• Natural Product Sciences
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• 제21권2호
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• pp.134-140
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• 2015

The present study aims to investigate the effect of methanol extract of Korean mistletoe (KM; Viscum album var. coloratum), on amyloid $\beta$ protein ($A\beta$) (25-35), a synthetic 25-35 amyloid peptide, -induced neurotoxicity in cultured rat cerebral cortical neurons and memory impairment in mice. Exposure of cultured neurons to $10{\mu}M$ $A\beta$ (25-35) for 24 h induced a neuronal cell death, which was measured by a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay and Hoechst 33342 staining. KM (10, 30 and $50{\mu}g/ml$) significantly inhibited the $A\beta$ (25-35)-induced apoptotic neuronal death. KM ($50{\mu}g/ml$) inhibited 10 μM Aβ (25-35)-induced elevation of intracellular calcium concentration ([Ca²⁺]_i), which was measured by a fluorescent dye, Fluo-4 AM. Glutamate release into medium and generation of reactive oxygen species (ROS) induced by $10{\mu}M$ $A\beta$ (25-35) were also inhibited by KM (10, 30 and $50{\mu}g/ml$). These results suggest that KM may mitigate the $A\beta$ (25-35)-induced neurotoxicity by interfering with the increase of [Ca²⁺]_i and then inhibiting glutamate release and generation of ROS in cultured neurons. In addition, orally administered KM (25 and 50 mg/kg, 7 days) significantly prevented memory impairment induced by intracerebroventricular injection of $A\beta$ (25-35) (8 nmol). Taken together, it is suggested that anti-dementia effect of KM is due to its neuroprotective effect against $A\beta$ (25-35)-induced neurotoxicity and that KM may have therapeutic role in prevention of the progression of Alzheimer's disease.

• The Korean Journal of Physiology and Pharmacology
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• 제17권4호
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• pp.259-265
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• 2013

The anti-apoptotic effect of (-)-epigallocatechin-3-gallate (EGCG) during unilateral testicular torsion and detorsion (TT/D) was established in our previous study. In mice, the smallest inhibitor of apoptosis, survivin, is alternatively spliced into three variants, each suggested to have a unique function. Here, we assessed how EGCG exerts its protective effect through the expression of the different survivin splice variants and determined its effect on the morphology of the seminiferous tubules during TT/D. Three mouse groups were used: sham, TT/D+vehicle and TT/D treated with EGCG. The expression of the survivin variants (140 and 40) and other apoptosis genes (p53, Bax and Bcl-2) was measured with semi-quantitative RT-PCR. Histological analysis was performed to assess DNA fragmentation, damage to spermatogenesis and morphometric changes in the seminiferous tubules. In the TT/D+vehicle group, survivin 140 expression was markedly decreased, whereas survivin 40 expression was not significantly different. In parallel, there was an increase in the mRNA level of p53 and the Bax to Bcl-2 ratio in support of apoptosis induction. Histological analyses revealed increased DNA fragmentation and increased damage to spermatogenesis associated with decreased seminiferous tubular diameter and decreased germinal epithelial cell thickness in the TT/D+vehicle group. These changes were reversed to almost sham levels upon EGCG treatment. Our data indicate that EGCG protects the testis from TT/D-induced damage by protecting the morphology of the seminiferous tubules and modulating survivin 140 expression.

• Journal of Ginseng Research
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• 제36권3호
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• pp.248-255
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• 2012

Potential antioxidant effect of processed ginseng (sun ginseng, SG) on oxidative stress generated by tert-butyl hydroperoxide (t-BHP) was investigated in HepG2 cells. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and lactate dehydrogenase (LDH) leakage test demonstrated that SG dose-dependently prevents a loss of cell viability against t-BHP-induced oxidative stress. Also, SG treatment dose-dependently relieved the increment of activities of hepatic enzymes, such as aspartate aminotrasferase and alanine aminotransferase, and lipid peroxidation mediated by t-BHP treatment in HepG2 cells. SG increased the gene expression of antioxidant enzymes such as superoxide dismutase, catalase, and glutathione peroxidase. However, high dose of SG treatment caused decrease in mRNA level of glutathione peroxidase as compared to low dosage of SG-treated cells. The gene expression of glutathione reductase was found to be slightly increased by SG treatment. In addition, SG extract attributed its hepaprotective effect by inducing the mRNA level of bcl-2 and bcl-xL but reducing that of bax. But, the gene expression of bad showed no significant change in SG-treated HepG2 cells. These findings suggest that SG has hepatoprotective effect by showing reduction of LDH release, activities of hepatic enzymes and lipid peroxidation and regulating the gene expression of antioxidant enzymes and apoptosis-related molecules against oxdative stress caused by t-BHP in HepG2 cells.

• 한국약용작물학회지
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• 제19권1호
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• pp.31-37
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• 2011

Previous work demonstrated that an ethanol extract (HS0608) of a mixture of three medicinal plants of Curcuma longae radix, Phellinus linteus, and Scutellariae radix markedly inhibits $A{\beta}$ (25-35)-induced neurotoxicity. The present study was performed to further verify the neuroprotective effect of HS0608 on oxidative and ischemic cerebral injury using cultured rat cortical neurons and rats. Exposure of cultured cortical neurons to $100\;{\mu}M$ hydrogen peroxide ($H_2O_2$) induced neuronal apoptotic death. At $10-100{\mu}g/ml$, HS0608 inhibited neuronal death, elevation of intracellular calcium concentration ($[Ca^{2+}]_i$), and generation of reactive oxygen species (ROS) induced by $H_2O_2$ in primary cultures of rat cortical neurons. In vivo, HS0608 prevented cerebral ischemic injury induced by 2-h middle cerebral artery occlusion (MCAO) and 24-h reperfusion. The ischemic infarct and edema were significantly reduced in rats that received HS0608 (200 mg/kg). These results suggest that the anti-oxidative properties of HS0608 may be responsible for its neuroprotective effect against focal cerebral ischemic injury and that HS0608 may have a therapeutic role in neurodegenerative diseases such as stroke.

• 대한한의학회지
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• 제30권4호
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• pp.79-92
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• 2009

Objectives: The aim of the present study was to explore the neuroprotective effect and the possible mechanism of the PD-1 extracts on 1-methyl-4-phenyl-1,2,3,6-tetrahydrophridine (MPTP)-lesioned C57BL/6 mouse model of Parkinson's disease (PD). Methods: The mice were supplemented (or not) with 50 or 100 mg/kg/day of PD-1 for 2 weeks, after which MPTP was injected intraperitoneally. We observed that daily administration of PD-1 prevented MPTP-induced depletion of striatal DA, and maintained striatal and nigral tyrosine hydroxylase (TH) protein levels. Results: Our results demonstrated that mice treated with PD-1 prior to MPTP administration showed more abundant TH-immunopositive (TH-ir) fibers and neurons than mice given only MPTP, indicating that PD-1 protects dopaminergic striatal fibers and nigral neurons from MPTP insults. Possible neuroprotective effect of PD-1 was further studied by the detection of antiapoptotic protein (bcl-2) and proapoptotic protein (Bax). In this assay, MPTP elevated the Bax protein and decreased the bcl-2 protein, while these expressions were prevented by PD-1 pre-treatment. Conclusions: The present results suggest that PD-1 is able to protect dopaminergic neurons from MPTP-induced neuronal injury with anti-apoptotic activity being one of the possible mechanisms.

• 대한본초학회지
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• 제29권3호
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• pp.71-78
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• 2014

Objectives : This study evaluates the neuroprotective effects of Sopung-tang, a mixture of Notopterygii Rhizoma, Saposhnikoviae Radix, Angelicae Gigantis Radix, Cnidii Rhizoma, Hoelen, Aurantii Nobilis Pericarpium, Pinelliae Tuber, Linderae Radix, Angelicae Dahuricae Radix, Cyperi Rhizoma, Cinnamomi Ramulus, Asari Radix, Glycyrrhizae Radix on the cerebral infarct combined with hyperlipidemia. Method : The hyperlipidemia was induced by the beef tallow 30% diet for 14 days on Sprague-Dawley rats. The cerebral infarct was induced by the middle cerebral artery occlusion (MCAO) for 2 hours with intraluminal thread method. Then the water extract of Sopung-tang was administered a day for 5 days at 3 hours after the cerebral infarct by MCAO. Effect of Sopung-tang was evaluated with the infarct volume and edema percentage by a TTC-stained brain section, and the expressions of Bax and Bcl-2 in the brain tissue by a immunohistochemical stain method. Results : Sopung-tang reduced the infarct size partly in a TTC-stained brain section of the hyperlipidemic MCAO rats. Sopung-tang reduced the infarct volume of the hyperlipidemic MCAO rats significantly. Sopung-tang reduced the edema percentage of the hyperlipidemic MCAO rats, but not significant statistically. Sopung-tang suppressed the Bax expressions in the cerebral penumbra and caudate putamen of the hyperlipidemic MCAO rats significantly. Sopung-tang upregulated the Bcl-2 expression in the caudate putamen of the hyperlipidemic MCAO rats. Conclusion : These results suggest that Sopung-tang plays an anti-apoptotic neuroprotective effect through the suppression of Bax and up-regulation of Bacl-2 expressions in the brain tissues.

• BMB Reports
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• 제55권10호
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• pp.506-511
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• 2022

Advanced hepatocellular carcinoma (HCC) is among the most challenging cancers to overcome, and there is a need for better therapeutic strategies. Among the different cancer drugs that have been used in clinics, sorafenib is considered the standard first-line drug for advanced HCC. Here, to identify a chemical compound displaying a synergistic effect with sorafenib in HCC, we screened a focused chemical library and found that MG149, a histone acetyltransferase inhibitor targeting the MYST family, exhibited the most synergistic anticancer effect with sorafenib on HCC cells. The combination of sorafenib and MG149 exerted a synergistic anti-proliferation effect on HCC cells by inducing apoptotic cell death. We revealed that cotreatment with sorafenib and MG149 aggravated endoplasmic reticulum (ER) stress to promote the death of HCC cells rather than adaptive cell survival. In addition, combined treatment with sorafenib and MG149 significantly increased the intracellular levels of unfolded proteins and reactive oxygen species, which upregulated ER stress. Collectively, these results suggest that MG149 has the potential to improve the efficacy of sorafenib in advanced HCC via the upregulation of cytotoxic ER stress.

• 생명과학회지
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• 제31권4호
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• pp.389-399
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• 2021

이 연구는 인간진피섬유아(HDF)세포에서 병풀 및 아시아티코사이드가 H₂O₂ 유래 세포주기 정지기, 미토콘드리아 활성 및 염증성 사이토카인에 미치는 영향을 조사하였다. 병풀 80% 메탄올 추출물, 에틸아세티이트 분획물 및 병풀의 대표물질인 아시아티코사이드를 사용하였다. 병풀 추추물, 에틸아세테이트 분획 및 아시아티코사이드로 처리한 세포는 낮은 수준의 TNF-α 및 IL-6을 분비하였고, 아시아티코사이드의 항산화 효과는 병풀 추출물 및 에틸아세테이트 분획물보다 높았다. 아시아티코사이드 처리는 미토콘드리아의 막포텐셜을 증가시키고, 미토콘드리아를 정상으로 되돌렸다. 스트레스 유도 후 에틸아세테이트 분획물 및 아시아티코사이드에 의해 미토콘드리아 산소 소비율이 증가하였고, TLR4-MyD88-TRAF6-p65 경로가 재감소하였다. 이러한 결과는 병풀 추출물, 에틸 아세테이트 분획 및 아시아티코사이드가 HDF 세포의 미토콘드리아 활성을 조절할 뿐 아니라 항산화 및 항염증에 효과 있음을 시사한다.

• 생명과학회지
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• 제24권11호
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• pp.1244-1251
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• 2014

길경(桔梗, Platycodon grandiflorum)은 도라지의 뿌리로 항염증, 항알러지, 면역 반응, 당뇨, 고지혈증 및 항암 효과 등을 가지고 있는 것으로 알려져 있다. 하지만 길경의 항암 효과에 대한 연구는 미미하며, 길경이 유발하는 autophagy에 대한 연구는 되어 있지 않다. 본 연구에서는 HCT-116 대장암 세포주에서 길경 추출물이 autophagy와 apoptosis를 유발하면서 세포 성장을 억제하는지의 여부를 조사하였다. 길경 추출물은 농도 및 시간의존적으로 세포의 증식을 억제하였으며, 길경 추출물에 의해 나타나는 apoptosis는 caspase의 활성이 부분적으로 관여되어 있음을 알 수 있었다. 또한, 길경 추출물의 처리는 autophagy에 의해 나타나는 공포를 형성하면서 autophagy와 관련되어 있는 여러 단백질의 발현 조절 및 LC3 단백질의 축적이 동반되었다. 길경 추출물에 의해 유도되는 autophay와 apoptosis의 관계를 알아보기 위해서 3-MA나 bafilomycin A1을 처리하여 autophagy를 억제하였을 때 apoptosis가 유의적으로 증가됨을 알 수 있었다. 흥미롭게도 bafilomycin A1을 처리한 결과에서 길경 추출물에 의한 세포성장 억제가 뚜렷하게 회복되는 양상을 보였다. 따라서 본 연구의 결과는 HCT-116 세포에서 길경 추출물에 의해 유도된 autophagy는 세포 보호적인 작용이 아닌 autophagic cell death이며, 길경 추출물이 대장암 세포주에서 암세포의 사멸을 유도하는 효과적인 대안이 될 수 있음을 알 수 있었다.

• 생명과학회지
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• 제28권1호
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• pp.37-42
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• 2018

NAG-1 단백질은 TGF-${\beta}$ superfamily 유전자로서 암세포의 apoptosis를 유도하고 항암 활성과 관련이 있는 것으로 알려져 있다. 본 연구에서는 프로폴리스 유래의 파이토케미칼 CAPE (caffeic acid phenethyl ester)에 의한 항암유전자 NAG-1의 발현과 발현조절에 대해 연구하였다. 인간 대장암 세포주 HCT116에서 CAPE의 처리에 의해 농도의존적, 시간의존적으로 NAG-1의 발현이 증가됨을 확인하였다. 게다가, 다른 대장암 세포주인 LOVO 세포주에서도 농도의존적으로 NAG-1의 발현이 증가됨을 확인하였다. p53-null HCT116세포주를 이용한 실험에서 CAPE에 의한 NAG-1의 발현은 전사조절인자인 p53에 의존하지 않음을 증명하였다. 또한, 3가지 종류의 NAG-1 프로모터 construct를 이용한 실험에서, cis-element 후보가 -474와 -1,086사이에 있음을 증명하였다. CAPE에 의해 전사조절인자인 ATF3와 CREB의 발현이 변화되는 지를 확인한 결과, CREB은 전혀 발현이 증가되지 않는 반면 ATF3는 CAPE 처리에 의해 농도의존적으로 발현이 증가함을 확인하였다. 그리고, pCG-ATF3와 pCREB의 cotransfection 실험에서 CREB은 NAG-1의 발현에 영향을 못 미치는 반면, ATF3의 과대발현에 의해 NAG-1의 발현이 증가됨을 확인하였다. 결론적으로, CAPE에 의한 NAG-1의 발현은 주로 전사조절인자인 ATF3를 경유하여 일어남을 시사한다.