• Applied Biological Chemistry
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• 제51권1호
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• pp.79-83
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• 2008

본 연구에서는 신경독소일 뿐만 아니라 흥분성 신경전달물질로 잘 알려져 있는 glutamate 세포독성이 산화적 손상과 관련하고 있고, 여기에 방어효과를 보이는 제비꽃 추출물에 관하여 연구하였다. MTT reduction assay를 통하여 glutamate의 세포독성을 확인하였고 ascorbic acid와 같은 대표적인 항산화제를 처리한 후 광학 현미경을 이용한 형태학적 변화를 관찰하였다. N18-RE-105 세포주에 최종 농도 20mM의 glutamate를 처리 하면 40.8% 의 생존율을 보이는데 반하여 ascorbic acid 500 ${\mu}M$ 최종농도로 처리하였을 때 85.3%의 세포 생존율을 확인할 수 있었다. 그리고 신경세포 보호효과를 가지는 제비꽃을 methanol, ethanol, acetone 추출한 뒤 MTT reduction assay를 이용하여 활성을 확인하였으며 그 중 acetone 추출물을 최종농도 50, 100 ${\mu}g/ml$를 처리 시 76.8%, 79.4%로 가장 높은 세포 생존율을 확인할 수 있었다. 이 결과는 N18-RE-105 세포주의 형태학적 변화와 LDH release assey에서도 일치하는 결과를 확인하였다.

• 생약학회지
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• 제43권1호
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• pp.72-78
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• 2012

Alnus japonica Steud (A. japonica) have long been used in the traditional medicine for gastric disorder, hepatitis and fatty liver in Korea. Antiulcer effects of A. japonica hot water extract (AJ ext) were evaluated by in vitro antibacterial activity against H. pylori, by the inhibitory action against the in vitro gastric $H^+/K^+$-ATPase and using rat models of gastric mucosal damage and gastric ulcer induced by HCl-ethanol, indomethacin, and restraint and water-immersion stress. For the determination of antibacterial activity of AJ ext against H. pylori, the activity of urease which released from H. pylori was measured in culture. AJ ext showed weak antibacterial activity against H. pylori with the growth inhibitions of 37% and 61% by adding final concentrations of 500 and $1000{\mu}g/ml$ culture, respectively at 24 h. To observe the inhibitory activity of AJ ext against the $H^+/K^+$-ATPase in hog gastric membrane vesicle, $IC_{50}$ value of AJ ext was $806.3{\mu}g/ml$. Pretreatment of AJ ext (200, 500 mg/kg, p.o.) prevented in a dose-dependent manner the acute gastritis in HCl-ethanol model and the formation of gastric ulcer in indomethacin model and restraint and water-immersion stress model. These results suggest that the AJ ext can be used for prevention and treatment of gastric mucosal damage and ulcers induced by various stress.

• Toxicological Research
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• 제35권1호
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• pp.13-24
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• 2019

Numerous ethnomedicinal uses have been attributed to different parts of Annona senegalensis (ASE), including its uses as food and food additives. The present study investigated toxicological and antioxidant effects of 28 days administration of ethanol extracts of ASE stem bark to Wistar strain albino rats. Acute toxicity test was done to determine lethal dose in Wistar rats while sub-acute toxicity test was conducted on rats divided into four groups (A - control, B - 50 mg/kg, C - 100 mg/kg, D - 150 mg/kg, respectively and treated for 28 days. Oxidative stress markers in liver and kidney as well as hepatic succinate dehydrogenase activity in the mitochondrial and post mitochondrial fractions (PMF) were evaluated. The $LD_{50}$ value of ASE was > 2,000 mg/kg. White blood cell counts gradually increased, but red blood cell counts and haematocrits level decreased significantly (p < 0.05) by about 50%. Liver enzymes in the serum and mitochondrial succinate dehydrogenase activity increased significantly (p < 0.05). Superoxide dismutase and catalase activities also increased in liver mitochondria and PMF while malondialdehyde (MDA) and reduced glutathione levels increased only in the PMF. Furthermore, only MDA levels increased significantly in the kidney after 28 days extract administration. Histopathological examination showed hepatic necrosis and no obvious signs of nephrotoxicity. Anona senegalensis is relatively safe, but prolonged ingestion could induce oxidative stress and impair ATP synthesis through the modulation of the activity of mitochondrial succinate dehydrogenase.

• 한방비만학회지
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• 제19권1호
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• pp.1-11
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• 2019

Objectives: The purpose of the present study was to evaluate the preventive effects of ethanol extract of Polygalae radix (EEPR) against oxidative stress (hydrogen peroxide, $H_2O_2$)-induced DNA damage and apoptosis in Chang liver cells. Methods: Chang liver cells were pretreated with various concentrations of EEPR and then challenged with 0.5 mM $H_2O_2$. The cell viability and apoptosis were assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and flow cytometry analysis, respectively. The levels of reactive oxygen species (ROS), mitochondrial membrane potentials (MMPs) and adenosine tri-phosphate (ATP) contents were measured. Expression levels of Bcl-2 and Bax were also determined using Western blot analysis. Results: The results showed that the decreased survival rate induced by $H_2O_2$ could be attributed to the induction of DNA damage and apoptosis accompanied by the increased production of ROS, which was remarkably protected by EEPR. In addition, the loss of $H_2O_2$-induced MMPs and ATP contents was significantly attenuated in the presence of EEPR. The inhibitory effect of EEPR on $H_2O_2$-induced apoptosis was associated with up-regulation of Bcl-2 and down-regulation of Bax, thus reducing the Bax/Bcl-2 ratio. Conclusions: Our data prove that EEPR protects Chang liver cells against $H_2O_2$-induced DNA damage and apoptosis by scavenging ROS and thus suppressing the mitochondrial-dependent apoptosis pathway.

• 대한본초학회지
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• 제36권5호
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• pp.93-99
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• 2021

Objective : Caryophylli Flos has been used in Korean medicine to relieve vomiting and pains caused by chills that make fluid circulation difficult. This study was designed to investigate the protective effect of ethanol extract of Caryophylli Flos (CF) in hydrogen peroxide (H₂O₂)-induced apoptotic cell death in human keratinocyte HaCaT cells. Methods : CF was prepared by extracting 200 g of Caryophylli Flos in 2 L of ethanol for 48 h. Cell viability was measured by MTT assay, and the protein expression was monitored by Western blot analysis. Apoptosis was determined by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Reactive oxygen species (ROS) was measured using fluorescent dye, and reduced glutathione (GSH) was determined with a colorimetric commercial kit. Results : CF protected HaCaT cells from cell death caused by oxidative stress after H₂O₂ treatment. H₂O₂ amplified generation of ROS and induced depletion of GSH, whereas these changes in ROS and GSH were inhibited by GF treatment. In addition, H₂O₂ resulted in apoptosis as assessed by TUNEL assay and the expression of apoptosis regulator proteins. However, cells treated with CF showed a decrease in TUNEL-positive cells and restored the reduced expression of procaspase-9, -3 and PARP. Conclusion : This study showed cytoprotective effects of CF by anti-apoptotic activity while exerting antioxidative activity in H₂O₂-treated HaCaT cells. These results suggest that CF could be beneficial in skin damage caused by oxidative stress.

• 대한본초학회지
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• 제38권1호
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• pp.21-30
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• 2023

Objective : Gardeniae Fructus (GF) has bitter and cold nature. Thus, it has been traditionally prescribed in processed form roasted with ginger juice for patients with a weak stomach. This study investigated the effects of processed GF in tert-butyl hydroperoxide (tBHP)-treated gastric epithelial cells. Methods : Processed GF was made by applying 40% ginger juice or 10% ethanol for 24 h and then roasting at 150℃ for 5 minutes. Apoptosis was determined by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Mitochondrial membrane potential (MMP) was monitored by flow cytometry using the membrane permeable fluorescent dye Rh123. Protein expression was measured by Western blot analysis. Results : Cell viability was reduced by tBHP and restored by ethanol extract of GF (GFE). In the TUNEL assay, it was found that cell death by tBHP was due to apoptosis, and GFE had an anti-apoptotic effect. Processed GF roasted with ginger juice showed the best anti-apoptotic effect. Processed GF also inhibited MMP loss and restored tBHP-induced changes in expression levels of apoptosis-related proteins. Increased ROS production and GSH depletion after tBHP treatment were significantly reduced by processed GF. In addition, tBHP-induced activation of mitogen-activated protein kinase (MAPK) was inhibited by processed GF. Conclusion : These results demonstrate that the processed GF is able to protect gastric epithelial cells from oxidative stress-induced cell death with antiapoptotic and antioxidant activity. In addition, it shows that the processing of GF, which have been traditionally used for gastrointestinal protection, partially have scientific validity.

• Natural Product Sciences
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• 제19권2호
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• pp.179-185
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• 2013

Polygalae radix (PR) has traditionally been used as a sedative and anti-stress agent in oriental countries for a long time. PR which contains many ingredients is especially rich in saponins. This study was performed to investigate whether ethanol extract of PR enhances pentobarbital-induced sleep behaviors. In addition, possible mechanisms also were investigated. PR inhibited locomotor activity in mice. PR increased sleep rate and sleep time by concomitant administration with sub-hypnotic dose of pentobarbital (28 mg/kg). PR prolonged total sleeping time, and shortened sleep latency induced by pentobarbital (42 mg/kg). In addition, PR increased intracellular chloride concentration in primary cultured neuronal cells. The expression level of glutamic acid decarboxylase (GAD) were increased, and ${\gamma}$-aminobutyric acid $(GABA)_A$ receptors subunits were modulated by PR, especially increasing ${\gamma}$-subunit expression. In conclusion, PR augments penobarbital-induced sleep behaviors through activation of $GABA_A$ receptors and chloride channel complex.

• 대한본초학회지
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• 제21권4호
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• pp.159-170
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• 2006

Objectives : Anti-oxidation activities of Malvae Semen and Abutili Semen were compared and evaluated. Methods : We tested the anti-oxidant effect through in vitro experiment and in vivo experiment that induced oxidative stress using ethanol. Results : 1. In the anti-oxidative activities of Malvae Semen and Abutili Semen they had high effects in case of the scavenging activitie of DPPH and superoxide anion radical, and inhibitory effect of linoleic acid oxidation. 2. Malvae Semen had large quantities of phenolic compound than Abutili Semen, but the anti-oxidative activity of Malvae Semen was weaker than that of Abutili Semen. 3. The anti-oxidative effect of Abutili Semen was more strong than that of Malvae Semen in the rat stressed by ethanol. Conclusion : Anti-oxidation activities of Abutili Semen were stronger than those of Malvae Semen. Therefore, the complementary usage of Malvae Semen and Abutili Semen can be considered.

• 식품기술
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• 제23권2호
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• pp.214-219
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• 2010

Saccharomyces cerevisiae는 전통적으로 알코올 음료와 bioethanol 생산에 이용되지만, 발효가 진행되는 동안 효모의 에탄올 생성은 에탄올의 축적에 의한 충격으로 세포활성에 손상을 초래한다. 본 연구는 S. cerevisiae의 에탄올 스트레스 반응과 에탄올 내성의 분자적 기초에 관해 수행되었으며, 에탄올 스트레스가 진행되는 동안 효모의 에탄올 생성 향상을 위한 유전 공학 전략의 수립에 활용될 수 있다. 이전의 연구들은 유전자 발현에 대한 에탄올 스트레스의 충격이 환경적 영향을 받기 때문에 다양한 균주와 조건들에 관해 이루어졌다. 그러나 에탄올 공격에 의해 영향을 받은 gene ontology 범주에서의 일부 공통점은 S. cerevisiae의 에탄올 스트레스 반응이 해당과정 및 미토콘드리아 기능과 관련된 유전자 발현의 증가와 에너지가 요구되는 성장과정과 관련된 유전자의 발현 감소에 따라 에너지 생산에 제약 받음을 의미한다. Genomewide screens를 이용한 연구는 vacuole function의 유지가 에탄올 내성에 대해 중요함을 암시한다. 아마도 단백질 turnover와 이온 항상성 유지에 이 세포기관의 역할이 중요하기 때문인 것으로 사료된다. 특히 에탄올 스트레스가 일어날 때 핵 내 Asr1과 Rat8의 축적은 비록 이 가설이 논란이 많은 주제로 남아있지만 S. cerevisiae가 에탄올 스트레스에 대한 특별한 반응을 가지고 있음을 의미한다.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2002년도 추계학술대회
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• pp.76-78
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• 2002

Heat shock proteins (HSPs) are induced in most living cells by mild heat treatment, ethanol, heavy metal ions and hypoxia. In yeast Saccharomyces cerevisiae, mild heat pretreatment strongly induces Hsp104 and thus provide acquired thermotolerance. The ability of hsp104 deleted mutant $({\triangle}hsp104)$ to acquire tolerance to extreme temperature is severely impaired. In providing thermotolerance, two ATP binding domains are indispensible, as demonstrated in ClpA and ClpB proteases of E. coli. The mechanisms by which Hsp104 protects cells from severe heat stress are not yet completely elucidated. We have investigated regulation of mitochondrial metabolic pathways controlled by the functional Hsp104 protein using $^{13}C_NMR$ spectroscopy and observed that the turnover rate of TCA cycle was enhanced in the absence of Hsp104. Production of ROS, which are toxic to kill cells radiply via oxidative stress, was also examined by fluorescence assay. Mitochondrial dysfunction was manifested in increased ROS levels and higher sensitivity for oxidative stress in the absence of Hsp104 protein expressed. Finally, we have identified mitochondrial complex I and Ferritin as binding protein(s) of Hsp104 by yeast two hybrid experiment. Based on these observations, we suggest that Hsp104 protein functions as a protector of oxidative stress via either keeping mitochondrial integrity, direct binding to mitochonrial components or regulating metal-catalyzed redox chemistry.