• Molecules and Cells
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• 제42권5호
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• pp.397-405
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• 2019

The regulatory role of long noncoding RNA (lncRNA) growth arrest-specific transcript 5 (GAS5) in both cancerous and noncancerous cells have been widely reported. This study aimed to evaluate the role of lncRNA GAS5 in heart failure caused by myocardial infarction. We reported that silence of lncRNA GAS5 attenuated hypoxia-triggered cell death, as cell viability was increased and apoptosis rate was decreased. This phenomenon was coupled with the down-regulated expression of p53, Bax and cleaved caspase-3, as well as the up-regulated expression of CyclinD1, CDK4 and Bcl-2. At the meantime, the expression of four heart failure-related miR-NAs was altered when lncRNA GAS5 was silenced (miR-21 and miR-142-5p were up-regulated; miR-30b and miR-93 were down-regulated). RNA immunoprecipitation assay results showed that lncRNA GAS5 worked as a molecular sponge for miR-142-5p. More interestingly, the protective actions of lncRNA GAS5 silence on hypoxia-stimulated cells were attenuated by miR-142-5p suppression. Besides, TP53INP1 was a target gene for miR-142-5p. Silence of lncRNA GAS5 promoted the activation of PI3K/AKT and MEK/ERK signaling pathways in a miR-142-5p-dependent manner. Collectively, this study demonstrated that silence of lncRNA GAS5 protected H9c2 cells against hypoxia-induced injury possibly via sponging miR-142-5p, functionally releasing TP53INP1 mRNA transcripts that are normally targeted by miR-142-5p.

• 대한한의학회지
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• 제22권4호
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• pp.45-57
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• 2001

Objectives : The water extract of Gwibitang (GBT) has been traditionally used for treatment of psychologic disease and brain damage in Oriental Medicine, This study was designed to investigate the effect of GBT on the glutamate-induced toxicity of rat C6 glial cells. Methods : The cultured cells were pretreated with GBT and exposed to glutamate, The cell damage was assessed by using MTT assay and Hoechst, IC-l staining, Results : GBT had protective effects in glutamate-induced cytotoxicity, which was revealed as apoptosis characterized by chromatic condensation and the loss of mitochondrial membrane potential in C6 glial cells. However, GBT and glutamate had no effect in the activation of caspase family cysteine proteases including caspase-3, -8 and -9 proteasesin C6 glial ce]]s, GBT significantly recovered the depletion of GSH and inhibited the generation of $H_2O_2$ by glutamate in C6 glial cells. In addition, both GBT and antioxidants such as GSH and NAC protected the glutamate-induced cytotoxicity in C6 glial cells, indicating that GBT possibly has antioxidative effect. Moreover, GBT also inhibited the glutamate-induced degradation of $IkB{\alpha}$ in C6 glial cells, This result suggest that GBT has some inhibitory effects on the transcriptional activation of $NF-_{k}B$. Conclusions : GBT has protective effects in glutamate-induced cytotoxicity via an antioxidative mechanism.

• 생태와환경
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• 제39권3호통권117호
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• pp.340-351
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• 2006

2004년 2월부터 2005년 2월까지 1년 동안 매 격주간 한강하류역의 6개 정점에서 식물플랑크톤군집에 미치는 물리화학적 환경요인을 조사하였다. 조사결과 수온은 $0.3{\sim}26.6^{\circ}C$, pH: 6.6${\sim}$9.1, DO: 1.89${\sim}$22.23 mg $L^{-1}$, BOD: 0.38${\sim}$9.20 mg $L^{-1}$, COD: 1.4${\sim}$15.2 mg $L^{-1}$, Conductivity: $62.5{\sim}500.0\;{\mu}s\;cm^{-1}$, SS: 3.00${\sim}$159.3 mg $L^{-1}$, chlorophyll a $1.7{\sim}71.3\;{\mu}g\;L^{-1}$ 범위로 변화하였다. 식물플랑크톤 현존량은 최저 $3.6{\times}10^2\;cells\;mL^{-1}$ (2004년 7월, 정점 3)에서 최고 $2.3{\times}10^4\;cells\;mL^{-1}$ (2005년 2월, 정점 6)까지 변화하였으며, 계절별 평균 현존량은 봄철에 $5.9{\times}10^3\;cells\;mL^{-1}$, 여름철 $2.1{\times}10^3\;cells\;mL^{-1}$, 가을철 $4.1{\times}10^3\;cells\;mL^{-1}$, 겨울철에 $8.5{\times}10^3\;cells\;mL^{-1}$로 겨울철에 가장 높았다. 식물플랑크톤 현존량에 미치는 환경요인의 영향을 규명하기 위하여, 종속변수인 현존량과 독립변수인 환경인과의 단계적 중회귀 분석을 한 결과 $R^2$=0.465였으며, 그 중 중요한 요인은 수온, COD, $NO_2-N$, $PO_4-N$, 유량, pH로 나타났다. 6개 정점을 환경요인을 매개변수로 유사도 분석을 한 결과 크게 3개 group으로 나눌 수 있었으며, 물리 ${\cdot}$ 화학적 환경요인과 생물학적 요인을 대상으로 ANOVA 분석에서는 수온, chlorophyll a, 규산염, 식물플랑크톤 현존량은 정점간 차이가 없이 하나의 group이었으며, 용존산소량, 전기전도도, COD, 아질산염, 질산염, 암모니아염, 인산염이 정점 1, 2에서 같은 group으로, 용존산소량, 전기전도도, pH, 인산염이 정점 3, 4, 5에서 같은 group으로 묶였다.

• Journal of Microbiology and Biotechnology
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• 제32권1호
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• pp.37-45
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• 2022

The fungal cell wall and membrane are the principal targets of antifungals. Herein, we report that myricetin exerts antifungal activity against Candida albicans by damaging the cell wall integrity and notably enhancing the membrane permeability. In the presence of sorbitol, an osmotic protectant, the minimum inhibitory concentration (MIC) of myricetin against C. albicans increased from 20 to 40 and 80 ㎍/ml in 24 and 72 h, respectively, demonstrating that myricetin disturbs the cell wall integrity of C. albicans. Fluorescence microscopic images showed the presence of propidium iodide-stained C. albicans cells, indicating the myricetin-induced initial damage of the cell membrane. The effects of myricetin on the membrane permeability of C. albicans cells were assessed using crystal violet-uptake and intracellular material-leakage assays. The percentage uptakes of crystal violet for myricetin-treated C. albicans cells at 1×, 2×, and 4× the MIC of myricetin were 36.5, 60.6, and 79.4%, respectively, while those for DMSO-treated C. albicans cells were 28.2, 28.9, and 29.7%, respectively. Additionally, myricetin-treated C. albicans cells showed notable DNA and protein leakage, compared with the DMSO-treated controls. Furthermore, treatment of C. albicans cells with 1× the MIC of myricetin showed a 17.2 and 28.0% reduction in the binding of the lipophilic probes diphenylhexatriene and Nile red, respectively, indicating that myricetin alters the lipid components or order in the C. albicans cell membrane, leading to increased membrane permeability. Therefore, these data will provide insights into the pharmacological worth of myricetin as a prospective antifungal for treating C. albicans infections.

• Molecules and Cells
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• 제27권5호
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• pp.533-538
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• 2009

Heat shock protein 27 (Hsp27) is a molecular chaperone protein which regulates cell apoptosis by interacting directly with the caspase activation components in the apoptotic pathways. With the assistance of the Tat protein transduction domain we directly delivered the Hsp27 into the myocardial cell line, H9c2 and demonstrate that this protein can reverse hypoxia-induced apoptosis of cells. In order to characterize the contribution of Hsp27 in blocking the two major apoptotic pathways operational within cells, we exposed H9c2 cells to staurosporine and cobalt chloride, agents that induce mitochondria-dependent (intrinsic) and -independent (extrinsic) pathways of apoptosis in cells respectively. The Tat-Hsp27 fusion protein showed a greater propensity to inhibit the effect induced by the cobalt chloride treatment. These data suggest that the Hsp27 predominantly exerts its protective effect by interfering with the components of the extrinsic pathway of apoptosis.

• 현장농수산연구지
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• 제14권1호
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• pp.107-115
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• 2012

치패 크기별 실험에서는 5mm치패의 여수율과 먹이섭취률은 각각 4.1 mL/h, 246×10⁴세포/h였으며, 15mm치패의 경우는 각각 70.8 mL/h, 4245.0×10⁴세포/h로서 치패 크기가 클수록 증가하였다. 회분외 건중량(AFDW)당 여수율 및 먹이섭취률은 7.5mm 치패에서 각각 3.3 mL/mg AFDW/h, 196.0×10⁴세포/mg AFDW/h로서 가장 높았다. 수온별 실험에서 10℃에서 여수율과 먹이섭취률은 각각 0.3×10⁴세포/h였고, 25℃에서는 각각 16.6 mL/h, 993.4×10⁴세포/h로서 25℃까지는 온도가 높을수록 여수율과 먹이섭취률이 증가하였으나, 30℃에서는 각각 12.9 mL/h, 772.6±298.6×10⁴세포/h로서 25℃보다 낮았다. 먹이생물 농도에 따라서는 여수율은 30×10⁴세포/mL일때는 21.1 mL/h였고, 농도가 240×10⁴세포/mL일 때는 2.3 mL/h로서 농도가 낮을수록 증가하였다. 먹이섭취률은 먹이생물의 농도가 60×10⁴세포/m일때 876.2×10⁴세포/h로서 최대값을 나타내었다.

• IMMUNE NETWORK
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• 제3권4호
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• pp.287-294
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• 2003

Background: In kidney transplantation, donor specific transfusion may induce tolerance as a result of some immune regulatory cells against the graft. In organ transplantation, the immune state arises from a relationship between the immunocompromised graft and the immunocompetent host. However, a reverse immunological situation exists between the graft and the host in hematopoietic stem cell transplantation (HSCT). In addition, early IL-2 injections after an allogeneic murine HSCT have been shown to prevent lethal graft versus host disease (GVHD) due to CD4+ cells. We investigated the induction of the regulatory CD4+CD25+ cells after a transfusion of irradiated recipient cells with IL-2 into a donor. Methods: The splenocytes (SP) were obtained from 6 week-old BALB/c mice ($H-2^d$) and irradiated as a single cell suspension. The donor mice (C3H/He, $H-2^k$) received $5{\times}10^6$ irradiated SP, and 5,000 IU IL-2 injected intraperitoneally on the day prior to HSCT. The CD4+CD25+ cell populations in SP treated C3H/He were analyzed. In order to determine the in vivo effect of CD4+CD25+ cells, the lethally irradiated BALB/c were transplanted with $1{\times}10^7$ donor BM and $5{\times}10^6$ CD4+CD25+ cells. The other recipient mice received either $1{\times}10^7$ donor BM with $5{\times}10^6$ CD4+ CD25- cells or the untreated SP. The survival and GVHD was assessed daily by a clinical scoring system. Results: In the MLR assay, BALB/c SP was used as a stimulator with C3H/He SP, as a responder, with or without treatment. The inhibition of proliferation was $30.0{\pm}13%$ compared to the control. In addition, the MLR with either the CD4+CD25+ or CD4+CD25- cells, which were isolated by MidiMacs, from the C3H/He SP treated with the recipient SP and IL-2 was evaluated. The donor SP treated with the recipient cells and IL-2 contained more CD4+CD25+ cells ($5.4{\pm}1.5%$) than the untreated mice SP ($1.4{\pm}0.3%$)(P<0.01). There was a profound inhibition in the CD4+CD25+ cells ($61.1{\pm}6.1%$), but a marked proliferation in the CD4+CD25- cells ($129.8{\pm}65.2%$). Mice in the CD4+CD25+ group showed low GVHD scores and a slow progression from the post-HSCT day 4 to day 9, but those in the control and CD4+CD25- groups had a high score and rapid progression (P<0.001). The probability of survival was 83.3% in the CD4+CD25+ group until post-HSC day 35 and all mice in the control and CD4+CD25- groups died on post-HSCT day 8 or 9 (P=0.0105). Conclusion: Donor graft engineering with irradiated recipient SP and IL-2 (recipient specific transfusion) can induce abundant regulatory CD4+CD25+ cells to prevent GVHD.

• 약학회지
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• 제52권3호
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• pp.212-218
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• 2008

Manganese is a naturally occurring element which is widespread in the environment. Also, manganese is an essential trace element and plays a key role in important biological reactions catalyzed by enzymes. However, exposure to high levels of manganese can cause toxicity in neurone and inhalation system, also damage in various tissues. We investigated the toxicity induced by manganese compound ($MnCl_2$) in cultured rat cardiomyocytes. Treatment of manganese to cultured cardiomyocyte led to cell death, reactive oxygen species (ROS) increase, and cytosolic caspase-3 activation. The ROS increase was related with the decreased level of glutathione. Expressions of ROS related genes such as heme oxygenase-1, thioredoxin reductase, and NADH quinone oxidase were significantly induced in manganese treated cells. These results suggest that manganese induce oxidative stress and apoptosis in cardiomyocytes, and may be the one of risk factors to cause heart dysfunction in vivo.

• BMB Reports
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• 제49권3호
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• pp.179-184
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• 2016

Bone morphogenetic protein 9 (BMP9) is a potent inducer of osteogenic differentiation of mesenchymal stem cells. The Wnt antagonist Dickkopf-1 (Dkk1) is involved in skeletal development and bone remodeling. Here, we investigated the role of Dkk1 in BMP9-induced osteogenic differentiation of MSCs. We found that overexpression of BMP9 induced Dkk1 expression in a dose-dependent manner, which was reduced by the P38 inhibitor SB203580 but not the ERK inhibitor PD98059. Moreover, Dkk1 dramatically decreased not only BMP9-induced alkaline phosphatase (ALP) activity but also the expression of osteocalcin (OCN) and osteopontin (OPN) and matrix mineralization of C3H10T1/2 cells. Furthermore, exogenous Dkk1 expression inhibited Wnt/β-catenin signaling induced by BMP9. Our findings indicate that Dkk1 negatively regulates BMP9-induced osteogenic differentiation through inhibition of the Wnt/β-catenin pathway and it could be used to optimize the therapeutic use of BMP9 and for bone tissue engineering.

• Animal cells and systems
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• 제13권3호
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• pp.283-288
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• 2009

The ubiquitous plasma membrane $Na^+/H^+$ exchanger 1 (NHE1) is rapidly activated in response to various extracellular stimuli and maintains normal cytoplasmic pH. Yeast two-hybrid screening was used in order to identify proteins interacting with NHE1 using its cytoplasmic domain as a bait from HeLa cDNA library. One of the interacting cDNA clones was human Lactate dehydrogenase B (LDHB). In vitro translated LDHB was pulled down together with GST-NHE1.cd protein in the GST pull down assay, confirming the interaction in vitro. LDHB antibody immunoprecipitated endogenous LDHB together with NHE1 from H9c2 cells, validating cellular interaction between NHE1 and LDHB. Subsequent analysis revealed that the overexpression of LDHB increased intracellular PH, implying opening of the NHE1 transporter. Moreover, overexpression of LDHB activated caspase 3 and induced cell death, consistent with the expected phenotype of hyper-activation of NHE1. Collectively, our data indicate that LDHB modulates NHE1 activity via physical interaction.