• The Korean Journal of Physiology and Pharmacology
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• 제24권2호
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• pp.173-183
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• 2020

An in vitro model for ischemia/reperfusion injury has not been well-established. We hypothesized that this failure may be caused by serum deprivation, the use of glutamine-containing media, and absence of acidosis. Cell viability of H9c2 cells was significantly decreased by serum deprivation. In this condition, reperfusion damage was not observed even after simulating severe ischemia. However, when cells were cultured under 10% dialyzed FBS, cell viability was less affected compared to cells cultured under serum deprivation and reperfusion damage was observed after hypoxia for 24 h. Reperfusion damage after glucose or glutamine deprivation under hypoxia was not significantly different from that after hypoxia only. However, with both glucose and glutamine deprivation, reperfusion damage was significantly increased. After hypoxia with lactic acidosis, reperfusion damage was comparable with that after hypoxia with glucose and glutamine deprivation. Although high-passage H9c2 cells were more resistant to reperfusion damage than low-passage cells, reperfusion damage was observed especially after hypoxia and acidosis with glucose and glutamine deprivation. Cell death induced by reperfusion after hypoxia with acidosis was not prevented by apoptosis, autophagy, or necroptosis inhibitors, but significantly decreased by ferrostatin-1, a ferroptosis inhibitor, and deferoxamine, an iron chelator. These data suggested that in our SIR model, cell death due to reperfusion injury is likely to occur via ferroptosis, which is related with ischemia/reperfusion-induced cell death in vivo. In conclusion, we established an optimal reperfusion injury model, in which ferroptotic cell death occurred by hypoxia and acidosis with or without glucose/glutamine deprivation under 10% dialyzed FBS.

• 한국의학물리학회지:의학물리
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• 제22권1호
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• pp.42-51
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• 2011

전통적으로 심근 생존능을 식별하고 심근 관류를 정확히 평가하기 위한 도구로 핵의학영상이 이용되고 있으나 경색영역을 정의하기에는 어려움이 있다. 이에 본 연구에서는 극성지도의 분포를 분석하여 특성에 맞는 적응적 임계값을 이용하여 심근경색 모델을 정량적으로 평가하고자 하였다. 쥐 심근경색 모델은 왼쪽 관상동맥을 결찰시켜 제작하였다. 소동물PET 영상은 37 MBq $^{18}F$-FDG를 쥐의 꼬리정맥에 주사한 후 60분 섭취 후 Siemens Inveon SPECT/PET 스캐너를 이용하여 20분 동안 ECG 신호와 함께 획득하였고, OSEM 2D 알고리즘을 이용하여 재구성하였다. PET 영상의 심근 극성지도는 Siemens QGS 소프트웨어에 적합한 형식으로 변환 후 자동으로 심근 벽을 설정하여 작성하였다. 심근경색영역의 기준데이터는 TTC 염색으로 설정하였으며 전체 좌심실대비 염색된 영역의 백분율로 획득하였다. 최적의 임계값 설정을 위해 절대치 설정 방법, Otsu 알고리즘, 다중가우시안혼합모델(Multi Gaussian mixture model, MGMM)을 이용하여 평가하였다. 절대치 설정 방법은 10~90%까지 10%단위로 미리 정의 된 임계값을 이용하였고, Otsu 알고리즘은 영상 내에서 두 군집의 분산을 최대로 하는 임계값으로 설정하였다. MGMM 방법은 영상의 화소 강도를 분석하여 여러 개의 가우시안 분포함수(MGMM2, $\cdots$ MGMM4)로 반복 수행하여 최적의 가우시안 분포를 구하여 적응적 임계값을 설정하였다. 극성지도 평가지표는 각각의 알고리즘에서 측정된 임계값을 이용하여 이진화하고 전체 극성지도와 경색영역의 백분율로 획득한 후, TTC 염색으로 획득된 기준데이터와의 차이를 비교하였다. 그 차이는 절대치 방법의 20%에서 $7.04{\pm}3.44%$, 30%에서 $3.87{\pm}2.09%$, 40%에서 $2.15{\pm}2.07%$이었다. Otsu 방법은 $3.56{\pm}4.16%$이었으며 MGMM 방법은 $2.29{\pm}1.94%$이었다. 소동물 PET 극성지도에서는 30% 임계값이 조직학적 데이터와 비교하여 가장 작은 차이를 보였다. 그러나 TTC 염색으로 측정한 크기가 10% 이하에서는 MGMM 방법이 절대치 방법보다 작은 차이를 보였다(MGMM: 0.006%, 절대치방법: 0.59%). 이 연구에서는 심근경색 모델 평가를 위하여 생체영상 극성지도에서 다중가우시안혼합모델을 이용하여 평가하고자 하였다. MGMM은 사용자의 선택 없이도 자동적으로 영상 특성을 고려하여 적응적 임계값을 찾아주는 방법으로 극성지도에서 심근경색을 평가하는데 도움이 될 것으로 기대된다.

• 대한핵의학회지
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• 제32권6호
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• pp.497-508
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• 1998

목적: 휴식기 T1-201/디피리다몰 부하 게이트 Tc-99m-MIBI/24시간 T1-201 SPECT을 이용하여 심근 벽운동의 수술 후 호전 가능성을 예측하고 어떤 지표가 예측률이 좋은지 조사하였다. 대상 및 방법: 39명(남자:여자=34:5, 나이: $58{\pm}8세$)에서 우회로 수술을 시행하고 수술전과 수술한지 3개월 후에 심근 관류 SPECT를 하여 수술 전 SPECT로부터 부하-휴식기 가역성, 휴식기 T1-201 섭취, T1-201 휴식-재분포 양상, 심근의 수축기 두꺼워짐을 준정량적으로 등급화하고 이 지표가 수술 후 심근 벽운동 호전여부를 얼마나 잘 예측할 수 있는지 보았다. 휴식기에 관류 감소가 있는 16명은 24시간 지연재분포 T1-201 SPECT를 촬영하였다. 17분절로 나누어 관류는 0에서 3 (0: 정상, 1: 가벼운 감소, 2 심한 감소, 3: 결손), 벽운동은 0 에서 4 (0: 정상, 1. 가벼운 저운동, 2: 심한 저운동, 3: 무운동, 4: 이상운동), 심근의 수축기 두꺼워짐은 좋거나 나쁨으로 판정하였다. 전체 99동맥영역의 585분절 중에서 142분절이 벽운동이 이상이 있어서 우회로 또는 수술적 성형술로 재관류 수술을 시행하였다. 결과: 수술 후 구혈률은 수술 전에 구혈률이 낮은 환자 22명은 $37.8{\pm}9.0%$에 비해 $45.5{\pm}12.3%$ 까지 증가하였다. 103 개의 분절(72.5%)의 벽운동이 수술 후 호전되었다. 부하-휴식기 가역성, 휴식기 T1-201 섭취, T1-201 휴식-재분포 양상, 심근의 수축기 두꺼워짐의 벽운동 호전 예측능은 각각 83%, 76%, 43%, 69%이었다. 음성예측률은 48%, 44%, 58%, 21%이었다. 네 지표중 어느 하나라도 있는 경우 양성예측률은 74% 음성 예측률은 46%이었다. 판단도표 분석에 의한 양성예측률은 78%, 음성예측률은 58%이었다. 단변량분석에서 부하-휴식기 가역성(p=0.0008)과 휴식기 T1-201 섭취(p=0.024)가 유의한 지표였으나 다변량 단계별 로짓분석에서는 부하-휴식기 가역성(p=0.0008)만 유의하였다. 결론 휴식기 운동이상 분절의 심근생존능을 조사한 이 연구에서 휴식기 T1-201/디피리다몰 부하 게이트 Tc-99m-MIBI/24시간 T1-201 SPECT에서 얻은 여러 지표로 수술 후 벽운동 호전을 예측할 수 있지만 부하-휴식기 가역성이 생존 심근을 찾는데 유용한 실제적인 예후 지표임을 알았다. 환자 단위로 수술여부를 판단할 때 생존심근을 가진 환자를 찾을 때에도 부하-휴식기 가역성이 중요한 단일지표인지 조사할 필요가 있다고 생각하였다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권23호
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• pp.10407-10412
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• 2015

Background: ${\beta}$-elemene, extracted from herb medicine Curcuma wenyujin has potent anti-tumor effects in various cancer cell lines. However, the activity of ${\beta}$-elemene against glioma cells remains unclear. In the present study, we assessed effects of ${\beta}$-elemene on human glioma cells and explored the underlying mechanism. Materials and Methods: Human glioma U87 cells were used. Cell proliferation was determined with MTT assay and colony formation assay to detect the effect of ${\beta}$-elemene at different doses and times. Fluorescence microscopy was used to observe cell apoptosis with Hoechst 33258 staining and change of glioma apoptosis and cell cycling were analyzed by flow cytometry. Real-time quantitative PCR and Western-blotting assay were performed to investigated the influence of ${\beta}$-elemene on expression levels of Fas/FasL, caspase-3, Bcl-2 and Bax. The experiment was divided into two groups: the blank control group and ${\beta}$-elemne treatment group. Results: With increase in the concentration of ${\beta}$-elemene, cytotoxic effects were enhanced in the glioma cell line and the concentration of inhibited cell viability ($IC_{50}$) was $48.5{\mu}g/mL$ for 24h. ${\beta}$-elemene could induce cell cycle arrest in the G0/G1 phase. With Hoechst 33258 staining, apoptotic nuclear morphological changes were observed. Activation of caspase-3,-8 and -9 was increased and the pro-apoptotic factors Fas/FasL and Bax were upregulated, while the anti-apoptotic Bcl-2 was downregulated after treatment with ${\beta}$-elemene at both mRNA and protein levels. Furthermore, proliferation and colony formation by U87 cells were inhibited by ${\beta}$-elemene in a time and does-dependent manner. Conclusions: Our results indicate that ${\beta}$-elemene inhibits growth and induces apoptosis of human glioma cells in vitro. The induction of apoptosis appears to be related with the upregulation of Fas/FasL and Bax, activation of caspase-3,-8 and -9 and downregulation of Bcl-2, which then trigger major apoptotic cascades.

• 대한핵의학회지
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• 제30권1호
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• pp.112-117
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• 1996

휴식 T1-20l/부하 Tc-99m MIBI 지연 T1-201 심근 SPECT는 관상동맥질환을 진단하는데 87%의 진단 정확성을 보여 기존의 부하/휴식 Tc-99m MIBI SPECT와 비교하여 비슷하거나 더 좋은 성능을 보였다. 이 연구로써 휴식 T1-201/부하 Tc-99m MIBI 심근 SPECT 방법의 관동맥질환 진단성능은 다른 방법들과 비슷함을 확인하였다. 또한 이 방법은 진정한 의미의 휴식기 영상을 얻을 수 있고 휴식기와 부하기 영상사이에 남아있던 방사능으로 인한 오차에서 벗어날 수 있으며 T1-201의 재분포영상과 24시간 지연영상으로부터 지속관류감소 부위의 생존여부에 대한 정보를 얻을 수 있고 휴식기 촬영후 바로 부하기 촬영을 하므로 전체 검사시간이 줄어 환자에게 편리하고 효율이 높아, 지연 T1-201 SPECT의 심근 생존능 판별성능이 확인되면 관동맥질환의 진단과 생존심근을 찾는 검사로 유용할 가능성이 있음을 알았다.

• BMB Reports
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• 제45권12호
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• pp.742-747
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• 2012

Granulocyte colony-stimulating factor (G-CSF) is used for heart failure therapy and promotes myocardial regeneration by inducing mobilization of bone marrow stem cells to the injured heart after myocardial infarction; however, this treatment has one weakness in that its biological effect is transient. In our previous report, we generated 5 mutants harboring N-linked glycosylation to improve its antiapoptotic activities. Among them, one mutant (Phe140Asn) had higher cell viability than wild-type hG-CSF in rat cardiomyocytes, even after treatment with an apoptotic agent ($H_2O_2$). Cells treated with this mutant significantly upregulated the antiapoptotic proteins, and experienced reductions in caspase 3 activity and PARP cleavage. Moreover, the total number of apoptotic cells was dramatically lower in cultures treated with mutant hG-CSF. Taken together, these results suggest that the addition of an N-linked glycosylation was successful in improving the antiapoptotic activity of hG-CSF, and that this mutated product will be a feasible therapy for patients who have experienced heart failure.

• 대한치과마취과학회지
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• 제14권3호
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• pp.157-165
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• 2014

Background: Incisional site of surgical operation become transient ischemic state and then occur reoxygenation due to vasodilatation by inflammatory reaction, the productive reactive oxygen species (ROS) give rise to many physiologic results. Apoptosis have major role on elimination of inflammatory cell and formation of granulation tissue in normal wound healing process. Remifentanil can prevent the inflammatory response and can suppress inducible nitric oxide synthase expression in a septic mouse model. After cardiopulmonary bypass for coronary artery surgery, remifentanil can also inhibit the release of biomarkers of myocardial damage. Here we investigated whether remifentanil pretreatment has cellular protective effect against hypoxia-reoxygenation in HaCaT human keratinocytes, if so, the role of apoptosis and autophagy on this phenomenon. Methods: The HaCaT human keratinocytes were exposed to various concentrations of remifentanil (0.01, 0.05, 0.1, 0.5 and 1 ng/ml) for 2 h before hypoxia (RPC/HR group). These cells were cultured under 1% oxygen tension for 24h at $37^{\circ}C$. After hypoxia, to simulate reoxygenation and recovery, the cells were reoxygenated for 12 h at $37^{\circ}C$. 3-MA/RPC/HR group was treated 3-methyladenine (3-MA), autophagy inhibitor for 1h before remifentanil treatment. Cell viability was measured using a quantitative colorimetric assay with thiazolyl blue tetrazoliumbromide (MTT, amresco), showing the mitochondrial activity of living cells. To investigate whether the occurrence of autophagy and apoptosis, we used fluorescence microscopy and Western blot analysis. Results: The viability against hypoxia-reoxygenation injury in remifentanil preconditioning keratinocytes were increased, and these cells were showed stimulated expression of autophagy 3-MA suppressed the induction of autophagy effectively and the protective effects on apoptosis. Atg5, Beclin-1, LC3-II and p62 were elevated in RPC/HR group. But they were decreased when autophagy was suppressed by 3-MA. Conclusions: Remifentanil preconditioning showed the protective effect in human keratinocytes, and we concluded that autophagy may take the major role in the recovery of wound from hypoxia-reoxygenation injury. We suggest that further research is needed about the cell protective effects of autophagy.

• 대한한의학회지
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• 제29권4호
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• pp.171-179
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• 2008

Objective: Bufonis Venenum is the traditional Korean medicine Chan Su, which is obtained from the skin and parotid venom gland of the toads. It has been used for myocardial diseases, inflammation diseases, pain relief, cancer and others. The main components of BV are cinobufotoxin, cinobufalin, bufalin and others. Of these, bufalin, the major active ingredient of BV, has been reported to induce apoptosis and to possess anti-tumor effects. There was no report of anti-tumor screening of BV on hepatic cancer and which signaling pathway can be involved. In order to examine the effect of BV on hepatic cancer and the related signaling pathway with BV-induced apoptosis, human Hep G2 cells were used. Methods: Analysis of apoptosis was confirmed by MTT assay. BV decreased cell viability in a dose and duration dependent manner. To observe which signaling molecules will be activated by BV, phosphorylation of MAPK (p38, ERK, JNK), caspase 8 and caspase 9 were examined by Western blot analysis. Results: The phosphorylation levels of p38 started to increase at 5 min after addition of 5 ${\mu}g$/ml of BV and sustained to increase until 48 hours. The phosphorylation levels of other MAPK (ERK and JNK), caspase 8 and caspase 9 increased in a time-dependent manner. These imply that BV may activate different signaling pathways, MAPK, caspase 8 and caspase 9. These results propose that BV may induce apoptosis on Hep G2 cells through the activation of MAPK, caspase 8 and caspase 9.

• 대한핵의학회지
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• 제26권2호
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• pp.360-364
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• 1992

Treatment for the brain tumors consist of surgery, chemotherapy, and a variety of methods of irradiation. Therapy is aimed to destroy the tumor, but necrosis and edema occur concurrently. Conventional structural imaging techniques such as CT or MRI are unable to reliably distinguish persistent and recurrent tumor from necrosis or edema. T1-201 has been shown to be useful in the evaluation of the myocardial viability by comparing the early uptake and redistribution image. The aim of this study is to evaluate the clinical usefulness of the early uptake and delayed washout images of the T1-201 brain SPECT in the brain tumors. In the pathologically diagnosed various brain tumor patients, brain SPECT was done with rotating gamma camera 15 minutes and 3 hours after T1-201 injection, and the T1-201 uptake in the tumor was compared with the skull and scalp activity. In the glioblastoma multiforme, meningioma and metastatic tumor, the T1-201 uptake was higher than low grade glioma in both 15 minute and 3 hour images (p<0.02). In the low grade glioma,3 hour T1-201 uptake was significantly lower than 15 minute uptake (p<0.05) but in the glioblastoma, meningioma and metastatic tumor there was no significant difference. There was no significant difference in the T1-201 uptake among the glioblastoma, meningioma and metastatic tumors. In one matastatic tumor, T1-201 uptake was decreased after radiation therapy. T1-201 brain SPECT could distinguish the benign and malignancy, and seems to be useful in the follow-up after treatment. But one of the early or delayed SPECT seems not to be necessary for these purposes.

• 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.