Protective effect of leukotriene receptor antagonist montelukast on smoking-induced lung injury in Wistar rats.

Increased activation of alveolar macrophage, neutrophil and mast cell has been proven in cigarette smoking (CS)-related lung disorders (CSLD). An increased production of cysteinyl-leukotrienes (LTs), which are mediators secreted from the mentioned cells, in response to CS has been shown in humans. The protective effect of LT1 receptor-1 antagonist (LTR-1AT) on CSLD is, however, not known. In this study we aimed to determine whether there is any protective effect of a LTR-1AT, montelukast (MK), on CSLD in Wistar rats. Nine controls and twenty-three smoke-exposed rats were enrolled into this study. Controls were exposed to non-filtered air, and the smoke-exposed rats were exposed to CS for 6 h/day, 6 days/week for three weeks. The CS-exposed rats were also treated with 0.1 mg/kg/day of MK or saline. Morphometric criteria for lung injury were determined as the mean linear intercept of alveolar septa (Lm), the volume density of alveolar septa (Vvspt) and the density of the alveolar surface area per unit volume of lung parenchyma (Sva.pa). Lung mast cells (LMC), which are a major source of LTs, were also counted. Results showed that Lm of the control group was significantly lower and Vvspt, Sva.pa of the controls were significantly higher compared to those of the CS-exposed groups. Animals treated with MK had significant protection against CSLD. Lm was significantly higher and Vvspt, Sva.pa were lower in the saline group than in the MK-treated group. The number of LMC in the CS-exposed groups was also significantly higher than that in the control group. Based on these results, one can suggest that some part of the pathogenesis of CSLD may be related to an enhanced LTs synthesis and LTR-1AT. Therefore, montelukast may protect against active or passive smoking-induced lung injury and related disorders.</p

The role of selectins in the first trimester pregnancy loss

Objective: There are no well-defined findings about reasons for first trimester abortion in some pregnancy cases. Selectins are cell adhesion proteins which are important for blastocyst implantation in the decidua. The goal of the study was to investigate the role of selectins in first trimester pregnancy loss by immunohistochemistry. Study design: Decidual and placental tissue samples have been obtained from the women with unwanted pregnancy as the control group (n=40) and missed abortion (n=40) as the study group. Immunohistochemistry technique has been used to compare P, L and E-selectin expression of the fibroblast and the decidual cells in uterine decidual stroma; and fibroblasts and mesenchymal cells in placental villous stroma. Immunostaining for P,L,E-Selectin has been evaluated semiquantitatively by HSCORE analysis. Results: Decidual cells, for E and L-selectin showed stronger staining in the study group than controls, and the difference was statistically significant (p = 0.001, p = 0.001). P-selectin showed stronger staining in the control group, but the difference was not as significant as the E and L-selectins (p=0.04). In the placenta, cytotrophoblasts and syncytiotrophoblasts showed stronger staining for P,E,L-selectins for the control group (

The Role of Proteins in Apoptosis of Somatic and Germ Cells in the Mouse Ovary

In this study, to determine the mechanisms of cell death in developing follicles, we investigated whether expression of Bcl-2, p53 and Bax play a role throughout the growth of follicles in the mice. Ovarian tissues and oocytes were obtained from 30 Mus Musculus type mouse. The immunostaining of ovarian tissue sections and oocytes was performed using anti-Bcl-2, anti-Bax and anti-p53 antibodies and terminal deoxynucleotidyl transfrase (TdT) assay (TUNEL) were also used for detection of internucleosomal DNA fragmentation. In ovarian tissue section evaluation, granulosa cells in antrum of secondary and Graaf follicles were positive with TUNEL staining. Immunoreactivity of Bcl-2 was moderate in primary and secondary follicles of granulosa cells. While immunostaining of p53 was moderate in granulosa cells of Graaf follicles, Bax immunoreactivity was moderate and strong in secondary and Graaf follicles, respectively. When granulosa cells were break off from zona pellucida, there were TUNEL positive cells. In unfertilised oocytes evaluation, TUNEL positive cells were observed in the granulosa cells which were free from zona pellucida. When the granulosa cells were attached with zona pellucida, they were TUNEL negative. While immunoreactivity of Bcl-2 was detected in both oocytes and granulosa cells which were attached with zona pellucida, immunostaining of p53 were only detected in granulosa cells which break off from zona pellucida. In conclusion, regulation of apoptosis in granulosa cells may be controlled by Bax expression and when the granulosa cells were not attached with zona pellucida, they may go into the apoptotic cascade. Therefore; we suggest that, the death of granulosa cells may control signals from intrinsic pathways in the Graaf follicule or from extrinsic pathways after ovulation. However, we consider of further studies to be necessary

The effects of pirfenidone on T helper cells in prevention of intraperitoneal adhesions

Abdominal surgery is linked with peritoneal adhesions. We investigated that the anti-fibrotic agent pirfenidone (PFD) has immune modulating activities and evaluated its effects on the function of T helper type 1 (Th1), Th2 and T regulatory (Treg) cells, which may play important roles in peritoneal adhesions. Eighteen female Wistar rats underwent right-sided parietal peritoneal and right uterine horn adhesion model. Rats were randomized into 3 groups as group 1 (control) (closure of midline abdominal incision without any agent administrations), group 2 (closure of incision after intraperitoneal administration of PFD) and group 3 (closure of incision and only oral administration of PFD for 14 days). Relaparotomy was performed 14 days after the first surgery. Effect of PFD on adhesion formation was assessed on Th1, Th2 and Treg cells counts using Anti-T-bet, Anti-GATA-3 Anti-FOXP3 antibodies respectively. Th1 counts were moderate in the control group, and didn't show a significant difference between all groups. Th2 cell counts were very high in the control group, but both intraperitoneal and oral administration of PFD resulted in a significant reduction in Th2 cell counts. Treg cell counts were low in number in the control group. In the intraperitoneal administration of PFD group, Treg cell counts were significantly lower than control group. There was no difference of the Treg cells between control groups and the oral administration of PFD group. PFD has prevention effect on intraperitoneal adhesions. This prevention effect seems to be related with the reduction in the numbers of Th2 and Treg cells

Immunohistochemical determination of mTOR pathway molecules in ovaries and uterus in rat estrous cycle stages

mTOR is a member of the PI3K/Akt/mTOR signaling pathway that participates in cell growth, proliferation, protein synthesis, transcription, angiogenesis, apoptosis and autophagy. mTOR and MAPK pahways are two important key signal pathways which are related to each other. We investigated the role of mTOR and other signaling molecules in rat ovaries and uteruses in stages of the estrous cycle. Young adult female rats were divided into four groups as proestrous, estrous, metestrous and diestrous according to vaginal smears. Immunohistochemical staining of mTORC1, IGF1, PI3K, pAKT1/2/3, ERK1 and pERK1/2 was performed and pAKT1/2/3 and ERK1 were also analyzed using western blotting on ovarian and uterine tissue samples. According to our results, PI3K/Akt/ mTOR and ERK/pERK showed an increase in the rat ovulation period. When all the groups were evaluated the immunoreactivities for all of the antibodies were detected in the oocytes, granulosa and theca cells, corpus luteum and stroma of ovary and lamina propria, surface and glandular epithelium of uterus with the strongest observed with anti-ERK1 antibody and then with a decreasing trend with anti-mTORC1, anti-pAkt1/2/3, anti-IGF1, anti-PI3K and anti-pERK1/2 antibodies in the proestrus and estrus stages. Differently from other parts of the ovary, highest antibody expression in the corpus luteum was observed in the metestrous stage. Moreover, the existence of pAKT1/2/3 and ERK1 proteins was confirmed with the Western blotting technique. We suggest that mTOR and mTOR-related ERK signaling molecules may participate in the rat ovulation process

RHAMM expression in the rat endometrium during the estrous cycle and following implantation

Background: Receptor for hyaluronic acid mediated motility (RHAMM) has intracellular and extracellular functions. In this study, we focus on the expression of RHAMM in the rat uterus during estrous cycle and implantation period. Methods: The female adult rats were divided into six groups following estrous cycle determination (n=36). The utreri of rats were collected according to estrous cycle phases (menstruation group). For the implantation groups, uteri were obtained on D4, D5 and D6 (day of implantation) of pregnancy. The tissue samples were fixed and cut into 5 μm thick sections. RHAMM was investigated using immunohistochemical techniques and the intensity of RHAMM was evaluated by using the H-score technique. Comparisons between groups were performed using Kruskal-Wallis test. Results: The RHAMM immunoreactivity of uterine antimesometrial epithelium (343.00±12.81), mesometrial subepithelium (285.00±27.26) and mesometrial stroma (270.00±36.00) were more prominent (p<0.05) in the proestrus than estrus (275.00±25.96; 220.00±14.48; 218.00±11.19) and diestrus (262.00±20.71; 192.50± 29.25; 216.00±12.97) groups, respectively. The most intense staining was seen in the epithelium on day four (275.50±30.06) and six (293.50±34.47) of pregnancy (p<0.05). Strong RHAMM expressions were in both mature and predecidual cells on D5 (256.00±18.71), (247.50±22.14) and D6 (256.00±30.72), (265.00±14.87), respectively. RHAMM expression was prominent in the nondecidual region on D5 (270.00±13.36). Conclusion: Considering the role of RHAMM in cell proliferation, differentiation and angiogenesis, spatiotemporal expression of RHAMM in the uterus during estrous cycle and peri-implantation period is a means through which uterus becomes receptive for developing an embryo

The Effects of Wortmannin and EGCG and Combined Treatments on MDA-MB-231 Breast Cancer Cell Lines via Inactivation of PI3K Signaling Pathway

Epigallocatechin gallate (EGCG), a major polyphenol in green tea, has been studied as an agent against carcinogenesis and Wortmannin is a microbial steroid and it inhibits phosphatidyl inositol 3-kinase (PI3K) pathway. The aim of this study was to investigate the effects of PI3K inhibitor Wortmannin, EGCG and combined treatments on PI3K pathway on human breast cancer cell line MDA-MB-231 using indirect immunohistochemistry method. MDA-MB-231 breast cancer cells were cultured in RPMI-1640 medium containing 10% FBS, 1% l-glutamine and 1% penicillin/streptomycin. Anti-PI3K, anti-AKT, anti-ERK, anti-NFkB, anti-c-jun and anti-EZH2 primary antibodies were used for indirect immunohistochemistry after 24 h administrations of Wortmannin (2.5 µM), EGCG (100 µM) and combination of them. The mean values of the staining intensities (mild, moderate, strong and very strong) and percentage of positively stained cells were calculated using H-Score. The results of this study showed that the combined treatment of Wortmannin and EGCG is more effective on the decreasing of immunoreactivities of PI3K pathway molecules than single administrations. The combined use of these drugs is thought to be advantageous in enhancing the development and efficacy of existing cancer treatments

An Experimental Study of Radiation Effect on Normal Tissue: Analysis of HIF-1 alpha, VEGF, eIF2, TIA-1, and TSP-1 Expression

WOS: 000331341700005PubMed: 24385827Objective: This study investigated whether or not the stress and hypoxia, which are the effects of radiation on normal vascular endothelium, leading to the release of HIF-1 alpha, VEGF, eIF2, TIA-1, and TSP-1 were related and the possibility of them stimulating angiogenesis. Materials and Methods: Twenty-four male Swiss Albino mice were separated into 4 groups. The first group was the control group (Group 1), and the second, third, and fourth groups were euthanized after 24 h (Group 2), 48 h (Group 3), and 7 days (Group 4), respectively. A single-fractioned 10 Gy of ionizing radiation was applied to all mice's pelvic zone with Co-60. Bladders were removed completely from the pelvic region. Immunohistochemistry and light microscopy were used to investigate whether there would be an increase or not in the angiogenesis pathway by using the HIF-1 alpha, VEGF, eIF2, TIA-1, and TSP-1 antibodies. Results: The HIP-1 alpha antibody showed strong staining in Group 3, while the staining intensity was less in other groups. VEGF showed weak staining in Groups 1 and 4, while moderate staining in Group 2 and strong staining in Group 3 was observed. eIF2 showed strong staining in Groups 1 and 4. Groups 2 and 3 were stained weakly. In the present study, staining with TSP-1 was very strong in the samples belonging to Group 1, while other groups showed very weak staining. Conclusion: When normal tissue was exposed to radiation, the positively effective factors (HIF-1, VEGF, eIF2, and TIA-1) on the angiogenesis pathway were increased while the negative factor (TSP-1) was decreased. Radiation may initiate physiological angiogenesis in the normal tissue and accelerate healing in the damaged normal tissue