• 대한한방내과학회지
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• 제28권2호
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• pp.230-241
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• 2007

Objective : This study was intended to ascertain the protective effect of phenolic-rich fraction (PRF) from Fructus Schisandrae on SH-SY5Y cells. Methods : PRF was obtained from the 80% ethanol extract of Fructus Schisandrae by Sepabeads SP-850 column chromatography. The neuroprotective effect of the FS PRS was investigated due to the hydrogen peroxide $(H_2O_2)-induced$ apoptosis of cultured SH-SY5Y cells. Results : Cell viability assays revealed that pretreating SH-SY5Y cells with PRF (10-200 ${\mu}g/mL$) resulted in significant dose-dependent protection against $H_2O_2-induced$ cell death. The effect was assessed by flow cytometric analysis of DNA contents using propidium iodide (PI) staining. The population of apoptotic cells was increased by 32.89% in only $H_2O_2$ (150 ${\mu}M$)-treated environment, but it was reduced by pre-treatment of FS PRF (200 ${\mu}g/mL$) to 21.61%. $H_2O_2-induced$ caspase-3 activation and PARP cleavage were reduced in FS PRF pre-treated cells, and PRF led to an apparent suppressive effect on the oxidative stress induced by reactive oxygen species (ROS). Conculsion : This study showed that Fructus Schisandrae should be useful for the treatment prevention of neurodegenerative diseases associated with elevated ROS levels.

• 동의생리병리학회지
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• 제22권5호
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• pp.1140-1146
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• 2008

One of the major side effects of cisplatin is nephrotoxicity, leading to acute renal failure. Recent study has suggested a role of hydroxyl radicals and p53 in renal cell injury by cisplatin. This study determined the possible involvement of oxidative stress in p53 activation. In rat mesangial cells, cisplatin treatment induced apoptosis and p53 activation. Pifithrin-$\alpha$, a pharmacological inhibitor of p53, suppressed cisplatin-induced apoptosis. Cisplatin also induced reactive oxidative species (ROS) generation. Of interest, cisplatin-induced apoptosis was prevented by N-acetyl-cysteine (NAC), a general antioxidant. NAC diminished p53 activation during cisplatin treatment. Puerariae Radix and Rehmanniae Radix Preparata with antioxidative activity were reduced the cisplatin-induced ROS generation, caspase-3 activity and p53 activation. In conclusion, ROS may contribute to p53 activation to initiate cisplatin-induced apoptosis in rat mesangial cells. In result, antioxidative effect of Puerariae Radix and Rehmanniae Radix Preparata prevented cisplatin-induced apoptosis through inhibition of p53 activation.

• Toxicological Research
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• 제22권4호
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• pp.315-322
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• 2006

TALP-32 is highly basic protein with a molecular weight of 32 kDa purified from human term placenta. Some basic proteins such as defensins and cecropins are known to induce cell death by increasing membrane permeability and some of them are under development as an anticancer drug especially targeting multi-drug resistant cancers. Therefore, we investigated cytotoxic effect and mechanism of TALP-32 When HeLa cell was incubated with TALP-32, cytotoxicity was increased in time and dose dependent manner. As time goes by, HeLa cells became round and plasma membrane was ruptured. Increase of plasma membrane permeability was determined with LDH release assay. Also in transmission electron microscopy, typical morphology of necrotic cell death, such as cell swelling and intracellular organelle disruption was observed, but DNA fragmentation and caspase activation was not. And necrotic cell death was determined with Annexin V/Pl staining. The cytotoxicity of TALP-32 was minimal and decreased or RBC and Hep3B respectively. These data suggests that TALP-32 induces necrosis on rapidly growing cells but not on slowly growing cells implicating the possibility of its development of anticancer peptide drug.

• 생약학회지
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• 제49권1호
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• pp.47-54
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• 2018

Astilbe rubra (AR) is a perennial, belongs to the Saxifragaceae family, it contains tannin and triterpene. AR has been used in republic korea to improve toxication, fever, pain and convulsion. Recently, number of natural products have been analyzed for potential pharmacological activities including anti-cancer, anti-obesity and anti-diabetic medication. Consequently, we investigated the growth inhibition effect of Astilbe rubra water extract (WAC), ethanol extract (EAC) and bergenin on Hela cell (human adenocarcinoma cell). From whole plant of A. rubra, bergenin was isolated by column chromatography and its structures were identified by $^1H$, $^{13}C$ NMR and IT TOF-ESI MS. High extraction efficiency of bergenin was shown at 0.95% under 60 min reflux extraction with 50% MeOH. The MTS assay showed that EAC (ethanol extract) treatment increased cell death in a dose-dependent manner. Moreover, EAC treatment on Hela cell increased apoptotic cell death and caspase-3 activity. Results suggest that EAC has growth inhibition effect on Hela cells, but not WAC and bergenin. $500{\mu}g/mL$ EAC treatment inhibited Hela cell at $60.2{\pm}1.5%$.

• Journal of Applied Biological Chemistry
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• 제63권2호
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• pp.137-145
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• 2020

Oxidative stress is one of the pathogenic mechanisms of various neurodegenerative diseases, such as Alzheimer's disease. Neuroglia, the most abundant cells in the brain, is thought to play an important role in the antioxidant defense system and neuronal metabolic support against neurotoxicity and oxidative stress. We investigated the protective effect of paeoniflorin (PF) against oxidative stress in C6 glial cells. Exposure of C6 glial cells to hydrogen peroxide (H₂O₂, 500 μM) significantly decreased cell viability and increased amounts of lactate dehydrogenase (LDH) release, indicating H₂O₂-induced cellular damage. However, treatment with PF significantly attenuated H₂O₂-induced cell death as shown by increased cell survival and decreased LDH release. The H₂O₂-stimulated reactive oxygen species production was also suppressed, and it may be associated with improvement of superoxide dismutase activity by treatment with PF. In addition, an increase in ratio of Bcl-2/Bax protein expression was observed after treatment with PF. In particular, the down-stream of the apoptotic signaling pathway was inhibited in the presence of PF, mostly by reduction of cleaved-poly ADP ribose polymerase, cleaved caspase-3, and -9 protein expression. Furthermore, H₂O₂-induced phosphorylation of c-Jun N-terminal kinase and extracellular signal-regulated kinase 1/2 was attenuated by treatment with PF. Taken together, neuroprotective effect of PF against oxidative stress probably result from the regulation of apoptotic pathway in C6 glial cells. In conclusion, our findings suggest that PF may be a potent therapeutic agent for neurodegenerative disorders.

• 동의생리병리학회지
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• 제24권2호
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• pp.220-227
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• 2010

Puerariae radix (PR) is a popular natural herb and a traditional food in Asia, which has antithrombotic and anti-allergic properties and stimulates estrogenic activity. One of the major side effects of cisplatin is nephrotoxicity, leading to acute renal failure. Recent study has suggested a role of ROS and p53 in renal cell injury by cisplatin. We studied that protective effects of PR on cisplatin-induced apoptosis in rat mesangial cell. Rat mesangial cell was preincubated with PR (50, 100, 150 and 200 ${\mu}g/m{\ell}$) for 12 hr and then treated with 30 ${\mu}M$ cisplatin for 24 hr. Protective effect of PR on cisplatin-induced apoptosis in ECV304 cells was determined using MTT assay, FDA-PI staining, flow cytometric analysis, caspase-3 activity assay, ROS assay and western blot. Our results showed that PR inhibited in cisplatin-induced apoptosis and ROS production in ECV304 cells. Moreover, PR reduced ERK, p38 and JNK activation that increased in cisplatin-treated rat mesangial cell. Furthermore, activation of p53 by cisplatin in rat mesangial cell was inhibited by PR treatment. These results suggest that protective effect of PR on cisplatin-induced apoptosis in rat mesangial cell may be associated with reduction of ERK, p38, JNK, p53 activation.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2002년도 창립10주년기념 및 국립독성연구원 의약품동등성평가부서 신설기념 국재학술대회:생물학적 동등성과 의약품 개발 전략을 위한 국제심포지움
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• pp.89-96
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• 2002

The importance of apoptosis in normal development and pathogenesis has been well recognized, and explosive progress towards dissecting its commitment step has been made during the past decade. Mitochondria, Apaf-1, caspase, and bcl-2 family members play central roles in the commitment step. However, it is still unclear how upstream cell survival pathways regulate apoptosis. It is also unknown whether the bcl-2 family members have any effect on the upstream survival pathways. We have demonstrated that the anti-apoptotic gene product bcl-2 greatly induces expression of the tissue inhibitor of metalloproteinase-1 (TIMP-1) in human breast epithelial cells. Surprisingly, we found that TIMP-1, like bcl-2, is a potent inhibitor of apoptosis induced by a variety of stimuli. Functional studies indicate that TIMP-1 inhibits a classical apoptotic pathway mediated by caspases, and that focal adhesion kinase (FAK)/Pl 3-kinase and mitogen activated protein kinase (MAPK) are critical for TIMP- 1 -mediated cell survival. We also showed specific association of TIMP-1 with the cell surface. Consistently, a 150-H)a surface protein was identified in MCF10A cells that specifically binds TIMP-1. Taken together, we hypothesize that TIMP-I binding on the cell surface induces a cell survival pathway that regulates the common apoptosis commitment step. The results of these studies will address a new paradigm in the regulation of apoptosis by an extracellular molecule TIMP-1, and also greatly enhance our understanding of TIMP-1's pleiotropic activity in many physiological and pathological processes. This information may also be useful in designing more rational therapeutic interventions aimed at modulating the anti-apoptotic activity of TIMP-1 .

• Biomolecules & Therapeutics
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• 제23권5호
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• pp.407-413
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• 2015

Paraquat dichloride (N,N-dimethyl-4-4'-bipiridinium, PQ) is an extremely toxic chemical that is widely used in herbicides. PQ generates reactive oxygen species (ROS) and causes multiple organ failure. In particular, PQ has been reported to be an immunotoxic agrochemical compound. PQ was shown to decrease the number of macrophages in rats and suppress monocyte phagocytic activity in mice. However, the effect of PQ on macrophage cell viability remains unclear. In this study, we evaluated the cytotoxic effect of PQ on the mouse macrophage cell line, RAW264.7 and its possible mechanism of action. RAW264.7 cells were treated with PQ (0, 75, and $150{\mu}M$), and cellular apoptosis, mitochondrial membrane potential (MMP), and intracellular ROS levels were determined. Morphological changes to the cell nucleus and cellular apoptosis were also evaluated by DAPI and Annexin V staining, respectively. In this study, PQ induced apoptotic cell death by dose-dependently decreasing MMP. Additionally, PQ increased the cleaved form of caspase-3, an apoptotic marker. In conclusion, PQ induces apoptosis in RAW264.7 cells through a ROS-mediated mitochondrial pathway. Thus, our study improves our knowledge of PQ-induced toxicity, and may give us a greater understanding of how PQ affects the immune system.

• Biomolecules & Therapeutics
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• 제26권2호
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• pp.175-181
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• 2018

Carbon monoxide (CO) is well-known as toxic gas and intrinsic signaling molecule such as neurotransmitter and blood vessel relaxant. Recently, it has been reported that low concentration of CO exerts therapeutic actions under various pathological conditions including liver failure, heart failure, gastric cancer, and cardiac arrest. However, little has been known about the effect of CO in neurodegenerative diseases like Parkinson's disease (PD). To test whether CO could exert a beneficial action during oxidative cell death in PD, we examined the effects of CO on 6-hydroxydopamine (6-OHDA)-induced cell death in C6 glioma cells. Treatment of CO-releasing molecule-2 (CORM-2) significantly attenuated 6-OHDA-induced apoptotic cell death in a dose-dependent manner. CORM-2 treatment decreased Bax/Bcl2 ratio and caspase-3 activity, which had been increased by 6-OHDA. CORM-2 increased phosphorylation of NF-E2-related factor 2 (Nrf2) which is a transcription factor regulating antioxidant proteins. Subsequently, CORM-2 also increased the expression of heme oxygenase-1 and superoxide dismutases (CuZnSOD and MnSOD), which were antioxidant enzymes regulated by Nrf2. These results suggest that CO released by CORM-2 treatment may have protective effects against oxidative cell death in PD through the potentiation of cellular adaptive survival responses via activation of Nrf2 and upregulation of heme oxygenase-1, leading to increasing antioxidant defense capacity.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2011년도 제40회 동계학술대회 초록집
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• pp.265-267
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• 2011

저온 대기압 플라즈마는 21세기에 들어 생의학 분야에 이용될 수 있는 새로운 도구로서 많은 관심을 받고 있다. 대기압 플라즈마는 고가의 진공 장비를 필요로 하지 않고 저전력 구동이 가능하기 때문에 저비용 구동이 가능하고 방전 장치와 전력공급 장치의 소형화에 매우 유리하다. 특히 저온 대기압 플라즈마는 고온의 전자와 저온의 이온 입자가 공존하는 열적 불평형(thermal non-equilibrium) 상태에 있기 때문에 플라즈마의 저온 특성은 유지하면서도 (${\sim}30^{\circ}C$) 물리/화학적 반응성은 매우 높아 그 응용 분야가 매우 넓다. 플라즈마의 다양한 생의학 분야 응용 가운데 세포의 사멸 유발 또는 생장 촉진, 살균/멸균, 지혈, 상처 치유 등에 저온 대기압 플라즈마가 매우 뛰어난 효능을 보인다는 것이 국내외의 다양한 연구를 통해 밝혀지고 있다 [1]. 20 kHz 정현파로 구동되는 플라즈마 장치를 이용한 암 세포 제거 실험에서 플라즈마 처리 효과를 증대시키기 위해 항체-금나노입자 중합체를 암 세포에 주입시켰다 (그림 1(a)). 그 결과 세포의 사멸율은 74%로서 플라즈마 또는 플라즈마-금나노입자만을 처리한 경우에 비하여 사멸율이 매우 높게 나타났다 (그림 1(b)). 이를 통해 암세포 선택성을 가진 항체-금나노입자 중합체와 플라즈마 처리 기술을 융합한 암 세포의 선택적 사멸 유발 기술의 개발 가능성이 열렸다. 또한 플라즈마 처리를 통해 일어나는 세포의 자멸사 기작이 Cytochrome C의 방출 이후 이어지는 Caspase-3의 활성화 경로와 관계가 있음이 밝혀졌다 (그림 1(c)). 치아 미백은 최근 부상하고 있는 저온 대기압 플라즈마의 새로운 응용 분야이다 [5-6]. 대기압에서 동작하는 헬륨 플라즈마 제트를 미백제(과산화수소)와 함께 발치된 치아에 적용하였을 때 (그림 2(a)) 미백제만을 사용하였을 경우에 비해 치아의 색상 변화가 2배 이상 크게 나타나는 것을 확인하였다 (그림 2(b)). 이처럼 최근 그 범위가 크게 넓어지고 있는 저온 대기압 플라즈마의 생의학 응용 기술의 최적화를 위해서는, 다양한 생의학 응용 분야에 따라 요구되는 플라즈마의 특성 및 응용별 기저 기 작에 대한 이해와 연구가 필요하다.