Protection of radiation-induced damage to the hematopoietic system, small intestine and salivary glands in rats by JNJ7777120 compound, a histamine H4 ligand.

Based on previous data on the histamine radioprotective effect on highly radiosensitive tissues, in the present work we aimed at investigating the radioprotective potential of the H4R ligand, JNJ7777120, on ionizing radiation-induced injury and genotoxic damage in small intestine, salivary glands and hematopoietic tissue. For that purpose, rats were divided into 4 groups. JNJ7777120 and JNJ7777120-irradiated groups received a daily subcutaneous JNJ7777120 injection (10 mg/kg) starting 24 h before irradiation. Irradiated groups received a single dose of 5 Gy on whole-body using Cesium-137 source and were sacrificed 3 or 30 days after irradiation. Tissues were removed, fixed, stained with hematoxylin and eosin or PAS staining and histological characteristics were evaluated. Proliferative and apoptotic markers were studied by immunohistochemistry, while micronucleus assay was performed to evaluate DNA damage. Submandibular gland (SMG) function was evaluated by methacholine-induced salivation. Results indicate that JNJ7777120 treatment diminished mucosal atrophy and preserved villi and the number of crypts after radiation exposure (240±8 vs. 165±10, P<0.01). This effect was associated to a reduced apoptosis and DNA damage in intestinal crypts. JNJ7777120 reduced radiation-induced aplasia, preserving medullar components and reducing formation of micronucleus and also it accelerated bone marrow repopulation. Furthermore, it reduced micronucleus frequency in peripheral blood (27±8 vs. 149±22, in 1,000 erythrocytes, P<0.01). JNJ7777120 completely reversed radiation-induced reduced salivation, conserving glandular mass with normal histological appearance and reducing apoptosis and atrophy of SMG. JNJ7777120 exhibits radioprotective effects against radiation-induced cytotoxic and genotoxic damages in small intestine, SMG and hematopoietic tissues and, thus, could be of clinical value for patients undergoing radiotherapy

Aminoguanidine impedes human pancreatic tumor growth and metastasis development in nude mice

AIM: To study the action of aminoguanidine on pancreatic cancer xenografts in relation to cell proliferation, apoptosis, redox status and vascularization

Histamine prevents radiation-induced mesenchymal changes in breast cancer cells

Radiotherapy is a prime option for treatment of solid tumors including breast cancer though side effects are usually present. Experimental evidence shows an increase in invasiveness of several neoplastic cell types through conventional tumor irradiation. The induction of epithelial to mesenchymal transition is proposed as an underlying cause of metastasis triggered by gamma irradiation. Experiments were conducted to investigate the role of histamine on the ionizing radiation-induced epithelial to mesenchymal transition events in breast cancer cells with different invasive phenotype. We also evaluated the potential involvement of Src phosphorylation in the migratory capability of irradiated cells upon histamine treatment. MCF-7 and MDA-MB-231 mammary tumor cells were exposed to a single dose of 2 Gy of gamma radiation and five days after irradiation mesenchymal-like phenotypic changes were observed by optical microscope. The expression and subcellular localization of E-cadherin, β-catenin, vimentin and Slug were determined by immunoblot and indirect immunofluorescence. There was a decrease in the epithelial marker E-cadherin expression and an increase in the mesenchymal marker vimentin after irradiation. E-cadherin and β-catenin were mainly localized in cytoplasm. Slug positive nuclei, matrix metalloproteinase-2 activity and cell migration and invasion were significantly increased. In addition, a significant enhancement in Src phosphorylation/activation could be determined by immunoblot in irradiated cells. MCF-7 and MDA-MB-231 cells also received 1 or 20 μM histamine during 24 h previous to be irradiated. Notably, pre-treatment of breast cancer cells with 20 μM histamine prevented the mesenchymal changes induced by ionizing radiation and also reduced the migratory behavior of irradiated cells decreasing phospho-Src levels. Collectively, our results suggest that histamine may block events related to epithelial to mesenchymal transition in irradiated mammary cancer cells and open a perspective for the potential use of histamine to improve radiotherapy efficacy.Fil: Galarza, Tamara Ester. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; ArgentinaFil: Mohamad, Nora Alicia. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; ArgentinaFil: Táquez Delgado, Mónica Alejandra. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Vedoya, Guadalupe M.. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; ArgentinaFil: Crescenti, Ernesto J.. Instituto de Inmunooncología; ArgentinaFil: Bergoc, Rosa Maria. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Martin, Gabriela Adriana. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; ArgentinaFil: Cricco, Graciela Patricia. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentin

Effect of JNJ7777120 on radiation-induced morphological, proliferative and apoptotic alterations in the rat SMG.

<p>(A) SMG histopathology. (a,e) Normal histological appearance of untreated and (b,f) JNJ7777120-treated SMG (c, g). SMG of irradiated rats displaying damage in the epithelial architecture of the granular convoluted ducts, mild edema (red arrow), partial loss of eosinophilic secretor granular material and vacuoles (arrow head). (d,h) SMG of JNJ7777120-treated and irradiated animals showing preserved structure organization of secretor granules, with normal appearance of granular convoluted ducts with eosinophilic secretion. (a–d) H&E staining. (e–h) PAS staining. (i) Occasional TUNEL-positive cells in glandular duct cells in untreated and (j) JNJ777120-treated rats. (k) Massive presence of TUNEL-positive cells in ductal and acinar cells of glands of irradiated rats. (l) Significant reduction of TUNEL-positive cells in glands of treated and irradiated rats. (m,n) Similar PCNA immunoreactivity in SMG from untreated and JNJ7777120-treated rats. (o) Almost total absence of PCNA immunoreactivity in irradiated gland. (p) Partial preservation of PCNA-positive cells in treated and irradiated glands. Arrows indicate positive cells. Pictures were taken at 630x-fold magnification. Scale bar= 20 µm. (B) Average number of apoptotic cells and PCNA-positive cells are shown. Error bars represent the means ± SEM. **P<0.01, ***P<0.001 vs. Untreated; ^# P<0.05, ^{# # #} P<0.001 vs. Untreated-5Gy.</p

Effects of oligoelements Se, Zn, and Mn plus Lachesis muta venom in experimental scleroderma

Abstract: Scleroderma, sclerosis of the skin, is a severe autoimmune disease refractant to all kind of treatments. To study the in vivo effects of a combination of three oligoelements selenium (Se), zinc (Zn), and manganese (Mn) plus Lachesis muta venom (O-LM) on the bleomycin (BLM)-induced scleroderma mouse experimental model. C3H mice were randomly divided into four groups: control (phosphate-buffered saline (PBS)), O-LM, BLM, and BLM + O-LM. All administrations were performed subcutaneously into the back of mice. BLM was injected 5 days per week for three consecutive weeks and O-LM was administered simultaneously with BLM from the beginning of the experiments and lasted for 3 weeks after the final BLM or PBS injection (for O-LM and BLM + O-LM groups), when animals were sacrificed and histopathological, immunohistochemical, thiobarbituric acid reactive species (TBARS) evaluation, and autoantibodies detection were determined. O-LM significantly reduced BLM-induced enhanced dermal thickness (605 ± 47 vs. 956 ± 59 μm, P < 0.01), collagen deposition, and mast cells infiltration (43.1 ± 1.0 vs. 102 ± 14.1 mast cells, P < 0.05). O-LM administration significantly blocked BLM-induced oxidative damage and the enhanced immunoreactive fibroblasts for α-smooth muscle actin while reduced BLM-induced autoantibodies that strongly react mainly with skin and spleen. O-LM significantly reduced BLM-induced scleroderma through the modulation of antioxidant and immunological pathways

Effect of JNJ7777120 on bone marrow repopulation 30 days after whole body irradiation.

<p>(A) Bone marrow histopathology. (a,e) Normal trophism of untreated, and (b,f) JNJ7777120-treated bone marrow. (c,g) Bone marrow of irradiated rats showing a reduced number of components and an important adipose replacement (red arrow head). (d,h) Bone marrow of treated and irradiated animals demonstrating significant preservation of bone marrow components and minor adipose replacement. (a–d) H&E staining. (e–h) PCNA immunoreactivity. Arrows indicate PCNA-positive cells. 630x-fold magnification. Scale bar= 20 µm. (B) Histopathological characteristics of rat bone marrow. Error bars represent the means ± SEM (*P<0.05, **P<0.001 vs. Untreated; ^# P<0.05 vs. Untreated-5Gy).</p

Effect of JNJ7777120 on radiation-induced damage on salivary function.

<p>(A) Mean salivary secretion in irradiated and non-irradiated, untreated and JNJ7777120-treated rats. Error bars represent the means ± SEM (**P<0.01, ***P<0.001, vs. Untreated, ###P<0.001 vs. Untreated-5Gy). (B) ¶SMG’s percentage of body weight (SMG weights were divided by total body weight in grams and multiplied by 100). Data represent the means ± SEM (*P<0.05 vs. Untreated; #P<0.05 vs. Untreated-5Gy). Inset: JNJ7777120 compound significantly preserves SMG mass. (C) H₄R immunoreactivity. H₄R was detected in some (a) acini and also (b) excretory ducts. (D) AQP5 immunoreactivity. AQP5 was detected almost exclusively in acini in (a) untreated, (b) JNJ777120-treated and (d) treated and irradiated rats. (c) Reduce AQP5 immunoreactivity and altered distribution in SMG of irradiated rats. Pictures were taken at 630x-fold magnification. Scale bar= 20 µm.</p

Evidence of the radioprotective effect of JNJ7777120 on rat hematopoietic tissue 3 days after irradiation.

<p>(A) H&E stained representative bone marrow sections of (a) untreated-5Gy and (b) JNJ7777120-5Gy rats. Pictures were taken at 630x-fold magnification. Scale bar= 20 µm. (B) Micronucleus frequency. ¶ The number of micronuclei (MN) was determined in 1,000 erythrocytes and is expressed as mean ± SEM. < The number of micronuclei (MN) was determined in 1,000 bone marrow cells and is expressed as mean ± SEM (*P<0.05, **P<0.01, ***P<0.001 vs. Untreated; #P<0.05, # # #P<0.001 vs. Untreated-5Gy). (C) H&E stained representative spleen sections of (a) untreated-5Gy and (b) JNJ7777120-5Gy rats. Pictures were taken at 630x-fold magnification. Scale bar= 20 µm. (D) § Spleen’s percentage of body weight (spleen weights were divided by total body weight in grams and multiplied by 100). Data represent the means ± SEM (*P<0.05 vs. Untreated).</p

Effect of JNJ7777120 on the radiobiological parameters of two human breast cancer cell lines.

<p>(A) MDA-MB-231 and (B) MCF-7 cells were cultured in presence or absence of 10 µM JNJ7777120 and clonogenic survival was determined. (C) Radiobiological parameters (SF 2Gy, Dose 0.01, Dose 0.10) were obtained from the survival curves adjusted to the linear quadratic model [SF=e^-(αD+βD2)]. Values are means ± SEM.</p