• Biomolecules & Therapeutics
• /
• 제23권4호
• /
• pp.367-373
• /
• 2015

Cordyceps species including Cordyceps bassiana are a notable anti-cancer dietary supplement. Previously, we identified several compounds with anti-cancer activity from the butanol fraction (Cb-BF) of Cordyceps bassiana. To expand the structural value of Cb-BF-derived anti-cancer drugs, we employed various chemical moieties to produce a novel Cb-BF-derived chemical derivative, KTH-13-amine-monophenyl [4-isopropyl-2-(1-phenylethyl) aniline (KTH-13-AMP)], which we tested for anti-cancer activity. KTH-13-AMP suppressed the proliferation of MDA-MB-231, HeLa, and C6 glioma cells. KTH-13-AMP also dose-dependently induced morphological changes in C6 glioma cells and time-dependently increased the level of early apoptotic cells stained with annexin V-FITC. Furthermore, the levels of the active full-length forms of caspase-3 and caspase-9 were increased. In contrast, the levels of total forms of caspases-3, caspase-8, caspase-9, and Bcl-2 were decreased in KTH-13-AMP treated-cells. We also confirmed that the phosphorylation of STAT3, Src, and PI3K/p85, which is linked to cell survival, was diminished by treatment with KTH-13-AMP. Therefore, these results strongly suggest that this compound can be used to guide the development of an anti-cancer drug or serve as a lead compound in forming another strong anti-proliferative agent.

• Mass Spectrometry Letters
• /
• 제6권3호
• /
• pp.65-70
• /
• 2015

N-Alkyl/benzyl substituted isatin derivatives are intermediates and synthetic precursors for the preparation of biological active heterocycles. N-alkyl/benzyl isatins have showed various biological activities, such as cytotoxicity, antiviral, caspase inhibition, cannabinoid receptor 2 agonists for the treatment of neuropathic pain, etc. In this study, N-alkyl/benzyl isatin derivatives were synthesized from isatin and alkyl/benzyl halides in presence of K₂CO₃ in DMF and excellent to quantitative yields (~95%) were obtained. Isatins and benzyl-isatins were condensed with fluorescein hydrazide to form fluorescein hydrazone. All the compounds were subjected to their fragmentation behavior study using LC/MSⁿ. N-Alkyl substituted isatin derivatives fragmented at nitrogen-carbon (N-C) bond, hence gave daughter ion as [RN+H]⁺. Whereas, N-benzyl substituted isatin derivatives fragmented at carbon-carbon (C-C) bond of alkyl chain which linked with nitrogen molecules, therefore gave N-methyl fragments [RNCH₂]⁺. This study demonstrated that, isatin moiety present in a small/large molecule or in a matrix of reaction mixture with/without N-alkyl/benzyl substituents can be identified by mass spectroscopic fragmentation behavior study.

• Clinical and Experimental Pediatrics
• /
• 제50권7호
• /
• pp.686-693
• /
• 2007

목 적 : Mycophenolate mofetil (MMF)의 활성 대사산물인 (MPA는 IMPDH의 잠재적인 반응 억제제로써 새로운 면역치료제로 사용되고 있다. 이러한 MPA는 신경계에서 흥분독성 손상 후 뇌세포를 보호하고, 미세아교세포에서는 세포사멸사(apoptosis)를 유도하지만, 저산소성 허혈성 뇌질환에서 MPA의 효과는 아직 알려지지 않아, 본 연구에서 Rice-Vannucci 모델을 이용한 신생 백서의 저산소성 허혈성 뇌 손상과 저산소 상태의 태아 백서 뇌세포 배양에서 MPA의 뇌보호 효과를 알아보고자 실험하였다. 방 법 : 생후 7일된 백서의 좌측 총 경동맥을 결찰한 후 저산소 (8% $O_2$) 상태에서 2시간 노출하여, 저산소성 허혈성 뇌 손상을 유발하고 뇌 손상 전후에 MPA(10 mg/kg)를 투여하여 대조군과 비교하였다. 또한, 재태기간 18일된 태아 백서의 대뇌피질 세포를 배양하여 1% $O_2$ 배양기에서 저산소 상태로 세포손상을 유도하여 저산소군, 손상 전후 MPA 투여군($10{\mu}g/mL$)으로 나누어 정상산소군과 비교하였다. 세포사멸사와의 관련을 알아보기 위해서 Bcl-2, Bax, caspase-3 항체로 western blotting하였고 Bcl-2, Bax, caspase-3 primer를 이용하여 real-time PCR을 하였다. 결 과 : 형태학적으로 H&E 염색상 MPA를 투여한 군에서 뇌 보호 효과를 보였다. Western blotting과 real-time PCR을 이용한 저산소성 허혈성 뇌손상 동물 모델뿐만 아니라 저산소 상태로 태아 백서 뇌세포 배양 실험에서도 MPA 투여한 경우 caspase-3의 발현과 Bax/Bcl-2의 비율이 감소함을 보였다. 결 론 : 본 연구에서 MPA가 anti-apoptosis 작용을 통하여 주산기 저산소성 허혈성 뇌 손상에 뇌보호 역할을 하는 것을 알 수 있었고 향후 신생아 저산소성 허혈성 뇌병증의 치료에 임상적 적용이 가능하리라 생각된다.

• Nutrition Research and Practice
• /
• 제12권2호
• /
• pp.93-100
• /
• 2018

BACKGROUND/OBJECTIVE: Oxidative stress plays a key role in neuronal cell damage, which is associated with neurodegenerative disease. The aim of present study was to investigate the neuroprotective effects of perilla oil (PO) and its active component, alpha-linolenic acid (ALA), against hydrogen peroxide $(H_2O_2)$-induced oxidative stress in SH-SY5Y neuronal cells. MATERIALS/METHODS: The SH-SY5Y human neuroblastoma cells exposed to $250{\mu}M$ $H_2O_2$ for 24 h were treated with different concentrations of PO (25, 125, 250 and $500{\mu}g/mL$) and its major fatty acid, ALA (1, 2.5, 5 and $25{\mu}g/mL$). We examined the effects of PO and ALA on $H_2O_2$-induced cell viability, lactate dehydrogenase (LDH) release, and nuclear condensation. Moreover, we determined whether PO and ALA regulated the apoptosis-related protein expressions, such as cleaved-poly ADP ribose polymerase (PARP), cleaved caspase-9 and -3, BCL-2 and BAX. RESULTS: Treatment of $H_2O_2$ resulted in decreased cell viability, increased LDH release, and increase in the nuclei condensation as indicated by Hoechst 33342 staining. However, PO and ALA treatment significantly attenuated the neuronal cell death, indicating that PO and ALA potently blocked the $H_2O_2$-induced neuronal apoptosis. Furthermore, cleaved-PARP, cleaved caspase-9 and -3 activations were significantly decreased in the presence of PO and ALA, and the $H_2O_2$-induced up-regulated BAX/BCL-2 ratio was blocked after treatment with PO and ALA. CONCLUSIONS: PO and its main fatty acid, ALA, exerted the protective activity from neuronal oxidative stress induced by $H_2O_2$. They regulated apoptotic pathway in neuronal cell death by alleviation of BAX/BCL-2 ratio, and down-regulation of cleaved-PARP and cleaved caspase-9 and -3. Although further studies are required to verify the protective mechanisms of PO and ALA from neuronal damage, PO and ALA are the promising agent against oxidative stress-induced apoptotic neuronal cell death.

• Tuberculosis and Respiratory Diseases
• /
• 제57권5호
• /
• pp.449-460
• /
• 2004

연구배경 : PS-341은 최근에 개발된 강력하고 특이적인 proteasome 억제제로서, 일부 암환자에 투여하여 좋은 성적이 보고되고 있다. Proteasome 억제제의 항암효과는 아포프토시스 유발 물질, 즉 p53, $p21^{WAF/CIP1}$, $p27^{KIP1}$, NF-${\kappa}B$, Bax, Bcl-2 등의 발현 증가와 관련이 있는 것으로 생각되고 있다. JNK와 GSK-$3{\beta}$도 아포프토시스에 관여하는 것으로 잘 알려져 있지만, PS-341에 의한 아포프토시스에서의 역할은 규명되지 못한 상태이다. 본 연구에서는 폐암세포주에서 PS-341에 의한 아포프토시스에서 JNK와 GSK-$3{\beta}$의 역할을 규명하고자 하였다. 방 법 : NCI-H157과 A549 폐암세포주를 실험에 사용하였다. 세포생존능은 MTT 방법으로 평가하였고, 아포프토시스는 PARP의 분해로 평가하였다. JNK의 활성도는 in vitro immuno complex kinase 방법과 내인성 c-Jun의 인산화로 측정하였다. 각종 단백의 발현은 Western 분석으로 평가하였다. JNK1과 GSK-$3{\beta}$의 과발현은 각각 plasmid vector와 adenovirus vector를 이용하였다. 결 과 : PS-341 처치로 아포프토시스에 의한 세포생존율의 감소가 관찰되었다. PS-341 처치로 JNK가 활성화되었고, c-Jun의 발현이 유도되었다. Dominant negative JNK1의 과발현 또는 SP600125 전치치로 JNK의 활성화를 차단하면 PS-341에 의한 아포프토시스가 억제되었다. PS-341 처리로 JNK 활성화에 의존적으로 caspase 3의 활성화가 유도되었다. Caspase 활성화의 차단으로도 PS-341에 의한 아포프토시스가 억제되었다. PS-341에 의해 Akt가 활성화되었고, Akt 활성화의 차단으로 PS-341에 의한 아포프토시스가 심화되었다. PS-341에 의한 Akt 활성화로 GSK-$3{\beta}$가 불활성화되었다. Constitutively active GSK-$3{\beta}$의 과발현으로 PS-341에 의한 아포프토시스가 심화되었고, dominant negative GSK-$3{\beta}$의 과발현으로 PS-341에 의한 아포프토시스가 감소되었다. Lithium chloride 전처치와 dominant negative GSK-$3{\beta}$의 과발현으로 PS-341에 의한 JNK의 활성화와 c-Jun의 발현 증가가 억제되었다. 결 론 : 폐암세포주에서 PS-341에 의한 아포프토시스는 JNK/caspase 경로가 관여하며, 이는 PI3K/Akt 경로를 통한 GSK-$3{\beta}$의 불활성화에 의해 억제되는 것으로 판단된다. 따라서 PS-341의 항암효과를 최대화하기 위해서는 PI3K/Akt 경로를 통한 GSK-$3{\beta}$의 불활성화를 차단하는 치료법이 병행되어야 할 것으로 판단된다.

• Journal of Ginseng Research
• /
• 제47권2호
• /
• pp.228-236
• /
• 2023

Background: QiShen YiQi pills (QSYQ) is a Traditional Chinese Medicine (TCM) formula, which has a significant effect on the treatment of patients with myocardial infarction (MI) in clinical practice. However, the molecular mechanism of QSYQ regulation pyroptosis after MI is still not fully known. Hence, this study was designed to reveal the mechanism of the active ingredient in QSYQ. Methods: Integrated approach of network pharmacology and molecular docking, were conducted to screen active components and corresponding common target genes of QSYQ in intervening pyroptosis after MI. Subsequently, STRING and Cytoscape were applied to construct a PPI network, and obtain candidate active compounds. Molecular docking was performed to verify the binding ability of candidate components to pyroptosis proteins and oxygen-glucose deprivation (OGD) induced cardiomyocytes injuries were applied to explore the protective effect and mechanism of the candidate drug. Results: Two drug-likeness compounds were preliminarily selected, and the binding capacity between Ginsenoside Rh2 (Rh2) and key target High Mobility Group Box 1 (HMGB1)was validated in the form of hydrogen bonding. 2 μM Rh2 prevented OGD-induced H9c2 death and reduced IL-18 and IL-1β levels, possibly by decreasing the activation of the NLRP3 inflammasome, inhibiting the expression of p12-caspase1, and attenuating the level of pyroptosis executive protein GSDMD-N. Conclusions: We propose that Rh2 of QSYQ can protect myocardial cells partially by ameliorating pyroptosis, which seems to have a new insight regarding the therapeutic potential for MI.

• Journal of Acupuncture Research
• /
• 제23권2호
• /
• pp.139-157
• /
• 2006

In the present study, I have investigated the bee venom (BV) and melittin (a major component of BV) -mediated anti-proliferative effects, and defined its mechanisms of action in cultured rat aortic vascular smooth muscle cells (VSMCs). BV and melittin $(0.4{\sim}0.8\;{\mu}g/ml)$ effectively inhibited 50 ng/ml platelet derived growth factor BB (PDGF-BB)-induced VSMCs proliferations. The regulation of apoptosis has attracted much attention as a possible means of eliminating excessively proliferating VSMCs. In the present study, the treatment of BV and melittin strongly induced apoptosis of VSMCs. I examined the effects on $NF-{\kappa}B$ activation to investigate a possible mechanism for anti-proliferative effects of BV and melittin, the PDGF-BB-induced $I{\kappa}B{\alpha}$ phosphorylation and its degradation were potently inhibited by melittin, and DNA binding activity and nuclear translocation of $NF-{\kappa}B$ p50 subunit in response to the action of PDGF-BB were potently attenuated by melittin. In further investigations, melittin markedly inhibited the PDGF-BB-induced phosphorylation of Akt but not ERK1/2, upstream signals of $NF-{\kappa}B$. Treatment of melittin also potently induced pro-apoptotic protein p53, Bax, and caspase-3 expression, but decreased anti-apoptotic protein Bcl-2 expression. These results suggest that the anti-proliferative effects of BV and melittin in VSMCs through induction of apoptosis via suppressions of $NF-{\kappa}B$ and Akt activation, and enhancement of apoptotic signal pathway. Based on these results, BV acupuncture can be a candidate as a therapeutic method for restenosis and atherosclerosis.

• 생약학회지
• /
• 제40권1호
• /
• pp.65-69
• /
• 2009

The present study investigated the antiproliferative effect of Litsea japonica in HL-60/ADR, adriamycin resistant human promyelocytic leukemia cells. The 80% ethanol extract of L. japonica markedly inhibited the growth of HL-60/ADR cells. When HL-60/ADR cells were treated with the extract, several apoptosis events like as DNA fragmentation, chromatin condensation and the increase of the population of sub-G1 hypodiploid cells were observed. In the mechanism of apoptosis induction by L. japonica, we examined the changes of Bcl-2 and Bax protein expression levels, and activation of caspases. After the HL-60/ADR cells were treated with the extract, the Bcl-2 expression was decreased, whereas the expression of Bax was increased in a time-dependent manner compared to the control. In addition, the active forms of caspase-9 and -3 were increased and the cleavage of poly (ADP-ribose) polymerase, a vital substrate of effector caspase, was observed. The results suggest that the inhibitory effect of L. japonica on the growth of the HL-60/ADR appears to arise from the induction of apoptosis via the down-regulation of Bcl-2 and the activation of caspases.

• 한국자원식물학회지
• /
• 제31권2호
• /
• pp.102-108
• /
• 2018

본 연구는 꼭두서니의 뿌리인 천초근의 물 추출물과 에탄올 추출물을 이용하여 대장암 세포에 대한 암세포 성장 억제 및 사멸효과가 있는지 알아보고자 수행하였다. ERA(천초근 에탄올 추출물)은 폴리페놀($45.77{\pm}2.03mg/g$)과 플라보노이드($22.82{\pm}1.33mg/g$)를 함유하고 있었으며, $H_2O_2$에 의해 증가된 ROS(reactive oxygen species)를 억제하는 효과를 나타냈지만 WRA(천초근 물 추출물)은 효과가 없었다. 또한 ERA는 $500{\mu}g/m{\ell}$의 농도로 대장암 세포주(HCT-116)에 처리했을 때 세포사멸을 유도할 뿐만 아니라 caspase-3 단백질 활성화, DNA fragmentation 및 apoptotic cell death를 일으키는 것으로 확인되었다. 이는 ERA가 HCT-116 세포주에 대해 apoptosis(세포자멸사)를 통해 항암효과를 나타내는 것으로 생각되지만 다른 연구결과들과 비교하였을 때 농도 대비 효능이 미미하다. 따라서 천초근 에탄올 추출물에 대장암 세포의 성장을 억제하는 유효성분을 분석하여 그 효능을 탐색하는 추가실험이 필요할 것으로 생각된다.

• Molecules and Cells
• /
• 제26권5호
• /
• pp.436-442
• /
• 2008

Osteoclasts are multinucleated cells with the unique ability to resorb bone. Elevated activity of these cells under pathologic conditions leads to the progression of bone erosion that occurs in osteoporosis, periodontal disease, and rheumatoid arthritis. Thus, the regulation of osteoclast apoptosis is important for bone homeostasis. In this study, we examined the effects of the Janus tyrosine kinase 2 specific inhibitor AG490 on osteoclast apoptosis. We found that AG490 greatly inhibited osteoclast apoptosis. AG490 stimulated the phosphorylation of Akt and ERK. Adenovirus-mediated expression of dominant negative (DN)-Akt and DN-Ras in osteoclasts inhibited the survival of osteoclasts despite the presence of AG490. Cytochrome c release during osteoclast apoptosis was inhibited by AG490 treatment, but this effect was inhibited in the presence of LY294002 or U0126. AG490 suppressed the pro-apoptotic proteins Bad and Bim, which was inhibited in osteoclasts infected with DN-Akt and DN-Ras adenovirus. In addition, constitutively active MEK and myristoylated-Akt adenovirus suppressed the cleavage of pro-caspase-9 and -3 and inhibited osteoclast apoptosis induced by etoposide. Taken together, our results suggest that AG490 inhibited cytochrome c release into the cytosol at least partly by inhibiting the pro-apoptotic proteins Bad and Bim, which in turn suppressed caspase-9 and -3 activation, thereby inhibiting osteoclast apoptosis.