• 생명과학회지
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• 제22권6호
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• pp.760-771
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• 2012

해양 트리테르펜 글리코시드(marine triterpene glycosides)는 해삼(Holothurians)으로부터 분리된 천연물질로서 항 진균작용, 항암작용 및 용혈 작용 등 여러 가지 생물학적 활성들을 가지고 있다고 보고되었다. 또한 이전의 연구 결과 Thelenota anax로부터 분리한 stichoposide C (STC)는 산성 스핑고마이엘리나제와 중성 스핑고마이엘리나제의 활성화에 의한 세라마이드의 생성을 통하여 백혈병 세포주에서 세포사멸을 유도한다는 것을 알 수 있었다. 본 연구에서는 STC와 구조 유사체인 STD가 백혈병 세포에서 세포사멸을 유도하는지와 이에 대한 분자적 기전을 살펴보았다. STC와 STD는 K562 세포와 HL-60 세포에서 농도와 시간 의존적으로 세포 사멸을 일으키고 이러한 세포 사멸은 caspase-8의 활성화, 미토콘드리아 손상, caspase-9의 활성화, 그리고 caspase-3의 활성화에 의해 유도된다. 이러한 결과는 STC와 STD가 외인성 경로와 내인성 경로의 활성화를 통해 세포 사멸을 유도함을 시사한다. 그리고 STC는 산성 SMase와 중성 SMase를 활성화시키고 이 결과로 세라마이드를 생성시킨다. 산성과 중성 SMase의 특이적인 저해제를 이용하여 STC에 의한 세포 사멸이 부분적으로 억제됨을 알 수 있었다. 반면에, STD는 세라마이드 합성 효소의 활성화에 의해서 세라마이드를 생성시킨다. 세라마이드 합성 효소 저해제를 이용하여 STD에 의한 세포 사멸이 부분적으로 억제되는 것을 확인하였다. 더욱이 STC와 STD는 HL-60 세포의 이종 이식 종양 모델에서 종양의 성장을 현저하게 억제하였고 세라마이드의 생성도 증가시켰다. 이러한 결과는 STC와 STD가 aglycone에 부착된 당이 다르므로 서로 다른 경로를 통해 세포 사멸과 항암 활성을 유도한다는 것을 암시하였다. 따라서 이러한 결과는 이들의 작용은 aglycone에 부착된 당에 의해 영향을 받을 수 있고 이들은 향후 백혈병의 치료제로 사용될 수 있다는 것을 제시하였다.

• 대한의생명과학회지
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• 제15권1호
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• pp.61-66
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• 2009

Leflunomide is an immunomodulatory agent used for the treatment of rheumatoid arthritis (RA). Leflunomide known as a regulator of iNOS synthesis which largely decreases NO production in diverse cell type. However, the effect of leflunomide on chondrocyte is still poorly understood. In our previous studies, we have shown that direct production of Nitric oxide (NO) by treating chondrocytes with NO donor, sodium nitroprusside (SNP), causes apoptosis via p38 mitogen-activated protein kinase in association with elevation of p53 protein level, caspase-3 activation. In this study, we characterized the molecular mechanism by which A77 1726 inhibit apoptosis. We found that A77 1726 inhibit NO-induced apoptosis as determined by MTT (Thiazolyl Blue Tetrazolium Bromide) assay and DNA fragmentation. The inhibition of apoptosis by A77 1726 was accompanied by increased PI-3 kinase and AKT activities. So, inhibition of phosphatidylinositol (PI)-3kinase with LY294002 rescued apoptosis. Triciribine, the specific inhibitor of AKT, also abolished anti-apoptotic effect. Our results indicate that A77 1726, the active metabolite of leflunomide, mediates NO-induced apoptosis in chondrocytes by modulating up-regulation of PI-3 kinase and AKT.

• Journal of Ginseng Research
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• 제47권1호
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• pp.23-32
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• 2023

Coronavirus disease 2019 (COVID-19) is a highly infectious respiratory disease caused by a severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2). SARS-CoV-2 infection may cause clinical manifestations of multiple organ damage, including various neurological syndromes. There are currently two oral antiviral drugs-Paxlovid and molnupiravir-that are recognized to treat COVID-19, but there are still no drugs that can specifically fight the challenges of SARS-CoV-2 variants. Nucleotide-binding oligomerization domain-like receptor pyrin domain-containing-3 (NLRP3) inflammasome is a multimolecular complex that can sense heterogeneous pathogen-associated molecular patterns associated with neurological disorders. The NLRP3 activation stimulates the production of caspase-1-mediated interleukin (IL)-1β, IL-18, and other cytokines in immune cells. Panax (P.) ginseng is a medicinal plant that has traditionally been widely used to boost immunity and treat various pathological conditions in the nervous system due to its safety and anti-inflammatory/oxidant/viral activities. Several recent reports have indicated that P. ginseng and its active ingredients may regulate NLRP3 inflammasome activation in the nervous system. Therefore, this review article discusses the current knowledge regarding the pathogenesis of neurological disorders related to COVID-19 and NLRP3 inflammasome activation and the possibility of using P. ginseng in a strategy targeting this pathway to treat neurological disorders.

• BMB Reports
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• 제53권8호
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• pp.419-424
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• 2020

Bee venom (BV), secreted from the venom gland of the honey bee, contains several biological active compounds. BV has been widely used as a traditional medicine for treating human disease, including cancer. In this study, we have shown the molecular mechanism underlying the therapeutic effect of BV on cancer. Treatment with BV reduced the proliferation of cervical-cancer cells in a dose- and time-dependent manner. Interestingly, the killing effect of BV was specific to HPV-positive cervical-cancer cell lines, such as Caski and HeLa cells, and not to HPV-negative cervical-cancer cells (C33A). BV reduced the expression of HPV E6 and E7 at RNA and protein levels, leading to an increase in the expression of p53 and Rb in Caski and HeLa cells. Further, BV decreased the levels of cell-cycle proteins, such as cyclin A and B, and increased the levels of cell-cycle inhibitors, such as p21 and p27. BV significantly induced apoptosis and inhibited wound healing and migration of cervical-cancer cells. It also upregulated the expression of pro-apoptotic BAX and downregulated the expression of anti-apoptotic Bcl-2 and Bcl-XL. Cleavage of caspase-3, caspase-9, and PARP were also induced by BV treatment, whereas the phosphorylation of mitogenic signaling-related proteins, such as AKT, JNK, p38, and ERK, were downregulated. Our results indicate that BV has a therapeutic selectivity for HPV-positive malignant cells, so further clinical studies are needed to assess its clinical application.

• Molecules and Cells
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• 제38권5호
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• pp.416-425
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• 2015

NF-E2-related factor 2 (Nrf2), a basic leucine zipper transcription factor, has recently received a great deal of attention as an important molecule that enhances antioxidative defenses and induces resistance to chemotherapy or radiotherapy. In this study, we investigated the apoptosis-inducing and Nrf2- upregulating effects of quercetin on malignant mesothelioma (MM) MSTO-211H and H2452 cells. Quercetin treatment inhibited cell growth and led to upregulation of Nrf2 at both the mRNA and protein levels without altering the ubiquitination and extending the half-life of the Nrf2 protein. Following treatment with quercetin, analyses of the nuclear level of Nrf2, Nrf2 antioxidant response element-binding assay, Nrf2 promoter-luc assay, and RT-PCR toward the Nrf2-regulated gene, heme oxygenase-1, demonstrated that the induced Nrf2 is transcriptionally active. Knockdown of Nrf2 expression with siRNA enhanced cytotoxicity due to the induction of apoptosis, as evidenced by an increase in the level of proapoptotic Bax, a decrease in the level of antiapoptotic Bcl-2 with enhanced cleavage of caspase-3 and PARP proteins, the appearance of a sub-$G_0/G_1$ peak in the flow cytometric assay, and increased percentage of apoptotic propensities in the annexin V binding assay. Effective reversal of apoptosis was observed following pretreatment with the pan-caspase inhibitor Z-VAD. Moreover, Nrf2 knockdown exhibited increased sensitivity to the anticancer drug, cisplatin, presumably by potentiating the oxidative stress induced by cisplatin. Collectively, our data demonstrate the importance of Nrf2 in cytoprotection, survival, and drug resistance with implications for the potential significance of targeting Nrf2 as a promising strategy for overcoming resistance to chemotherapeutics in MM.

• 생약학회지
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• 제40권1호
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• pp.65-69
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• 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.

• 생명과학회지
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• 제25권4호
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• pp.441-449
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• 2015

본 연구에서는 인체 폐암세포주인 A549 세포와 간암세포주인 HepG2 세포를 사용하여 Endlicheria anomala 메탄올 추출물(Methanol extract of E. anomala, MEEA)의 항암 활성 및 그 분자적 기전에 관하여 분석하였다. 먼저 MEEA가 인체 폐암세포와 간암세포의 증식에 미치는 영향을 분석한 결과, 암세포의 증식을 억제하는 효과가 탁월하였다. 그 후 MEEA에 의한 세포 증식이 억제되는 원인을 분석하기 위하여 Flow cytometry analysis, AnnexinV & 7-AAD 이중 염색을 수행한 결과, 두 세포에서 모두 농도의존적으로 apoptosis 유발군인 SubG1기의 세포 분포가 증가하였고, early apoptosis에서 late apoptosis로 전환되는 공통적인 현상을 확인할 수 있었다. 또한 apoptosis의 유발로 일어날 수 있는 세포의 형태 변화를 관찰하기 위한 DAPI 염색과 DNA fragmentation을 통하여 MEEA 처리에 의한 A549의 염색질 응축, 사멸체 형성 및 DNA의 끌림 현상을 관찰할 수 있었으며, 이와 관련된 분자적 기전 분석을 위한 Western blot을 추가로 수행하여 caspase, PARP, pro-, anti-apoptotic 단백질의 발현을 관찰하였다. 이상의 결과로 MEEA는 인체 폐암세포와 간암세포에서 p53의 발현 증가와 Bcl-2 family의 변화를 유도하며, caspase-3와 관련된 경로를 통하여 농도의존적으로 apoptosis를 유발시킨다는 것을 증명하였다. 이는 MEEA가 항암 활성을 보유하고 있고, 인체 폐암세포와 간암세포 사멸의 기전 연구를 위한 중요한 자료가 될 수 있음을 시사한다.

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

• 대한한의학회지
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• 제34권3호
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• pp.126-142
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• 2013

Objectives: Previous reports have shown that Bogijetong-Tang (BJT) is effective in peripheral neuropathy induced by taxol and crush injury. In this study, we researched the effects of BJT on diabetic neuropathy induced by STZ in the mouse. Methods: We performed both in vitro and in vivo experiments to verify the effects of BJT on diabetic neuropathy induced by STZ in mice. Changes in axonal recovery were observed with immunofluorescence staining using NF-200, Hoechst33258, $S100{\beta}$, caspase 3 and anti-cdc2. Proliferation and degeneration of Schwann cells were investigated by immunofluorescence staining and western blot analyses. Results: BJT showed considerable effects on neurite outgrowth and axonal regeneration in diabetic neuropathy. BJT contributed to the creation of NF-200, GAP-43, Cdc2, phospho-vimentin, ${\beta}1$, active ${\beta}1$, ${\beta}3$ integrin, phospho-Erk1/2 protein. Conclusions: Through this study, we found that BJT is effective for enhanced axonal regeneration via dynamic regulation of regeneration-associated proteins. Therefore, BJT had a pharmaceutical property enhancing recovery of peripheral nerves induced by diabetic neuropathy and could be a candidate for drug development after more research.

• 생명과학회지
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• 제24권9호
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• pp.1001-1005
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• 2014

Ginsenoside Rg3는 홍삼으로부터 추출한 활성 성분들 중 하나로 한방 의학에선 원기를 회복시키는 약제로 잘 알려져 있는 인체에 유효한 화학 성분이다. Rg3는 지금까지 많은 연구들에 의하여 다양한 암세포로부터 강력한 항암효과를 가진다고 알려져 있다. 그러나 Rg3가 악성 흑색종 세포에서 어떻게 세포사멸을 유도하는지에 대한 작용 기작은 명백하게 밝혀지지 않았다. 따라서, 본 연구에서는 ginsenoside Rg3가 B16F10 흑색종 세포에서 세포 사멸 유도 활성 및 기전에 관한 영향을 조사하였다. 세포 생존력을 MTT assay 법으로 수행한 결과, B16F10 세포에선 농도 의존적으로 세포증식 저해 효과가 나타났고 정상세포인 EA.hy.926 과 NIH3T3 에서는 나타나지 않았다. B1610 세포에 Rg3를 농도 별로 처리 후, TUNEL 염색을 한 결과 세포사멸이 농도 의존적으로 증가 하는 것을 확인 할 수 있었다. Western blot 분석을 실시한 결과, Rg3를 처리한 B16F10 세포에서 p-FAK, Bcl-2, pro-caspase3 단백질들의 발현이 감소 되었고 이와 반대로 Bax, p-p38의 발현은 증가되었다. 따라서, 본 연구에서는 Rg3가 B16F10 흑색종 세포에서 항암제의 agent로써 사용 될 수 있다는 것을 입증하였다.