The purpose of this study was to investigate the effect of antioxidants (vitamin E, selenium, and coenzyme $Q_{10}$) on the bleomycin-induced pulmonary lesions in male rats. Sprague-Dawley male rats were divided into 4 treatment groups ($T_1$, $T_2$, $T_3$, $T_4$) and 4 control groups ($C_1$, $C_1$, $C_3$, $C_4$). The treatment groups of rats weie given a single intratracheal dose of bleomycin (1.5 units/rat) and control groups of rats were given a single intratracheal dose of normal saline (0.15ml/rat). The rats in the $T_1$ group and $C_1$, group were dosed with normal saline (0.5ml/kg/day), the rats in the $T_2$ group and $C_2$ group were dosed with vitamin E (50mg/kg/day), the rats in the $T_3$ group and $C_3$ group were dosed with sodium selenite (3mg/kg/day) and the rats in the $T_4$ gronp and $C_4$ group were dosed with coenzyme $Q_{10}$ (2.5mg/kg/day) intraperitoneally for 7 days or 14 days, respectively. Animals were killed at 7th and 14th day after dosing with bleomycin or saline. The results obtained were as follows: 1. Lung wet weight of treatment groups of rats was increased significantly while body weight gain of them was decreased significantly in comparison with that of control groups of rats (p<0.01). 2. The ratio(%) of lung wet weight to final body weight of treatment groups of rats was increased significantly in comparison with that of control groups of rats (p<0.01). 3. The main histopathological findings of lungs observed in rats at 7th day after dosing with bleomycin were proliferation of the type II alveolar epithelial cells and fibroblasts, increased invading of macrophages into lesions, round cell infiltration and perivascular edema. 4. Lung fibrous tissues were markedly increased in rats observed at 14th day after dosing with bleomycin. 5. Pumonary lesions observed in rats dosed with bleomycin and antioxidants(vitamin E, selenium, coenzyme $Q_{10}$) were not significantly different from those of rats given bleomycin alone.
Journal of the Korean Society of Food Science and Nutrition
/
v.42
no.8
/
pp.1167-1174
/
2013
Cancer cells exhibit increased demand for glutamine-derived carbons to support anabolic processes. Indeed, the spectrum of glutamine-dependent tumors and the mechanisms through which glutamine supports cancer metabolism remain areas of active investigation. In the present study, we investigated the effects of glutamine deprivation on the correlation between tightening of tight junctions (TJs) and anti-invasive activity in human prostate carcinoma LnCap cells. Glutamine deprivation markedly inhibited cell motility and invasiveness in a time-dependent manner. The anti-invasive activity of glutamine deprivation was associated with an increased tightness of the TJ, which was demonstrated by an increase in transepithelial electrical resistance (TER). The activities of matrix metalloproteinase (MMP)-2 and MMP-9 were inhibited in a time-dependent fashion by glutamine deprivation, which was correlated with a decrease in expression of their mRNA and proteins and up-regulation of tissue inhibitors of metalloproteinases (TIMPs) expression. Furthermore, glutamine deprivation repressed the levels of the claudin family members, which are major components of TJs that play a key role in the control and selectivity of paracellular transport. Moreover, the levels of E-cadherin, a type I transmembrane glycoprotein, and snail, an epithelial to mesenchymal transition regulator and zinc finger transcription factor, were markedly modulated by glutamine deprivation. Taken together, these findings suggest that TJs and MMPs are critical targets of glutamine deprivation-induced anti-invasion in human prostate carcinoma LnCap cells.
The neurotrophins, required for the survival and differentiation of the nervous system, are known to be important for the development of the reproductive tissues. However, the signals initiating the growth of follicles, gamete development, and transport and the development of zygote in the reproductive system of cows remain ambiguous. The purpose of the present study was to identify the transcripts and proteins of Neurotrophin 4 (NT4) and its receptor tyrosine kinase B (TrkB) in bovine reproductive tissues. The transcripts and immunoreactivity of NT4 and TrkB proteins were detected by reverse transcription polymerase chain reaction and western blot analysis. Using immunohistochemistry, the specific immunoreactivity of NT4 and TrkB were detected in the oocytes of primordial follicles and in the growing primary follicles. The NT4 and TrkB immunoreactivity was predominantly observed in granulosa cells, cumulus granulosa cells, cumulus oocyte complexes, theca cells of mature follicles, as well as in the oviduct epithelial cells, uterine gland cell, and epithelium cells of the uterus during the follicular and luteal phases in cows. Expressions of NT4 and TrkB mRNAs were not significantly different among the ovary, oviduct, and uterus of the follicular phase. For the luteal phase, the expression of NT4 mRNA in the ovary was significantly higher than that in the oviduct and uterus, and the expression of TrkB mRNA in the oviduct was significantly higher than that in the ovary and uterus, as determined by fluorescence quantitative reverse transcription polymerase chain reaction. The expression of NT4 mRNA was significantly higher than that of TrkB mRNA in the ovary and uterus, whereas NT4 mRNA expression was lower than that of TrkB mRNA in the oviduct during the luteal phase. The present study hypothesizes that NT4 participates in the regulation of both gonads and extra-gonadal reproductive tissues in cows.
Gastric cancer (GC) is one of the most common malignancies in the world. It is the first cause of cancer deaths in both sexes In Iranian population. Circulating insulin-like growth factor-one (IGF-1) levels have been associated for gastric cancer. IGF-1 protein has central roles involved in the regulation of epithelial cell growth, proliferation, transformation, apoptosis and metastasis. Single nucleotide polymorphism in IGF-1 regulatory elements may lead to alter in IGF-1expression level and GC susceptibility. The aim of this study was to investigate the influence of IGF-1 gene polymorphism (rs5742612) on risk of GC and clinicopathological features for the first time in Iranian population. In total, 241 subjects including 100 patients with GC and 141 healthy controls were recruited in our study. Genotypes were analyzed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay with DNA from peripheral blood. The polymorphism was statistically analyzed to investigate the relationship with the risk of GC and clinicopathological properties. Logistic regression analysis revealed that there was no significant association between rs5742612 and the risk of GC. In addition, no significant association between genotypes and clinicopathological features was observed (p value>0.05). The frequencies of the CC, CT, and TT genotypes were 97%, 3%, and 0%, respectively, among the cases, and 97.9%, 2.1%, and 0%, respectively, among the controls. CC genotype was more frequent in cases and controls. The frequencies of C and T alleles were 98.9% and 1.1% in controls and 98.5% and 1.5% in patient respectively. Our results provide the first evidence that this variant is rare in Iranian population and it may not be a powerful genetic predisposing biomarker for prediction GC clinicopathological features in an Iranian population.
For an exact comparison of mRNA transcription in different samples or tissues with real time quantitative reverse transcription-polymerase chain reaction (qRT-PCR), it is crucial to select a suitable internal reference gene. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and beta-actin (ACTB) have been frequently considered as house-keeping genes to normalize for changes in specific gene expression. However, it has been reported that these genes are unsuitable references in some cases, because their transcription is significantly variable under particular experimental conditions and among tissues. The present study was aimed to investigate which reference genes are most suitable for the study of gastric cancer tissues using qRT-PCR. 50 pairs of gastric cancer and corresponding peritumoral tissues were obtained from patients with gastric cancer. Absolute qRT-PCR was employed to detect the expression of GAPDH, ACTB, RPII and 18sRNA in the gastric cancer samples. Comparing gastric cancer with corresponding peritumoral tissues, GAPDH, ACTB and RPII were obviously upregulated 6.49, 5.0 and 3.68 fold, respectively. Yet 18sRNA had no obvious expression change in gastric cancer tissues and the corresponding peritumoral tissues. The expression of GAPDH, ${\beta}$-actin, RPII and 18sRNA showed no obvious changes in normal gastric epithelial cells compared with gastric cancer cell lines. The carcinoembryonic antigen (CEA), a widely used clinical tumor marker, was used as a validation gene. Only when 18sRNA was used as the normalizing gene was CEA obviously elevated in gastric cancer tissues compared with peritumoral tissues. Our data show that 18sRNA is stably expressed in gastric cancer samples and corresponding peritumoral tissues. These observations confirm that there is no universal reference gene and underline the importance of specific optimization of potential reference genes for any experimental condition.
The effects of iron on gill tissue and metabolic rate represented by oxygen consumption of olive flounder, Paralichthys olivaceus were determined. The effects were further studied by means of survival rate of the fish exposed to a serial concentrations of iron. The olive flounder exposed to iron concentrations over 0.93 mg/$\ell$ showed curvature and terminal clubbing of gill lamellae at 2 weeks post-exposure. In iron concentration 4.89 mg/$\ell$, gill of the fish were seriously damaged just after 2 weeks, showing hyperplasia of filament epithelia, deformation of lamella epithelia, chloride cell damage, and separation of lamella epithelial layer, Gills exposed to 9.78 mg/$\ell$ iron concentration resulted in fusion and necrosis of the lamellae after 2 weeks. Significant decreases of metabolic rate of the fish were observed after 4 weeks at iron concentration 0,93 mg/$\ell$ and after 2 weeks at iron concentrations over 4.89 mg/$\ell$. Survival rate of the olive flounder decreased significantly after 4 weeks at the iron concentration over 4.89 mg/$\ell$. These results lead us to conclude that, as far as the iron effects are concerned, its concentrations should not exceed at least more than 0.93 mg/$\ell$ in the fish farm and coastal waters for normal growth of the olive flounder.
This study was carried out to develop knock-in vector for EGFP (enhanced green fluorescent protein) expression in porcine $\beta$-casein locus. For construction of knock-in vector using porcine $\beta$-casein gene, we cloned the $\beta$-casein genome DNA from porcine fetal fibroblast cells, EGFP and SV40 polyA signal using PCR. The knock-in vectors consisted of a 5-kb fragment as the 5' recombination arm and a 2.7-kb fragment as the 3' recombination arm. We used the neomycin resistance gene ($neo^{r}$) as a positive selectable marker and the diphtheria toxin A (DT-A) gene as a negative selectable marker. To demonstrate EGFP expression from knock-in vector, we are transfected knock-in vector that has EGFP gene in murine mammary epithelial cell line HC11 cells with pSV2 neo plasmid. The EGFP expression was detected in HC11 cells transfected knock-in vector. This result demonstrates that this knock-in vector may be used for the development of knock-in transgenic pig.
Huh Jeong Eun;Lee Hyo Jung;Song Gyu Yong;Cha Bae Cheon;Kim Han Sung;Yoo Dong Youl;Ryu Shi Yong;Kim Sung Hoon
Journal of Physiology & Pathology in Korean Medicine
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v.16
no.3
/
pp.452-457
/
2002
Galla Rhois is a gallnut of Rhus javanica Linne used for treatment of diarrhea, hemorrhage, cough, leukorrhea and toxic tumor etc in oriental medicine. For the evaluation of antitumor effect of Galla Rhois, activity based fractionation was done. We isolated an effective compound and identified 1,2,3,4,6-penta-O-galloyl-β-D-glucose(PGG) by photometric analysis such as NMR and MASS. Then, we studied the angiogenic activity of PGG. It showed a cytotoxicity against SK-OV-3, SK-OV-3, HT1080 with IC/sub 50/ of 50 ug/ml approximately. It also effectively inhibited proliferation of HUVEC cells treated by bFGF to 30% of control at 20 ug/ml and cell migration to 80% at 10 ug in a dose dependent fashion. Tube formation of HUVEC cells on matrigel was effectively suppressed from 2.5 ug/ml of concentration by PGG. Moreover, it effectively recovered the dysfunction of gap junctional intercellular communication in WB-F344 rat liver epithelial cells caused by hydrogen peroxide at 4 ug/ml suggesting it potently can inhibit tumor promotion. Taken together, it indicates 1,2,3,4,6-penta-O-galloyl- β -D-glucose has antiangiogenic activity.
Journal of Physiology & Pathology in Korean Medicine
/
v.30
no.2
/
pp.109-115
/
2016
Hataedock is a Korean medical treatment that administers herbal extracts orally to newborn infants. This method is used for alleviating harmful heat and excreting fetal wastes by meconium. The purpose of this study was to evaluate anti-inflammatory effect of Hataedock method with Douchi on 2,4-dinitrofluorobenzene (DNFB)-induced atopic dermatitis (AD). The 3-week-old NC/Nga mice were divided into 3 groups: the control group (Ctrl), the AD-induced group (AE), and the Hataedock-treated group (GT). Only the GT group was treated with Hataedock at the 3rd week. After 28 days from Hataedock treatment, we induced AD-like dermatitis to the AE and GT group by DNFB. The effects of Hataedock were evaluated by immunohistochemical method. In the epithelium, PKC-positive reaction of the GT group was decreased by 57%. In the dermal papillae, IL-4-positive reaction was decreased by 34%. In the dermis, the distribution of degranulated mast cells was decreased and substance P-positive reaction was decreased by 49%. In the skin tissue, edema was decreased and MMP-9-positive reaction was decreased by 71%. Tissue damage such as epithelial cell hyperplasia, infiltration of granulocyte and lymphocyte, and capillary distribution were also decreased. The Hataedock method with Douchi maintained skin barrier and inhibited skin-damaging factors via regulating Th2 differentiation. In conclusion, Hataedock has a potential for preventative treatment of AD. Further studies are necessary to investigate the immune-regulating mechanism and verify the safety and efficacy of the Hataedock method.
Sexually matured guppies (Poeiria reticulata) were exposed to TBTCI (0.1, 0.32, 1, 3.2, 10, 25, 32, 50, 75 and 100 $\mug/l$) for 144 hours to determine the bioaccumulation rate and effects on the reproduction and behavior. The ratio of TBT residues to $\SigmaBTs\; (TBT:\SigmaBTs)$ was 67% or higher in all the guppies exposed to TBTCl, and the higher the level of TBTCl exposed, the higher the ratio of TBT:∑BTs, suggesting that the higher the level of TBTCl exposed, the lower the metabolism rate of the fish. TBTCl exposure led to a poor reproductivity and an abnormal sexual behavior in the fish, i.e. a reduced number of the male sexual sigmoid display and of spermatophore in the efferent duct was observed in the fish exposed to 0.1 $\mug/l$ and higher levels of TBTCl, and a decreasing ratio of the testicular spermatophore cyst to the whole germ cell cysts was observed in the fish exposed to 0.32∼10 $\mug/l$)of TBTCl. The reduced ratio of the spermatophore cyst seems to be an effect of the endocrine disrupter inhibiting spermiogenesis. In the fish exposed to 25 $\mug/l$ and higher levels of TBTCl, more serious effects, such as a rapid increase of mortality, the necrosis of most of the germ cells, great damages in Sertoli cells and epithelial cells of the efferent duct, a significant increase of abnormal swimming behavior, and a cessation of feeding were observed, which suggest the acute toxicity of TBTCl inhibiting not only the reproduction and behavior but also the survival of the fish itself.
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