• Journal of Embryo Transfer
• /
• v.17 no.3
• /
• pp.171-177
• /
• 2002

Glucocorticoid is activating mammary gland cells for lactating animals, resulting in increasing abilities of the milk synthesis. Expression of the prolactin receptor(PRL-R) in mammary gland cells was closely associated with milk production. To increase lactation ability for the Korean Native Goats at mid-lactation period. 0.05, 0.1. and 0.2 g of hydrocortisone was administrated with 5 $m\ell$ of saline. and injected into vein. For the control, 5 $m\ell$ of saline was administrated in to vein. After 24 H, the mammary gland tissue was collected, and mRNA expression rates were investigated for the alpha-casein and PRL-R using competitive PCR(polymerase chain reaction). There was no significant differences between treatment and control groups for the mRNA expression rate of PRL-R in mammary gland cells after 24 h of administration of hydrocortisone. The rate of mRNA expression for the alpha-casein was increased 37%, 630%, and 380% at 0.05, 0.1, and 0.2 g of hydrocortisone administration groups, respectively, comparing with control group. The results suggested that PR L-R mRNA expression of mammary gland cell by administration of hydrocortison was not significant, but increase of the alpha-casein mRNA expression my be differences of expression of functional proteins in the cell and expression patterns of protein secretion time to out of the cell. This study showed increase of alpha-casein mRNA expression by administration of hydrocortisone at mid-lactation period of Korean native goat.

• Development and Reproduction
• /
• v.8 no.2
• /
• pp.99-111
• /
• 2004

The present study aimed to investigate whether the expression of ADAM-8, 9, 10, 12, 15, 17 and ADAMTS-1 genes is controlled by ovarian steroid hormones. Ovariectomized mice were injected with 17 ${\beta}$-estradiol ($E_2$), progesterone ($P_4$, or $E_2+P_4$. Uterine tissues were processed for RT-PCR and immunoblotting. The results of RT-PCR showed that administration of $E_2$ increases the level of ADAM-8, 12 and ADAM17 expression compared to $P_4$ or control group. In contrast, administration of $P_4$ markedly stimulated the expression of ADAM-9, 10, 15 and ADAMTS-1, whereas $E_2$ did not. Immunoblotting analysis using anti-mouse ADAM polyclonal antibodies demonstrated that $E_2$ alone or $E_2+P_4$ treatment results in the strong expression of ADAM-8, 12 and ADAM17 proteins but $P_4$ alone or control group gave weak expression. In contrast, $P_4$ alone or $E_2$ plus $P_4$ treatment increased the expression level of ADAM-9, 10, 15 and DAMTS-1 proteins. $E_2$ alone or control group did not increase the expression. These results indicate that expression of ADAM-8, 12 and ADAM17 genes is upregulated by $E_2$ and that of ADAM-9, 10, 15 and ADAMTS-1 gene is upregulated by $P_4$.

• Journal of Animal Reproduction and Biotechnology
• /
• v.36 no.1
• /
• pp.9-16
• /
• 2021

Under the stressed condition, a complex feedback mechanism for stress is activated to maintain homeostasis of the body and secretes several stress hormones. But these stress hormones impair synthesis and secretion of the reproductive hormones, followed by suppression of ovarian function. Cytochrome P450 1A2 (CYP1A2) plays a major role in metabolizing exogenous substances and endogenous hormones, and its expression is recently identified at not only the liver but also several organs with respect to the pancreas, lung and ovary. Although the expression of CYP1A2 can be also affected by several factors, understanding for the changed pattern of the ovarian CYP1A2 expression upon stress induction is still limited. Therefore, CYP1A2 expression in the ovaries from immobilization stress-induced rats were assessed in the present study. The stress-induced rats in the present study exhibited the physiological changes in terms of increased stress hormone level and decreased body weight gains. Under immunohistological observation, the ovarian CYP1A2 expression in both control and the stressed ovary was localized in the antral to pre-ovulatory follicles. However, its expression level was significantly (p < 0.01) higher in the stress-induced group than control group. In addition, stress-induced group presented more abundant CYP1A2-positive follicles (%) than control group. Since expression of the ovarian CYP1A2 was highly related with follicle atresia, increased expression of CYP1A2 in the stressed ovary might be associated with changes of the ovarian follicular dynamics due to stress induction. We hope that these findings have important implications in the fields of the reproductive biology.

• Animal Bioscience
• /
• v.34 no.6
• /
• pp.993-1005
• /
• 2021

Objective: Zearalenone (ZEA) has estrogen-like effects. Our previous study has shown that ZEA (0.5 to 1.5 mg/kg) could induce abnormal uterine proliferation through transforming growth factor signaling pathway. To further study the other regulatory networks of uterine hypertrophy caused by ZEA, the potential mechanism of ZEA on porcine endometrial epithelial cells (PECs) was explored by the Illumina Hiseq 2000 sequencing system. Methods: The PECs were treated with ZEA at 0 (ZEA0), 5 (ZEA5), 20 (ZEA20), and 80 (ZEA80) µmol/L for 24 h. The collected cells were subjected to cell cycle, RNA-seq, real-time quantitative polymerase chain reaction, immunofluorescence, and western blot analysis. Results: The proportion of cells in the S and G2 phases decreased (p<0.05), but the proportion of cells in the G1 phase increased (p<0.05) in the ZEA80 treatment. Data analysis revealed that the expression of Wnt pathway-related genes, estrogen-related genes, and mitogen-activated protein kinase pathway-related genes increased (p<0.05), but the expression of genetic stability genes decreased (p<0.05) with increasing ZEA concentrations. The relative mRNA and protein expression of WNT1, β-catenin, glycogen synthase kinase 3β (GSK-3β) were increased (p<0.05) with ZEA increasing, while the relative mRNA and protein expression of cyclin D1 (CCND1) was decreased (p<0.05). Moreover, our immunofluorescence results indicate that β-catenin accumulated around the nucleus from the cell membrane and cytoplasm with increasing ZEA concentrations. Conclusion: In summary, ZEA can activate the Wnt/β-catenin signaling pathway by up-regulating WNT1 and β-catenin expression, to promote the proliferation and development of PECs. At the same time, the up-regulation of GSK-3β and down-regulation of CCND1, as well as the mRNA expression of other pathway related genes indicated that other potential effects of ZEA on the uterine development need further study.

• Biomedical Science Letters
• /
• v.13 no.3
• /
• pp.207-212
• /
• 2007

The purpose of this preliminary study is to improve the efficiency of gene transfer of nonviral plasmid DNA by in vivo electroporation in ischemic hindlimb muscle, tibialis anterior. Hindlimb ischemic model was aseptically made by excision of left femoral artery. Each $50\;{\mu}g$ of pEGFP-C1 and pGL3-control in $100\;{\mu}l$ 0.9% NaCl was injected in tibialis anterior muscle. In vivo electroporation was applied on the same site with 10 mm-distance 2 needle array electrodes and ECM830. In 3 groups of normal rat with different electric field strength 0, 200 and 800 V/cm, the expression of pEGFP-C1 was comparatively evaluated. In 8 groups of normal rats, the expression of pGL3-control was evaluated in 0, 40, 50, 80, 100, 150, 200 and 300 V/cm of electric field strength. In 5 groups of ischemic models, the expression of pGL3-control was analyzed on 0, 4, 7, 10 and 14 days elapsed after making ischemic models. In 9 groups of ischemic rats, the expression of pGL3-control was analyzed in the electric field strength 0, 60, 70, 80, 100, 150, 200, 250 and 300 V/cm. GFP expressions in normal tibialis anterior were high in the extent and degree in order of electric field strength of 200, 800 and 0 V/cm. Luciferase value was highest in $50{\sim}100\;V/cm$ electric field strength. In the case of ischemic models, luciferase expression was significantly increasing in the order elapsed time after making the model. The degree of luciferase expression was higher in cases of application of in vivo electroporation than in that of non-application and was highest in $100{\sim}150\;V/cm$. In conclusion, in vivo electroporation is effective in transfer and expression of plasmid DNA in normal and ischemic tibialis anterior of rat.

• Asian-Australasian Journal of Animal Sciences
• /
• v.31 no.8
• /
• pp.1141-1149
• /
• 2018

Objective: Cluster of differentiation 4 protein (CD4) gene is an important immune related gene which plays a significant role in T cell development and host resistance during viral infection. Methods: In order to unravel the relationship of CpG island methylation level of CD4 gene with its gene expression and T lymphocyte subpopulation traits, we used one typical Chinese indigenous breed (Dapulian, DP) and one commercial breed (Landrace), then predicted the CpG island of CD4 gene, determined the methylation status of CpG sites by bisulfite sequencing polymerase chain reaction (BSP), and carried out the correlation analyses of methylation frequencies of CpG sites with mRNA expression and T lymphocyte subpopulation traits. Results: There was one CpG island predicted in the upstream -2 kb region and exon one of porcine CD4 gene, which located 333 bp upstream from the start site of gene and contained nine CpG sites. The correlation analysis results indicated that the methylation frequency of CpG_2 significantly correlated with CD4 mRNA expression in the DP and Landrace combined population, though it did not reach significance level in DP and Landrace separately. Additionally, 15 potential binding transcription factors (TFs) were predicted within the CpG island, and one of them (Jumonji) contained CpG_2 site, suggesting that it may influence the CD4 gene expression through the potential binding TFs. We also found methylation frequency of CpG_2 negatively correlated with T lymphocyte subpopulation traits CD4+CD8-CD3-, CD4-CD8+CD3- and CD4+/CD8+, and positively correlated with CD4-CD8+CD3+ and CD4+CD8+CD3+ (for all correlation, p<0.01) in DP and Landrace combined population. Thus, the CpG_2 was a critical methylation site for porcine CD4 gene expression and T lymphocyte subpopulation traits. Conclusion: We speculated that increased methylation frequency of CpG_2 may lead to the decreased expression of CD4, which may have some kind of influence on T lymphocyte subpopulation traits and the immunity of DP population.

• Korean Journal of Medicinal Crop Science
• /
• v.28 no.6
• /
• pp.395-411
• /
• 2020

Background: This study investigated the anti-obesity effect of the flavonoid rich fraction (FRF) and its constituent, rutin obtained from the leaf of Morus alba L., on the lipid accumulation mechanism in 3T3-L1 adipocyte and C57BL/6 mouse models. Methods and Results: In Oil Red O staining, FRF (1,000 ㎍/㎖) treatments showed inhibition rate of 35.39% in lipid accumulation compared to that in the control. AdipoRedTM assay indicated that the triglyceride content in 3T3-L1 adipocytes treated with FRF (1,000 ㎍/㎖) was reduced to 23.22%, and free glycerol content was increased to 106.04% that of the control. FRF and its major constituent, rutin affected mRNA gene expression. Rutin contributed to the inhibition of Sterol regulatory element binding protein-1c (SREBP-1c) gene expression, and inhibited the transcription factors SREBP-1c, peroxisome proliferator-activated receptor gamma (PPAR-γ), CCAAT/enhancer binding protein α (C/EBPα), fatty acid synthase (FAS) and acetyl-CoA carboxylase (ACC). In addition, the effect of FRF administration on obesity development in C57BL/6 mice fed high-fat diet (HFD) was investigated. FRF suppressed weight gain, and reduced liver triglyceride and leptin secretion. FRF exerted potential anti-inflammatory effects by improving insulin resistance and adiponectin levels, and could thus be used to help counteract obesity. The mRNA expressions of PPAR-γ, FAS, ACC, and CPT-1 were determined in liver tissue. Quantitative real-time PCR analysis was also performed to evaluate the expression of IL-1β, IL-6, and TNF-α in epididymal adipose tissue. Compared to the control group, mice fed the HFD showed the up-regulation in PPAR-γ, FAS, IL-6, and TNF-α genes, and down-regulation in CPT1 gene expression. FRF treatement markedly reduced the expression of PPAR-γ, FAS, IL-6, and TNF-α compared to those in HFD control, whereas increased the expression level of CPT1. Conclusions: These results suggest that the FRF and its major active constituent, rutin, can be used as effective anti-obesity agents.

• The Korean Journal of Physiology and Pharmacology
• /
• v.1 no.1
• /
• pp.45-53
• /
• 1997

In humans and many animal models with chronic progressive renal diseases, angiotensin-converting enzyme (ACE) inhibitor markedly attenuates the progression of nephropathy. Several studies have reported augmented gene expression and redistribution of renal renin in partial nephrectomized rats. Although precise mechanism(s) is not known, the renin-angiotensin system (RAS) may play an important role in the progression of renal diseases. Thus, this study was undertaken to examine the gene expression of renal renin, angiotensinogen, and $AT_1$ subtypes ($AT_{1A}$ and $AT_{1B}$) in rats with diabetic nephropathy, and the influences of lipopolysaccharide (LPS)-induced septicemia on the gene expression. Four weeks after streptozotocin (STZ) treatment (55 mg/kg, i.p.), rats were randomly divided into LPS-treated (1.6 mg/kg, i.p.) and control rats. At 6 hours after LPS treatment, the rats were killed and the kidney was removed from each rat. Northern blot and reverse transcription-polymerase chain reaction (RT-PCR)techniques were used to detect mRNA expression. STZ treatment markedly attenuated body weight gain and significantly increased blood glucose level. Renal renin content (RRC) was significantly decreased in the STZ-treated rats compared to that in control rats. The renal ACE activity between STZ-treated and control rats was not significantly different. Renal renin mRNA level was prominently increased, while angiotensinogen and $AT_{1A}$ mRNA levels were slightly decreased in STZ-treated rats compared to those in controls. $AT_1$B mRNA level did not differ in both groups. Acute LPS treatment did not show any significant changes of mRNA levels of intrarenal RAS components in both groups. These results suggest that intrarenal RAS components were differentially regulated in STZ-treated diabetic rats. Further studies are required to evaluate the relationship between intrarenal RAS and other vasomodulatory systems.

• Asian Pacific Journal of Cancer Prevention
• /
• v.14 no.11
• /
• pp.6245-6248
• /
• 2013

Aims: To study effects of down-regulation of pituitary tumor-transforming gene (PTTG) on proliferation and metastasis ability of the SCL-1 cutaneous squamous cell carcinoma (CSCC) cell line and explore related mechanisms. Methods: SCL-1 cells were divided into 3 groups (untreated, siRNA control and PTTG siRNA). Cell proliferation assays were performed using a CCK-8 kit and proliferation and metastasis ability were analyzed using Boyden chambers. In addition, expression of MMP-2 and MMP-9 was detected by r-time qPCR and Western blotting. Results: Down-regulation of PTTG could markedly inhibit cell proliferation in SCL-1 cells, compared to untreated and control siRNA groups (P < 0.05). Real-time qPCR demonstrated that expression levels of PTTG, MMP-2 and MMP-9 in the PTTG siRNA group were 0.8%, 23.2% and 21.3% of untreated levels. Western blotting revealed that expression of PTTG, MMP-2 and MMP-9 proteins in the PTTG siRNA group was obviously down-regulated. The numbers of migrating cells ($51.38{\pm}4.71$) in the PTTG siRNA group was obviously lower than that in untreated group ($131.33{\pm}6.12$) and the control siRNA group ($127.72{\pm}5.20$) (P < 0.05), suggesting that decrease of proliferation and metastasis ability mediated by PTTG knock-down may be closely correlated with down-regulation of MMP-2 and MMP-9 expression. Conclusion: Inhibition of PTTG expression may be a new target for therapy of CSCC.

• The Journal of Internal Korean Medicine
• /
• v.25 no.4
• /
• pp.105-116
• /
• 2004

Objective : In this study, we used the aged bilateral common carotid artery occlusion (BCAO) rats were used to measure the effect of Bupleuri Radix (Si-Ho) on the brain ischemic injury, because aging is an important factor in storke, Method : The brian ischemic injury was induced by temporary closing of carotids on both sides in a low blood pressure state. Bupleuri Radix (Si-Ho) was orally administered in 18-month-old BCAO rats. Result : The Ischemic Damaged Hippocampus and HSP expression were analyzed by the immunohistochemical staining and the result were as follows: 1. The low numbers of pyramid cells in the hippocampus CA1 area for the ischemically injured experimental group rose to numbers simillar to those of the control group. 2. The thin neuronal cell layer in the hippocampus CA1 area for the ischemically injured experimental group returned to thickness simillar to those of the control group. 3. The normalized optical density of HSP70 expression was suppressed in CA2, DG and CA1 expression was significantly suppressed in the experimental group compared to the control group. Conclusion : These results suggested that Bupleuri Radix (Si-Ho) has a neuroprotective effect by reducing neural cell injury in the initial ischemic state.