• Title/Summary/Keyword: mRNA levels

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Bacterial $\beta$-Glucan Exhibits Potent Hypoglycemic Activity via Decrease of Serum Lipids and Adiposity, and Increase of UCP mRNA Expression

  • HONG KYUNGHEE;JANG KI-HYO;LEE JAE-CHEOL;KIM SOHYE;KIM MI-KYOUNG;LEE IN-YOUNG;KIM SANG-MOO;LIM YOONG HO;KANG SOON AH
    • Journal of Microbiology and Biotechnology
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    • v.15 no.4
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    • pp.823-830
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    • 2005
  • This study was undertaken to evaluate the effect of bacteria-derived $\beta$-glucan fiber on serum lipids, adiposity and uncoupling protein (UCP) expression in rats. In order to induce obesity, Sprague-Dawley weanling male rats were allowed free access to AIN-76A diet until 4 weeks of age, and fed high-fat diet (beef tallow, $40\%$ of calories as fat) for 6 weeks until 10 weeks of age. Rats were then fed with $0\%$ thigh- fat control group), $1\%$, or $5\%$ bacterial ~-glucan supplemented high-fat diets (w/w) for another 6 weeks. For comparison, normal control group was fed with AIN-76 diet $11.7\%$ fat). Supplementation with bacterial $\beta$-glucan resulted in a significant reduction of high-fat-induced white fat (i.e., visceral and peritoneal fat) development, adipocyte hypertrophy, and development of hyperinsulinemia and hyperleptinemia. Serum triglyceride, total cholesterol, and free fatty acid levels were greatly reduced, but, HDL-cholesterol concentrations were increased by bacterial $\beta$-glucan supplementation. Serum leptin level was lower in the $\beta$-glucan groups than in the high-fat group. The expression of UCPs (UCP1, UCP2, and UCP3) in brown adipose tissue (BAT) were significantly increased by $5\%$ bacterial $\beta$-glucan-containing diet. This study suggests that the anti-obesity effect of $5\%$ bacterial $\beta$-glucan is attributed to upregulation of UCPs and inefficient energy utilization.

Baicalein Induces Programmed Cell Death in Candida albicans

  • Dai, Bao-Di;Cao, Ying-Ying;Huang, Shan;Xu, Yong-Gang;Gao, Ping-Hui;Wang, Yan;Jiang, Yuan-Ying
    • Journal of Microbiology and Biotechnology
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    • v.19 no.8
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    • pp.803-809
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    • 2009
  • Recent evidence has revealed the occurrence of an apoptotic phenotype in Candida albicans that is inducible with environmental stresses such as acetic acid, hydrogen peroxide, and amphotericin B. In the present study, we found that the Chinese herbal medicine Baicalein (BE), which was one of the skullcapflavones, can induce apoptosis in C. albicans. The apoptotic effects of BE were detected by flow cytometry using Annexin V-FITC and DAPI, and it was confirmed by transmission electron microscopy analysis. After exposure to 4 ${\mu}g$/ml BE for 12 h, about 10% of C. albicans cells were apoptotic. Both the increasing intracellular levels of reactive oxygen species (ROS) and upregulation of some redox-related genes (CAP1, SOD2, TRR1) were observed. Furthermore, we compared the survivals of CAP1 deleted, wild-type, and overexpressed strains and found that Cap1p attenuated BE-initiated cell death, which was coherent with a higher mRNA level of the CAP1 gene. In addition, the mitochondrial membrane potential of C. albicans cells changed significantly (p<0.001) upon BE treatment compared with control. Taken together, our results indicated that BE treatment induced apoptosis in C. albicans cells, and the apoptosis was associated with the breakdown of mitochondrial membrane potential.

Protective Effects of Acorn (Quercus acutissima CARR.) against IgE-mediated Allergic and Ovalbumin (OVA)-Induced Asthmatic Responses via Inhibition of Oxidative Stress

  • Chung, Mi-Ja;Jo, Hang-Soo;Choi, Ha-Na;Cho, Soo-Muk;Park, Yong-Il
    • Journal of Pharmaceutical Investigation
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    • v.41 no.6
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    • pp.355-362
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    • 2011
  • This work was performed to investigate the protective effect of ethanol extract (AEx) from acorn (Quercus acutissima CARR.) against allergic mediated responses in asthma model cells and mice. The AEx inhibited antigen-stimulated cytokine production such as interleukin (IL)-4, IL-13 and tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) and AEx also inhibited intracellular reactive oxygen species (ROS) generation against IgE-mediated allergic response in rat basophilic leukaemia RBL-2H3 cells. The ovalbumin (OVA)-sensitized mice were orally administered with AEx (100 or 300 mg/kg) and authentic tannic acid (75 mg/kg) every day for 15 days. Increased TNF-${\alpha}$ production by OVA-sensitization/challenge was significantly reduced by administration of AEx. The serum triglyceride levels of asthma mice were significantly reduced after feeding for 15 days with tannic acid or AEx. The mice fed with tannic acid or AEx also exhibited a significant reduction in body weights compared to those of asthma control group. The AEx increased the heme oxygenase (HO)-1 mRNA expression in the asthma model mice and showed DPPH radical scavenging activity. These results indicate that AEx protects against IgEmediated allergic and OVA-induced asthmatic responses via direct and indirect antioxidant activities. Reduced triglyceride and body weights may provide additional protective benefits of AEx on allergic asthma.

Relationship between Sloan-Kettering Virus Expression and Granulosa Cells of Atretic Follicles in the Rat Ovary

  • Kim, Hyun;Matsuwaki, Takashi;Yamanouchi, Keitaro;Nishihara, Masugi;Kim, Sung-Woo;Ko, Yeoung-Gyu;Yang, Boh-Suk
    • Reproductive and Developmental Biology
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    • v.35 no.3
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    • pp.341-348
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    • 2011
  • Sloan-Kettering virus gene product of a cellular protooncogene c-Ski is an unique nuclear pro-oncoprotein and belongs to the Ski/Sno proto-oncogene family. Ski plays multiple roles in a variety of cell types, it can induce both oncogenic transformation and terminal muscle differentiation when expressed at high levels. The aim of the present study was to locate Ski protein in rat ovaries in order to predict the possible involvement of Ski in follicular development and atresia. First, expression of c-Ski mRNA in the ovaries of adult female rats was confirmed by RT-PCR. Then, ovaries obtained on the day of estrus were subjected to immunohistochemical analysis for Ski and proliferating cell nuclear antigen (PCNA) in combination with terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL). Ski was expressed in granulosa cells that were positive for TUNEL, but negative for PCNA, regardless of the shape and size of follicles. Expression of Ski in TUNEL-positive granulosa cells, but not in PCNA-positive granulosa cells, was also verified in immature hypophysectomized rats having a single generation of developing and atretic follicles by treatment with equine chorionic gonadotropin (eCG). These results indicate that Ski is profoundly expressed in the granulosa cells of atretic follicles, but not in growing follicles, and suggest that Ski plays a role in apoptosis of granulosa cells during follicular atresia.

Selection of the optimal herbal composition of pomegranate concentrated powder from aqueous extracts of Eucommiae Cortex and Achyranthis Radix to treat osteoarthritis in rats

  • Choi, Beom-Rak;Ku, Sae-Kwang;Kang, Su-Jin;Park, Hye-Rim;Sung, Mi-Sun;Lee, Young-Joon;Park, Ki-Moon
    • The Journal of Korean Medicine
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    • v.38 no.4
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    • pp.11-40
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    • 2017
  • Objectives: We investigated whether a mixture of the main component of pomegranate concentrated powder (PCP) with appropriate proportions of Eucommiae Cortex (EC) and Achyranthis Radix (AR) could act synergistically as an effective treatment for osteoarthritis (OA). Methods: In order to evaluate the effects of PCP, EC, and AR against OA, knee thicknesses, maximum extension angle of each knee, anti - inflammation effects, the transcript levels of chondrogenic genes mRNA expressions in femur and tibia articular cartilage (AC) with synovial membrane (SM) were analyzed. In addition, the histopathology and immunohistochemistry of the femur and tibia AC or SM were performed. Results: The surgically-induced OA signs in rats were significantly inhibited by 28 days of continuous treatment of PCP, EC and AR single formulas, and PCP with EC:AR mixed formulas. Especially, PCP with EC:AR 4:1, 2:1 and 1:1 mixed formula treatment constantly showed significantly more favorable inhibitory activities, as compared with those of single formula of PCP, EC and AR treated rats. Conclusion: PCP and EC:AR 4:1 mixed formula showed similar OA refinement effects through potent anti-inflammatory pathways as compared with those of diclofenac treatment, and showed additional chondrocyte proliferating effects on the both femur and tibia AC.

Differential Action of trans-10, cis-12 Conjugated Linoleic Acid on Adipocyte Differentiation of Ovine and 3T3-L1 Preadipocytes

  • Iga, T.;Satoh, T.;Yamamoto, S.;Fukui, K.;Song, S.H.;Choi, K.C.;Roh, S.G.;Sasaki, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.11
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    • pp.1566-1573
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    • 2009
  • Trans-10, cis-12 conjugated linoleic acid (CLA) has been reported to inhibit the adipocyte differentiation of preadipocytes in non-ruminant animals (mice, rat, and human). However, the effects of trans-10, cis-12 CLA have not been clear in ruminants. The objective of this study was to investigate the effects of trans-10, cis-12 CLA on adipocyte differentiation of ovine preadipocytes. Differentiation of these preadipocytes was facilitated by treatment with trans-10, cis-12 CLA. Trans-10, cis-12 CLA increased the number and size of oil red O-stainable lipid drops as well as the levels of GPDH activity. PPAR-$\gamma{2}$ and adipophilin mRNA, adipogenic marker genes, were increased by treatment with trans-10, cis-12 CLA. This result was different from that observed with 3T3-L1 preadipocytes, a clonal cell line derived from rodents. Furthermore, trans-10, cis-12 CLA alone induced the adipocyte differentiation of ovine preadipocytes in differentiation-induction medium without troglitazone. These results suggest that CLA is an inducer and regulator in adipocyte differentiation of ovine preadipocytes, with species differences between ovine and rodent preadipocytes.

Cadmium-induced E-cadherin Expression in Cerebrovascular Endothelial Cells (카드뮴이 뇌혈관 내피세포에서의 E-cadherin 발현에 미치는 영향)

  • Seok, Sun-Mi;Lee, Tae-Gu;Kim, Young-Chae;Moon, Chang-Hyun;Baik, Eun-Joo;Jung, Yi-Sook;Lee, Soo-Hwan
    • Environmental Analysis Health and Toxicology
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    • v.22 no.2 s.57
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    • pp.137-145
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    • 2007
  • The effect of cadmium chloride $(CdCl_2)$ on the expression of E-cadherin was examined in bEnd.3 mouse brain endothelial cells. $CdCl_2$ induced $PGE_2$ release, which were blocked by non-steroidal antinflamatory drugs (NSAIDs) such as indomethacin and NS398 indicating the expression of COX-2 might contribute to $PGE_2$ production. $CdCl_2$ decreased the expression of E-cadherin, but not VE-cadherin at levels of mRNA and protein. Reduced expression level of E-cadherin was restored by NSAIDs, which was reversed by the addition of $PGE_2$. $CdC_2$-induced decrease of E-cadherin level was also recovered by antioxidants including N-acetylcyteine (NAC) and trolox. Together with previous report which showed $CdCl_2$ induced COX-2 expression in a cellular oxidative stress dependent manner, these data suggest that $CdCl_2$ decreases E-cadherin expression through induction of cellular oxidative stress and in turn COX-2 expression in brain endothelial cells.

Formation of Brain Tumor and Lymphoma by Deregulation of Apoptosis Related Gene Expression in VP-SV40 T Ag Transgenic Mice

  • Lee, Jeong-Woong;Lee, Eun-Ju;Lee, Hoon-Taek;Chung, Kil-Saeng;Ryoo, Zae-Yoong
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.47-47
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    • 2001
  • The neuropeptide vasopressin (VP) is a nine- amino acid hormone synthesized as preprohormone in the cell bodies of hypothalamic magnocellular neurons. The tumor in magnocellular neurons of the hypothalamus is associated with disfunctions of the cell bodies, leading to the diabetes insipidus. In order to study with the diabetes insipidus caused by a defect in VP synthesis and its secretion, we have produced the transgenic mice regulated by vasopressin promoter inserted to SV40 T antigen coding sequence (pVPSV.IGR2.1). One transgenic line expressing high levels of SV40 T antigen was propagated. The founder and all transgene positive adult animals have appeared with shorten mortality or apparent phenotypic abnormalities, including immune complex disease, and eventually die between 4 and 8 months of age. The mRNA and protein of SV40T antigen transgene were detected in brain of fetus as well as in brain, spleen, lung and lymph node in moribund at the age of 20 weeks. Histological analysis of transgenic mice showed that tumor developed in brain similar to primitive neuroectodermal tumors (PNET) in man. We also detected lymphomas in spleen and lymph node, and consequent tumor formation in various tissues of the transgenic mice. In pVPSV.IGR2.1, 21% mice showed brain tumor (PNET) at 5 weeks and 100% mice showed brain tumor after 15 weeks. In addition, Expression of apoptosis related genes (Bcl-28 & Bax) was increased over their age in mice with PNET as compared to control mice. Apoptosis related gene expression might be deregulated in mice with brain tumor. However, transgenic mice were not developed with the diabetes insipidus. These mice represent the first disease model to exhibit primitive neuroectodermal tumor in brain, as well as a unique model system for exploring the cellular pathogenesis of lymphomas.

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Novel functional roles of caspase-related genes in the regulation of apoptosis and autophagy

  • Shin, Ju-Hyun;Min, Sang-Hyun
    • The Korean Journal of Physiology and Pharmacology
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    • v.20 no.6
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    • pp.573-580
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    • 2016
  • Caspases, a family of cysteine proteases, cleave substrates and play significant roles in apoptosis, autophagy, and development. Recently, our group identified 72 genes that interact with Death Caspase-1 (DCP-1) proteins in Drosophila by genetic screening of 15,000 EP lines. However, the cellular functions and molecular mechanisms of the screened genes, such as their involvement in apoptosis and autophagy, are poorly understood in mammalian cells. In order to study the functional characterizations of the genes in human cells, we investigated 16 full-length human genes in mammalian expression vectors and tested their effects on apoptosis and autophagy in human cell lines. Our studies revealed that ALFY, BIRC4, and TAK1 induced autophagy, while SEC61A2, N-PAC, BIRC4, WIPI1, and FALZ increased apoptotic cell death. BIRC4 was involved in both autophagy and apoptosis. Western blot analysis and luciferase reporter activity indicated that ALFY, BIRC4, PDGFA, and TAK1 act in a p53-dependent manner, whereas CPSF1, SEC61A2, N-PAC, and WIPI1 appear to be p53-independent. Overexpression of BIRC4 and TAK1 caused upregulation of p53 and accumulation of its target proteins as well as an increase in p53 mRNA levels, suggesting that these genes are involved in p53 transcription and expression of its target genes followed by p53 protein accumulation. In conclusion, apoptosis and/or autophagy mediated by BIRC4 and TAK1 may be regulated by p53 and caspase activity. These novel findings may provide valuable information that will aid in a better understanding of the roles of caspase-related genes in human cell lines and be useful for the process of drug discovery.

Transdifferentiation of bovine epithelial cells towards adipocytes in the presence of myoepithelium

  • Sugathan, Subi;Lee, Sung-Jin;Shiwani, Supriya;Singh, Naresh Kumar
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.2
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    • pp.349-359
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    • 2020
  • Objective: Orchastric changes in the mammary glands are vital, especially during lactation. The secretary epithelial cells together with the supporting myoepithelial and stromal cells function cordially to secrete milk. Increase in the number of luminal epithelial cells and a decrease in adipocytes are visible during lactation, whereas the reverse happens in the involution. However, an early involution occurs if the epithelial cells transdifferentiate towards adipocytes during the lactation period. We aimed to inhibit the adipocyte transdifferentiation of luminal cells by restraining the peroxisomal proliferator-activated receptor γ (PPARγ) pathway. Methods: Linolenic acid (LA) and thiazolidinediones (TZDs) induced adipogenesis in mammary epithelial cells were conducted in monolayer, mixed culture as well as in transwell plate co-culture with mammary myoepithelial cells. Results: Co-culture with myoepithelial cells showed higher adipogenic gene expression in epithelial cells under LA+TZDs treatment. Increase in the expressions of PPARγ, CCAAT/enhancer-binding protein α and vimentin in both mRNA as well as protein levels were observed. Whereas, bisphenol A diglycidyl ether treatment blocked LA+TZDs induced adipogenesis, as it could not show a significant rise in adipose related markers. Although comparative results were found in both mixed culture and monolayer conditions, co-culture technic was found to work better than the others. Conclusion: Antagonizing PPARγ pathway in the presence of myoepithelial cells can significantly reduce the adipogenisis in epithelial cells, suggesting therapeutic inhibition of PPARγ can be considered to counter early involution or excessive adipogenesis in mammary epithelium in animals.