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Selective cytoprotective effect of histamine on doxorubicin-induced hepatic and cardiac toxicity in animal models

View Article: PubMed Central - PubMed

ABSTRACT

The aim of the present work was to evaluate the potential protective effect of histamine on Doxorubicin (Dox)-induced hepatic and cardiac toxicity in different rodent species and in a triple-negative breast tumor-bearing mice model. Male Sprague Dawley rats and Balb/c mice were divided into four groups: control (received saline), histamine (5 mg/kg for rats and 1 mg/kg for mice, daily subcutaneous injection starting 24 h before treatment with Dox), Dox (2 mg/kg, intraperitoneally injected three times a week for 2 weeks) and Dox+histamine (received both treatments). Tissue toxicity was evaluated by histopathological studies and oxidative stress and biochemical parameters. The combined effect of histamine and Dox was also investigated in vitro and in vivo in human MDA-MB-231 triple-negative breast cancer model. Heart and liver of Dox-treated animals displayed severe histological damage, loss of tissue weight, increased TBARS levels and DNA damage along with an augment in serum creatine kinase-myocardial band. Pretreatment with histamine prevented Dox-induced tissue events producing a significant preservation of the integrity of both rat and mouse myocardium and liver, through the reduction of Dox-induced oxidative stress and apoptosis. Histamine treatment preserved anti-tumor activity of Dox, exhibiting differential cytotoxicity and increasing the Dox-induced inhibition of breast tumor growth. Findings provide preclinical evidence indicating that histamine could be a promising candidate as a selective cytoprotective agent for the treatment of Dox-induced cardiac and hepatic toxicity, and encourage the translation to clinical practice.

No MeSH data available.


Related in: MedlinePlus

Histamine enhances anti-proliferative properties of doxorubicin in vitro. (a) Proliferation was evaluated by the clonogenic assay in human TNBC MDA-MB-231 cells treated with Dox (0.01–10 nM) in the absence (closed circles) or presence (open circles) of 10 μM histamine. Proliferation was expressed as a percentage relative to untreated cells (n=3, *P<0.01 versus Dox; two-way ANOVA and Bonferroni post test). (b) Incorporation of BrdU, (c) TUNEL and (d) Annexin-V staining assays were evaluated in MDA-MB-231 cells that were left untreated (control; C) or were treated with histamine (HA, 10 μM) and/or doxorubicin (Dox, 10 nM) for 48 h. (e) The mRNA expression levels of p21, p27, cyclin D1 and cyclin E2 were determined 24 h after treatments using qPCR and the expression levels were normalized to the expression of β-2-microglobulin. The ΔΔCt method was used to calculate the fold change. (g) Oxidative DNA damage was evaluated by measuring 8-OHdG formation and (h) intracellular ROS levels were determined 24 h after HA and/or Dox treatments using flow cytometry. (n=3-5, *P<0.05, **P<0.01, ***P<0.001 versus control; #P<0.05, ##P<0.01 versus Dox). Time course effects of Dox and HA on (i) γH2AX (15 kDa) and (f) phospho-MAPKs (p-ERK1/2, 42/44 kDa and p-p38) were assayed by western blot. Total ERK1/2, p38 and β-actin (42 kDa) were used as loading control. Semiquantitative analyses of band intensities are shown (n=2).
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fig5: Histamine enhances anti-proliferative properties of doxorubicin in vitro. (a) Proliferation was evaluated by the clonogenic assay in human TNBC MDA-MB-231 cells treated with Dox (0.01–10 nM) in the absence (closed circles) or presence (open circles) of 10 μM histamine. Proliferation was expressed as a percentage relative to untreated cells (n=3, *P<0.01 versus Dox; two-way ANOVA and Bonferroni post test). (b) Incorporation of BrdU, (c) TUNEL and (d) Annexin-V staining assays were evaluated in MDA-MB-231 cells that were left untreated (control; C) or were treated with histamine (HA, 10 μM) and/or doxorubicin (Dox, 10 nM) for 48 h. (e) The mRNA expression levels of p21, p27, cyclin D1 and cyclin E2 were determined 24 h after treatments using qPCR and the expression levels were normalized to the expression of β-2-microglobulin. The ΔΔCt method was used to calculate the fold change. (g) Oxidative DNA damage was evaluated by measuring 8-OHdG formation and (h) intracellular ROS levels were determined 24 h after HA and/or Dox treatments using flow cytometry. (n=3-5, *P<0.05, **P<0.01, ***P<0.001 versus control; #P<0.05, ##P<0.01 versus Dox). Time course effects of Dox and HA on (i) γH2AX (15 kDa) and (f) phospho-MAPKs (p-ERK1/2, 42/44 kDa and p-p38) were assayed by western blot. Total ERK1/2, p38 and β-actin (42 kDa) were used as loading control. Semiquantitative analyses of band intensities are shown (n=2).

Mentions: To determine whether histamine could affect the anti-tumoral effect of Dox and considering that this chemotherapeutic agent is one of the first-line treatments in TNBC,27 the combined effect of histamine and Dox on proliferation in MDA-MB-231 cells was first investigated. Clonogenic assay demonstrated that both single agents induced a dose dependent inhibition on the proliferative capacity of MDA-MB-231 TNBC cells16 and histamine (10 μM) increased Dox inhibitory effect (Figure 5a). According to the calculated CI using the Chou-Talalay method,28 Dox and histamine combination showed synergistic anti-tumoral activity (CI<1) tested at a 50% effective dose, calculated at Dox (5 nM) and histamine (10 μM) (CI=0.41) or Dox (10 nM) and histamine (10 μM) combinations (CI=0.16).


Selective cytoprotective effect of histamine on doxorubicin-induced hepatic and cardiac toxicity in animal models
Histamine enhances anti-proliferative properties of doxorubicin in vitro. (a) Proliferation was evaluated by the clonogenic assay in human TNBC MDA-MB-231 cells treated with Dox (0.01–10 nM) in the absence (closed circles) or presence (open circles) of 10 μM histamine. Proliferation was expressed as a percentage relative to untreated cells (n=3, *P<0.01 versus Dox; two-way ANOVA and Bonferroni post test). (b) Incorporation of BrdU, (c) TUNEL and (d) Annexin-V staining assays were evaluated in MDA-MB-231 cells that were left untreated (control; C) or were treated with histamine (HA, 10 μM) and/or doxorubicin (Dox, 10 nM) for 48 h. (e) The mRNA expression levels of p21, p27, cyclin D1 and cyclin E2 were determined 24 h after treatments using qPCR and the expression levels were normalized to the expression of β-2-microglobulin. The ΔΔCt method was used to calculate the fold change. (g) Oxidative DNA damage was evaluated by measuring 8-OHdG formation and (h) intracellular ROS levels were determined 24 h after HA and/or Dox treatments using flow cytometry. (n=3-5, *P<0.05, **P<0.01, ***P<0.001 versus control; #P<0.05, ##P<0.01 versus Dox). Time course effects of Dox and HA on (i) γH2AX (15 kDa) and (f) phospho-MAPKs (p-ERK1/2, 42/44 kDa and p-p38) were assayed by western blot. Total ERK1/2, p38 and β-actin (42 kDa) were used as loading control. Semiquantitative analyses of band intensities are shown (n=2).
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Related In: Results  -  Collection

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fig5: Histamine enhances anti-proliferative properties of doxorubicin in vitro. (a) Proliferation was evaluated by the clonogenic assay in human TNBC MDA-MB-231 cells treated with Dox (0.01–10 nM) in the absence (closed circles) or presence (open circles) of 10 μM histamine. Proliferation was expressed as a percentage relative to untreated cells (n=3, *P<0.01 versus Dox; two-way ANOVA and Bonferroni post test). (b) Incorporation of BrdU, (c) TUNEL and (d) Annexin-V staining assays were evaluated in MDA-MB-231 cells that were left untreated (control; C) or were treated with histamine (HA, 10 μM) and/or doxorubicin (Dox, 10 nM) for 48 h. (e) The mRNA expression levels of p21, p27, cyclin D1 and cyclin E2 were determined 24 h after treatments using qPCR and the expression levels were normalized to the expression of β-2-microglobulin. The ΔΔCt method was used to calculate the fold change. (g) Oxidative DNA damage was evaluated by measuring 8-OHdG formation and (h) intracellular ROS levels were determined 24 h after HA and/or Dox treatments using flow cytometry. (n=3-5, *P<0.05, **P<0.01, ***P<0.001 versus control; #P<0.05, ##P<0.01 versus Dox). Time course effects of Dox and HA on (i) γH2AX (15 kDa) and (f) phospho-MAPKs (p-ERK1/2, 42/44 kDa and p-p38) were assayed by western blot. Total ERK1/2, p38 and β-actin (42 kDa) were used as loading control. Semiquantitative analyses of band intensities are shown (n=2).
Mentions: To determine whether histamine could affect the anti-tumoral effect of Dox and considering that this chemotherapeutic agent is one of the first-line treatments in TNBC,27 the combined effect of histamine and Dox on proliferation in MDA-MB-231 cells was first investigated. Clonogenic assay demonstrated that both single agents induced a dose dependent inhibition on the proliferative capacity of MDA-MB-231 TNBC cells16 and histamine (10 μM) increased Dox inhibitory effect (Figure 5a). According to the calculated CI using the Chou-Talalay method,28 Dox and histamine combination showed synergistic anti-tumoral activity (CI<1) tested at a 50% effective dose, calculated at Dox (5 nM) and histamine (10 μM) (CI=0.41) or Dox (10 nM) and histamine (10 μM) combinations (CI=0.16).

View Article: PubMed Central - PubMed

ABSTRACT

The aim of the present work was to evaluate the potential protective effect of histamine on Doxorubicin (Dox)-induced hepatic and cardiac toxicity in different rodent species and in a triple-negative breast tumor-bearing mice model. Male Sprague Dawley rats and Balb/c mice were divided into four groups: control (received saline), histamine (5&thinsp;mg/kg for rats and 1&thinsp;mg/kg for mice, daily subcutaneous injection starting 24&thinsp;h before treatment with Dox), Dox (2&thinsp;mg/kg, intraperitoneally injected three times a week for 2 weeks) and Dox+histamine (received both treatments). Tissue toxicity was evaluated by histopathological studies and oxidative stress and biochemical parameters. The combined effect of histamine and Dox was also investigated in vitro and in vivo in human MDA-MB-231 triple-negative breast cancer model. Heart and liver of Dox-treated animals displayed severe histological damage, loss of tissue weight, increased TBARS levels and DNA damage along with an augment in serum creatine kinase-myocardial band. Pretreatment with histamine prevented Dox-induced tissue events producing a significant preservation of the integrity of both rat and mouse myocardium and liver, through the reduction of Dox-induced oxidative stress and apoptosis. Histamine treatment preserved anti-tumor activity of Dox, exhibiting differential cytotoxicity and increasing the Dox-induced inhibition of breast tumor growth. Findings provide preclinical evidence indicating that histamine could be a promising candidate as a selective cytoprotective agent for the treatment of Dox-induced cardiac and hepatic toxicity, and encourage the translation to clinical practice.

No MeSH data available.


Related in: MedlinePlus