgr;g/kg), IFO (200 mg/kg), G‐CSF (25–400 &mgr;g/kg, for 5 days) + IFO (200 mg/kg, i.p.) or fucoidan + G‐CSF + IFO. Bladder injury was evaluated 12 h after IFO injection. IFO 400 mg/kg significantly increased visceral hyperalgesia, bladder edema, hemorrhage, vascular permeability, MPO, IL‐1&bgr; and IL‐6 tissue levels, and COX‐2 immunostaining and expression versus the saline group (P < 0.05). Conversely, fucoidan (100 mg/kg) significantly attenuated these parameters compared to IFO‐injected mice (P < 0.05). Additionally, fucoidan potentiated mesna protective effect when compared with IFO + mesna group (P < 0.05). Accordingly, neutrophil depletion with anti‐Ly6G reduced inflammatory parameters and bladder injury compared to IFO (P < 0.05). In contrast, G‐CSF enhanced IFO (200 mg/kg)‐induced HC, which was significantly attenuated by treatment with fucoidan (P < 0.05). Therefore, neutrophils contribute to the pathogenesis of HC. HighlightsNeutrophils contribute to the pathogenesis of hemorrhagic cystitis induced by ifosfamide.Filgastrim (a granulocyte colony stimulating factor) induces neutrophilia.Filgastrim enhances ifosfamide‐induced hemorrhagic cystitis, which is prevented by fucoidan, a P‐ and L‐selectin inhibitor.P‐ and L‐selectins are promising pharmacological targets to inhibit neutrophil migration during hemorrhagic cystitis.