• The Korean Journal of Physiology and Pharmacology
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• 제18권4호
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• pp.297-305
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• 2014

Flavonoids have an ability to suppress various ion channels. We determined whether one of flavonoids, cyanidin-3-glucoside, affects adenosine 5'-triphosphate (ATP)-induced calcium signaling using digital imaging methods for intracellular free $Ca^{2+}$ concentration ([$Ca^{2+}$]i), reactive oxygen species (ROS) and mitochondrial membrane potential in PC12 cells. Treatment with ATP ($100{\mu}M$) for 90 sec induced [$Ca^{2+}$]i increases in PC12 cells. Pretreatment with cyanidin-3-glucoside ($1{\mu}g/ml$ to $100{\mu}g/ml$) for 30 min inhibited the ATP-induced [$Ca^{2+}$]i increases in a concentration-dependent manner ($IC_{50}=15.3{\mu}g/ml$). Pretreatment with cyanidin-3-glucoside ($15{\mu}g/ml$) for 30 min significantly inhibited the ATP-induced [$Ca^{2+}$]i responses following removal of extracellular $Ca^{2+}$ or depletion of intracellular [$Ca^{2+}$]i stores. Cyanidin-3-glucoside also significantly inhibited the relatively specific P2X2 receptor agonist 2-MeSATP-induced [$Ca^{2+}$]i responses. Cyanidin-3-glucoside significantly inhibited the thapsigargin or ATP-induced store-operated calcium entry. Cyanidin-3-glucoside significantly inhibited the ATP-induced [$Ca^{2+}$]i responses in the presence of nimodipine and ${\omega}$-conotoxin. Cyanidin-3-glucoside also significantly inhibited KCl (50 mM)-induced [$Ca^{2+}$]i increases. Cyanidin-3-glucoside significantly inhibited ATP-induced mitochondrial depolarization. The intracellular $Ca^{2+}$ chelator BAPTA-AM or the mitochondrial $Ca^{2+}$ uniporter inhibitor RU360 blocked the ATP-induced mitochondrial depolarization in the presence of cyanidin-3-glucoside. Cyanidin-3-glucoside blocked ATP-induced formation of ROS. BAPTA-AM further decreased the formation of ROS in the presence of cyanidin-3-glucoside. All these results suggest that cyanidin-3-glucoside inhibits ATP-induced calcium signaling in PC12 cells by inhibiting multiple pathways which are the influx of extracellular $Ca^{2+}$ through the nimodipine and ${\omega}$-conotoxin-sensitive and -insensitive pathways and the release of $Ca^{2+}$ from intracellular stores. In addition, cyanidin-3-glucoside inhibits ATP-induced formation of ROS by inhibiting $Ca^{2+}$-induced mitochondrial depolarization.

• 대한약리학회지
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• 제29권1호
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• pp.85-96
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• 1993

화학물질에 의한 간암 유발과정에서 transferrin receptor (TfR)의 변동을 밝히기 위해 간을 부분절제한 정상백서의 재생간과 발암물질로 3-Me-DAB를 8주간 투여한 백서 또는 약물 투여 후 부분 간절제 수술을 행하여 세포분열을 유도시킨 백서 간조직으로부터 parenchymal cell (PC)과 nonparenchymal cell (NPC)를 분리하고 각각의 세포막을 제조하여 $^{125}I-transferrin$ 결합실험을 실시한 바 다음과 같은 성적을 얻었다. 1. 3-Me-DAB 투여에 의하여 간조직에서 oval cell의 증식, 재생성 변화, 결절형성, 담관의 증식 및 담관세포암 등의 현저한 조직학적 변화가 동반되었다. 그러나 간세포증식을 더욱 촉진시키기 위하여 부분간절제 수술을 하였을 때 수술 후 경과에 따른 형태학적 변동은 큰 차이가 없었다. 2. 정상 재생간의 PC 및 NPC homogenate에서 transferrin 결합량은 부분간 절제 수술 후 1일 및 3일에 증가되었으며 수술 후 7일에 정상으로 회복되었다. 3-Me-DAB 투여에 의해 두세포군에서 모두 정상세포보다 높게 나타났으며 재생기간에 따라 계속 증가되었다. 3. 정상간의 NPC 세포막에서 transferrin 최대 결합량 (Bmax)은 PC 세포막에서 보다 많이 분포되어 있었으며, Kd는 양세포막에서 5.05 또는 6.3nM로 비슷하였다. 4. 재생간의 NPC 및 PC 세포막에서 transferrin 결합량은 부분 간절제 수술 후 1일 및 3일에 $40{\sim}50%$ 증가되었고 수술 후 7일에 정상치로 회복되었다. 5. 3-Me-DAB 처치에 의하여 NPC 및 PC 세포막의 transferrin 결합량은 정상 간세포막에서 보다 약 3배 증가되었고, 3-Me-DAB 투여후 재생간의 NPC 세포막에서는 부분 간절제 수술 후 3일까지 증가된 후 감소되는 양상인데 반해 PC 세포막에서는 수술 후 7일까지 계속 증가되었다. 6. 3-Me-DAB 투여 후 NPC 및 PC 세포막 transferrin binding site에서 Kd치가 $3.1{\sim}4.1\;nM$과 $25.4{\sim}54.1\;nM$인 두 종류가 존재하는 것으로 나타났다. 이상의 실험성적으로 TfR는 1) 간조직의 PC 및 NPC 세포에 모두 분포되어 있으며, 2) 정상 재생간 및 3-Me-DAB의 처리 후 간세포에서의 세포막 TfR의 증가는 세포내 합성량의 증가에 의하여 일어나며, 3) 정상 재생간의 세포막 TfR는 한 종류의 high affinity site $(Kd,\;<3.1{\sim}7.5\;nM)$에 의하여 증가되나, 3-Me-DAB 처리 후 간세포막에서는 정상에서와 같은 high affinity형 이외에 affinty가 낮은 다른 형태의 TfR $(Kd,\;25.4{\sim}54.1\;nM)$가 세포막으로 출현됨으로써 크게 증가되는 것으로 사료된다.

• 동의생리병리학회지
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• 제17권1호
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• pp.85-90
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• 2003

We investigated the effects of Oak smoke flavoring (OSF, Holyessing) on the growth of DU145 and PC-3 human prostate carcinoma cells. OSF treatment resulted in a concentration-dependent growth inhibition in both DU145 and PC3 cell lines. The anti-proliferative effect of OSF treatment was associated with the induction of apoptotic cell death which was confirmed by morphological change such as membrane shrinking, rounding up and chromatin condensation in DU145 and PC-3 cells. DNA flow cytometry analysis confirmed that OSF treatment increased population of apoptotic sub-G1 phase. Furthermore, we observed an increase of pro-apoptotic protein Bax expression and a decrease of anti-apoptotic protein Bcl-2 by OSF treatment in a dose-dependent manner. OSF also induced a proteolytic cleavage of specific target proteins such as poly(ADP-ribose) polymerase (PARP) and β-catenin proteins. The present results indicated that OSF-induced inhibition of human prostate carcinoma cell proliferation is associated with the induction of apoptosis.

• 한국환경성돌연변이발암원학회지
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• 제25권1호
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• pp.19-24
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• 2005

The Swedish double mutation (KM670/671NL) of amyloid precursor protein (Swe-APP) is associated with early-onset familial Alzheimer's disease (FAD) and increases amyloid beta peptide production. Although APP/A/3 mediated neurotoxicity is observed both in vitro and in vivo, the relationship between mutant APP expression, A/3 production, and neuronal death observed in the brains of FAD patients remains to be elucidated. In this study, we investigated the mechanisms of Swe-APP-induced cell death in HEK293 and NGF-differentiated PC 12 cells. We found that the expression of Swe-APP induced cytochrome C relase, activation of caspase 3 in HEK 293 and NGF-differentiated PC 12 cells. We also show that the reactive oxygen species (ROS) was detected in Swe-APP expressing HEK 293 cells and NGF-differentiated PC 12 cells and that pretreatment with vitamine E attenuated the cellular death, cytochrome C release induced by Swe-APP expression, indicating the involvement of free radical in these processes. These results suggest one of possible apoptotic mechanisms of Swe-APP which could occur through cytochrome C release from mitochondria and this apoptosis inducing effects could be at least in part, due to ROS generation by Swe-APP expression.

• Journal of Digestive Cancer Research
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• 제1권1호
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• pp.29-35
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• 2013

복막암종증(Peritoneal carcinomatosis)은 복강내에 암세포가 파종되어 벽쪽 복막과 내장쪽 복막 표면에 악성세포가 축적되는 상태로 정의할 수 있으며, 이것은 암성복수와도 관련 있다. 일반적으로 복막암종증은 원발암의 전이성 암과 유사하게 치료하나 같은 원발암의 다른 장기로의 전이암과는 달리 예후가 좋지 않다. 보존적 치료만 했을 경우 복막암종증 환자의 중앙생존기간은 3-6개월이다. 복막암종증은 예후가 좋지 않고 치료 방법이 제한적이어서 일선에서 치료하는 내과 의사에겐 여전히 어려운 과제이기도 하다. 최근에 이와 관련된 치료 방법이 많이 연구되어 육안적 병소제거술 및 복막절제술(cytoreductive surgery with peritonectomy)과 함께 조기 수술 후 복강내 화학요법(early postoperative intraperitoneal chemotherapy, EPIC) 또는 수술 중 복강내 온열화학요법(hyperthermic intraperitoneal chemotherapy, HIPEC)을 시행하여 생존율을 향상시키고 있다. 본 논문에서는 복막암종증의 전반적인 특징, 증상, 예후 및 진단과 수술적 방법, 항암 화학요법 등의 치료법에 대하여 알아보고자 한다.

• The Korean Journal of Physiology and Pharmacology
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• 제17권1호
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• pp.57-64
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• 2013

Cells can resist and even recover from stress induced by acute hypoxia, whereas chronic hypoxia often leads to irreversible damage and eventually death. Although little is known about the response(s) to acute hypoxia in neuronal cells, alterations in ion channel activity could be preferential. This study aimed to elucidate which channel type is involved in the response to acute hypoxia in rat pheochromocytomal (PC12) cells as a neuronal cell model. Using perfusing solution saturated with 95% $N_2$ and 5% $CO_2$, induction of cell hypoxia was confirmed based on increased intracellular $Ca^{2+}$ with diminished oxygen content in the perfusate. During acute hypoxia, one channel type with a conductance of about 30 pS (2.5 pA at -80 mV) was activated within the first 2~3 min following onset of hypoxia and was long-lived for more than 300 ms with high open probability ($P_o$, up to 0.8). This channel was permeable to $Na^+$ ions, but not to $K^+$, $Ca^+$, and $Cl^-$ ions, and was sensitively blocked by amiloride (200 nM). These characteristics and behaviors were quite similar to those of epithelial sodium channel (ENaC). RT-PCR and Western blot analyses confirmed that ENaC channel was endogenously expressed in PC12 cells. Taken together, a 30-pS ENaC-like channel was activated in response to acute hypoxia in PC12 cells. This is the first evidence of an acute hypoxia-activated $Na^+$ channel that can contribute to depolarization of the cell.

• Applied Biological Chemistry
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• 제51권3호
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• pp.228-232
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• 2008

본 연구에서는 항산화 황성을 가지면서 신경세포 보호 효과를 가지고 있는 석곡, 형개, 감초 및 작약 추출물을 대상으로 GST 유도 활성 효과를 실험하였다. 그 결과, 감초 추출물이 가장 높은 GST 유도 활성을 나타내었다. 감초를 methanol, ethanol, acetone으로 분획 추출한 뒤 GST 유도 활성을 살펴본 결과, 감초의 methanol 추출물을 50 ${\mu}g/ml$ 농도로 처리 시 대조구에 비해 2.23배로 가장 논은 GTS 유도환성이 나타났다. 그리고 감초의 각 추출물들은 처리 농도에서 세포의 형태학적인 변화를 유도하지 않았으며, 이는 LDH release assay 결과를 통해서도 매우 낮은 세포 독성을 확인할 수 있었다. 감초의 GST 유도 활성 물질들의 용매특성을 알아보기 위하여 methanol 추출물을 극성도에 따라 용매 분획하여 GST 유도 활성을 확인한 결과, diethyl ether층 분획물에서 가장 높은 GST 유도 활성을 나타냄을 확인하였다.

• Journal of Ginseng Research
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• 제45권4호
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• pp.473-481
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• 2021

Background: Mitochondrial dysfunction is one of the significant reasons for Alzheimer's disease (AD). Ginsenosides, natural molecules extracted from Panax ginseng, have been demonstrated to exert essential neuroprotective functions, which can ascribe to its anti-oxidative effect, enhancing central metabolism and improving mitochondrial function. However, a comprehensive analysis of cellular mitochondrial bioenergetics after ginsenoside treatment under Aβ-oxidative stress is missing. Methods: The antioxidant activities of ginsenoside Rb₁, Rd, Re, Rg₁ were compared by measuring the cell survival and reactive oxygen species (ROS) formation. Next, the protective effects of ginsenosides of mitochondrial bioenergetics were examined by measuring oxygen consumption rate (OCR) in PC12 cells under Aβ-oxidative stress with an extracellular flux analyzer. Meanwhile, mitochondrial membrane potential (MMP) and mitochondrial dynamics were evaluated by confocal laser scanning microscopy. Results: Ginsenoside Rg₁ possessed the strongest anti-oxidative property, and which therefore provided the best protective function to PC12 cells under the Aβ oxidative stress by increasing ATP production to 3 folds, spare capacity to 2 folds, maximal respiration to 2 folds and non-mitochondrial respiration to 1.5 folds, as compared to Aβ cell model. Furthermore, ginsenoside Rg1 enhanced MMP and mitochondrial interconnectivity, and simultaneously reduced mitochondrial circularity. Conclusion: In the present study, these results demonstrated that ginsenoside Rg₁ could be the best natural compound, as compared with other ginsenosides, by modulating the OCR of cultured PC12 cells during oxidative phosphorylation, in regulating MMP and in improving mitochondria dynamics under Aβ-induced oxidative stress.

• Asian-Australasian Journal of Animal Sciences
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• 제5권3호
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• pp.527-532
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• 1992

The present paper describes temporal changes of immunohistochemical localization and quantities of carbonic anhydrase isozyme II (CA-II) prochymosin (PC) and pepsinogen (PN) in goat's abomasal mucosae during long term milk feeding. The CA-II was not detected by day 14 after birth and then became positive on day 34 in the parietal cells, suggesting that the excretion of the hydrochloric acid (HCl) begins between days 14 and 34 under a feeding condition without solid materials. The quantity of the PC in the gastric chief cells detected by the ELISA showed rapid increase from the day of birth, making a peak on day 8 and then gradually decreased with age. The decrease in quantity of PC became started during the time period when HCl excretion had not started yet. The quantities of PN in the gastric chief cells were almost stable during the whole period examined. Expressions of these gastric enzymes did not seem to be regulated by the change of feeding condition.

• 대한의생명과학회지
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• 제23권1호
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• pp.39-43
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• 2017

In this study, we evaluated the effects of various hypothermic conditions ($32^{\circ}C$), including lithium chloride treatment, on insulin-like growth factor 1 (IGF-1) gene expression in PC12 cells. The results show that short-term hypothermic treatment (<1 day) resulted in relatively higher IGF-1 gene expression than did longer-term treatment (>1 day). Repeated switching between normal temperature and hypothermia every 2 h increased IGF-1 gene expression approximately 3-4-fold. These findings indicate that hypothermia dynamically regulates IGF-1 gene expression. This study could be helpful for the development of treatment and diagnostic strategies for ischemia.