• 대한한방내과학회지
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• 제30권1호
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• pp.51-63
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• 2009

Background and Objective : The prospects for developing an anti-apoptotic natural component or a compound that exerts a neuroprotective effect with few or no side effects for the treatment of neurodegenerative disease appear favorable. In the present study, we evaluated the effects of the methanol extract of Phellodendri Cortex (PC extract) on 1-methyl-4-phenyl pyridinium($MPP^+$)-induced apoptosis in PC-12 cells. Materials and Methods : We used the methanol extract of Phellodendri Cortex (PC extract). PC-12 cells were cultured by RPMZ-1640. We found the PC extract's gene expression (Bax, Bcl-2) by using RT-PCR. We examined the PC extract's protein expression (Bcl-2, Bax, cytochrome c, poly (ADP-ribose) polymerase (PARP), caspase-3) by SDS-PAGE and Western blot. Results : Apoptosis in $MPP^+$-induced PC-12 cells was accompanied by an increased Bax/Bcl-2 ratio, release of cytochrome c to the cytosol and the activation of caspase-3. PC extract inhibited the down-regulation of Bcl-2 and the up-regulation of Bax, as well as the release of mitochondrial cytochrome c into the cytosol. In addition, PC extract attenuated caspase-3 activation and cleavage of poly (ADP-ribose) polymerase (PARP). Conclusion : These results suggest that the neuroprotective potentials of PC extract against $MPP^+$-induced apoptosis can be. at least partially, ascribed to its anti-apoptotic effects in PC-12 cells.

• 대한본초학회지
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• 제28권2호
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• pp.33-38
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• 2013

Objects : Apoptosis leads to the death of a cell. The mitochondrial pathway of apoptosis, a process regulated by the Bcl-2 family of proteins, plays a key role in various biological processes. The tuber of Liriope platyphylla Wang et TANG (Liliaceae), also known as Liriopis tuber, is famous in Oriental medicine owing to its tonic, antitussive, expectorant and anti-asthmatic properties. The purpose of this study was to observe the effect of the Liriopis tuber on mitochondrial-mediated apoptosis in PC-12 cells. Method : A cytotoxic test on Liriopis Tuber water extract was conducted and another MTT assay was conducted to observe the cytoprotective effect against 4-HNE 25 ${\mu}M$ that causes oxidative stress in PC-12 cells for 24 hours. In addition, in order to observe the expression of Bcl-2 and Bax protein involved with apoptosis, western blot was conducted. Results : The LT water extract had no toxicity for PC-12 cell. In the cytoprotective effect against 4-HNE, both of the group treated with 50 ${\mu}g/m{\ell}$ and 100 ${\mu}g/m{\ell}$ of LT water extract showed a significant increase in comparison with the control group. In Bax protein expression, all the experimental groups treated with LT water extract showed a decrease in comparison with the control group but had no significance. In Bcl-2 protein expression, all the experimental groups treated with LT water extract showed a significant increase in comparison with the control group. Conclusion : These results suggest that LT is effective in reducing apoptosis.

• Radiation Oncology Journal
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• 제20권3호
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• pp.246-252
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• 2002

목적 : 전리 방사선에 의해 유도되는 세포의 apoptosis에서 Fas:Fas ligand 상호 작용의 역할이 보고되고 있다. 본 연구는 Fas 변이를 보이는 Ipr 마우스를 이용하여 in vivo에서 Fas의 발현이 전리 방사선에 의해 유도되는 apoptosis에서 어떤 역할을 하는지 조사하고자 하였다. 대상 및 방법 : Fas의 변이를 보이는 $MRL/Mpj-Fas^{Ipr}$ 마우스와 이의 대조군으로서 MRL/Mpj 마우스를 대상으로 하였다. 마우스는 8주령 웅성으로서 이들에게 전신 방사선을 조사하고 일정 시간 후 비장을 적출하였다. 조직을 hematoxylin-eosin 염색하여 apoptosis 유도 수준을 비교 분석하였다. 또한 apoptosis의 조절 물질인 p53, Bcl-2, Bax, $Bcl-X_L,\;Bcl-X_S$에 대하여 Western blotting을 시행하고 발현수준을 densitometry로 분석하여 관련된 기전을 연구하였다. 결과 : 25 Gy 방사선 조사 후 MRL/Mpj 마우스는 8시간째에 apoptosis가 최대로 많이 일어났고 24시간째에 거의 정상 수준에 가깝게 회복이 되었다. 반면 $MRL/Mpj-Fas^{Ipr}$ 마우스에서는 4시간째에 apoptosis가 최대로 많이 일어났고 8시간째부터 회복되기 시작하였다. MRL/Mpj 마우스의 경우 apoptosis의 최대 유도 수준은 25 Gy 조사 후 8시간째에 $52.3{\pm}7.8%$이었으나, $MRL/Mpj-Fas^{Ipr}$ 마우스는 같은 시간대에 $8.0{\pm}8.6\%$로서 $MRL/Mpj-Fas^{Ipr}$ 마우스에서 apoptosis의 유도 수준이 유의하게 대조군 보다 낮은 것으로 나타났다(p<0.05). 유전물질의 발현에서 25 Gy 방사선 조사후 MRL/Mpj 마우스에서는 p53은 1시간부터 증가를 보여 8시간째에 최대치인 3배를 보였으며, $Bcl-A_L$과 $Bcl-X_S$은 1시간부터 증가를 보여 12시간째에 각각 최대치인 3.3배, 3배를 보였다. 그러나 Bcl-2와 Bax는 뚜렷한 증감을 보이지 않았다. MRL/Mpj 마우스에서 p53, $Bcl-X_L$ 및 $Bcl-X_S$ 등의 발현이 방사선에 의하여 유의하게 증가한 데 반하여 $MRL/Mpj-Fas^{Ipr}$ 마우스에서는 p53, Bcl-2, $Bcl-X_L,\;Bcl-X_S$ 및 Bax 등 분석한 유전자 산물 어느 것도 뚜렷한 증감을 보이지 않았다. 결론 : Fas의 변이가 있는 Ipr 마우스에서 방사선에 의한 apoptosis가 대조군보다 현저하게 낮게 나타났고 이는 방사선에 의한 p53의 유도가 미약한 것과 연관된 것으로 나타났다. 방사선에 의한 apoptosis 유도에 Fas의 역할이 매우 중요한 것으로 보인다.

• Radiation Oncology Journal
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• 제15권3호
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• pp.187-195
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• 1997

목적 : 세포 독성 인자가 유도하는 아포프토시스에 관한 연구가 대부분 In Vitro 연구에 국한되어온 바, In VIVO에서 방사선에 의한 아포프토시스의 유도와 이에 관여하는 유전자들의 발현 양상을 분석하기 위하여 본 연구를 수행하였다. 대상 및 방법 : 마우스 동종암으로서 방사선 민감 종양인 난소암 (OC3-1)과 내성 종양인 간암 (HCa-1)을 모델로 하여 이들 종양이 평균 직경 8 mm로 자랐을 때 25 Gy의 방사선을 조사하였다. 조사 후 다양한 시간 간격으로 조직을 채취하여 아포프토시스의 유도 수준을 분석하며 동시에 이에 관련된 유전자 산물인 p53, $p21^{wart/cip1}$, bax, bel-2 등의 발현을 western blotting 을 이용하여 분석하였다. 종양의 p53 상태는 polymerase chain reaction-single strand conformation polymorphism assay로 분석하였다. 결과 : 모델 종양들의 p53 상태는 둘다 자연형으로 나타났다. 방사선 조사로 OCa-1에서는 아포프토시스가 유도되었으나 HCa-1에서는 아포프토시스가 관찰되지 않았다. OCa-1에서 방사선 조사로 p53, $p21^{wart/cip1}$의 발현이 증가되었으며 bel-2/ bax 비율은 감소하였다. HCa기에서는 p53, $p21^{wart/cip1}$의 발현이 증가되었으나 $p21^{wart/cip1}$은 OCa-1과 비교하여 증가 수준이 미약하였다. bel-2/bax 비율은 현저히 증가하였다. 이와 같은 변화들은 방사선 조사에 선행되거나 조사 후 수시간 내에 일어났으며 아포프토시스의 유도에 선행하거나 일치하였다. 결론 : 아포프토시스의 진행에는 p53, $p21^{wart/cip1}$의 증가 뿐만 아니라 bel-21 bax 비율의 변화 가 관여된다는 것이 In vivo에서 확인되었다. p53가 자연형인 경우에도 그 이하 단계의 유전자 발현 양상이 다르게 나타날 수 있으며 이는 세포 독성 요인을 이용한 암 치료시 결과를 예측하는데 있어서 단일 유전자 발현의 평가와 연계되는 복잡성을 시사하고 있다.

• 대한약침학회지
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• 제18권1호
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• pp.19-26
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• 2015

Objectives: Quercetin, a flavonoid compound, has been reported to induce apoptosis in cancer cells, but its anti-inflammatory effects, which are also closely linked with apoptosis, if any, on non-small-cell lung cancer (NSCLC) have not so far been critically examined. In this study, we tried to determine if quercetin had any demonstrable anti-inflammatory potential, which also could significantly contribute to inducing apoptosis in a NSCLC cell line, A549. Methods: In this context, several assays, including cytotoxicity, flow cytometry and fluorimetry, were done. Gene expression was analyzed by using a western blot analysis. Results: Results revealed that quercetin could induce apoptosis in A549 cells through mitochondrial depolarization by causing an imbalance in B-cell lymphoma 2/Bcl2 Antagonist X (Bcl2/Bax) ratio and by down-regulating the interleukine-6/signal transducer and activator of transcription 3 (IL-6/STAT3) signaling pathway. An analysis of the data revealed that quercetin could block nuclear factor kappa-light-chain-enhancer of activated B cells (NF-${\kappa}B$) activity at early hours, which might cause a down-regulation of the IL-6 titer, and the IL-6 expression, in turn, could inhibit p-STAT3 expression. Down-regulation of both the STAT3 and the NF-${\kappa}B$ expressions might, therefore, cause down-regulation of Bcl2 activity because both are major upstream effectors of Bcl2. Alteration in Bcl2 responses might result in an imbalance in the Bcl2/Bax ratio, which could ultimately bring about mitochondria mediated apoptosis in A549 cells. Conclusion: Overall, the finding of this study indicates that a quercetin induced anti-inflammatory pathway in A549 cells appeared to make a significant contribution towards induction of apoptosis in NSCLC and, thus, may have a therapeutic use such as a strong apoptosis inducer in cancer cells.

• 대한본초학회지
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• 제30권1호
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• pp.59-65
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• 2015

Objectives : The purpose of this study was to observe the effects of Polygalae Radix(PR) on 4-HNE-induced apoptosis in PC-12 cell. Methods : A MTT assay was conducted to observe the cytotoxicity of Polygalae Radix on the cell viability and the cytoprotective effect of Polygalae Radix against 4-HNE that causes oxidative stress-induced cytotoxicity, and then a western blot was conducted to observe the expression of $TNF-{\alpha}$, caspase-3, Bax and Bcl-2 protein that are important factors involved with apoptosis signaling pathway. Results : The Polygalae Radix water extract $25{\mu}g$, $50{\mu}g$, $100{\mu}g$ and $200{\mu}g/mL$ had no cytotoxicity on the PC-12 cell. The Polygalae Radix water extract $25{\mu}g$, $50{\mu}g$ and $100{\mu}g/mL$ had the cytoprotective effect against 4-HNE that causes cytotoxicity on the PC-12 cell. The Polygalae Radix water extract $50{\mu}g/mL$ significantly suppressed the increase in $TNF-{\alpha}$ protein expression in PC-12 cell. The Polygalae Radix water extract $25{\mu}g$ and $50{\mu}g/mL$ significantly suppressed the increase in caspase-3 protein expression in PC-12 cell. The Polygalae Radix water extract $25{\mu}g$, $50{\mu}g$ and $100{\mu}g/mL$ suppressed the increase in Bax protein expression in PC-12 cell but had no significance. The Polygalae Radix water extract $25{\mu}g$ and $100{\mu}g/mL$ significantly prevented the decrease in Bcl-2 protein expression in PC-12 cell, Conclusions : These results suggest that the Polygalae Radix water extract is effective in inhibiting apoptosis.

• 대한한의학방제학회지
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• 제26권3호
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• pp.195-205
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• 2018

Objectives : The purpose of this study was to observe the effects of Gardeniae Fructus on 4-HNE-induced apoptosis in PC-12 cell. Methods : A MTT assay was conducted to observe the cytotoxicity of Gardeniae Fructus on the PC-12 cell viability and the cytoprotective effects of Gardeniae Fructus on PC-12 cell against oxidative stress caused by 4-HNE. And western blot was conducted to observe the expression of Bax, Bcl-2, Caspase-3, $TNF-{\alpha}$ proteins which are involved in intrinsic and extrinsic apoptosis pathway. Results : 25, 50, 100, 200 and $400{\mu}g/m{\ell}$ of Gardeniae Fructus water extract had no cytotoxicity on PC-12 cell. $200{\mu}g/m{\ell}$ of Gardeniae Fructus water extract had significant cytoprotective effect on PC-12 cell against oxidative stress caused by 4-HNE. The expression of Bax protein in 50, 100 and $200{\mu}g/m{\ell}$ of Gardeniae Fructus was significantly decreased in PC-12 cell. The expression of Bcl-2 protein in $200{\mu}g/m{\ell}$ of Gardeniae Fructus was significantly increased in PC-12 cell. The expression of Caspase-3 protein in 100 and $200{\mu}g/m{\ell}$ of Gardeniae Fructus was significantly decreased in PC-12 cell. The expression of $TNF-{\alpha}$ protein in $50{\mu}g/m{\ell}$ of Gardeniae Fructus was significantly decreased in PC-12 cell. Conclusions : These results suggest that Gardeniae Fructus water extract is effective to protect PC-12 cell from 4-HNE induced apoptosis.

• 대한본초학회지
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• 제31권1호
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• pp.77-82
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• 2016

Objectives : The study was designed to observe the effect of Polygalae Radix Preparata Cum Glycyrrhizae Radix on 4-Hydroxynonenal (4-HNE)-induced apoptosis in PC-12 cell.Methods : A cytotoxic test on Polygalae Radix Preparata Cum Glycyrrhizae Radix (PG) was conducted and another MTT assay was conducted to observe the cytoprotective effect against 4-HNE that cause oxidative stress. In addition, in order to observe the expression of Bax, Bcl-2, Caspase-3 and TNF-α protein involved with apoptosis, western blot was conducted.Results : The groups treated with 25 ㎍, 50 ㎍ and 100 ㎍ of PG water extract had no toxicity for PC-12 cell. The groups treated with 25 ㎍, 50 ㎍ and 100 ㎍ of PG water extract showed a significant increase of cell survival rate in comparison with the control group injected by only 4-HNE. The groups treated with 25 ㎍ and 50 ㎍ of PG water extract showed a significant supression on increase of Bax protein expression in the control group. The group treated with 100 ㎍ of PG water extract showed a significant promotion on decrease of Bcl-2 protein expression in the control group. The group treated with 50 ㎍ of PG water extract showed a significant supression on increase of Caspase-3 protein expression in the control group. The group treated with 25 ㎍ of PG water extract showed a significant supression on increase of TNF-α protein expression in the control group.Conclusions : These results suggest that Polygalae Radix Preparata Cum Glycyrrhizae Radix is effective in reducing apoptosis by 4-HNE-dameged cell.

• Advances in nano research
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• 제4권2호
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• pp.145-156
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• 2016

Dendrimers are one of the most appropriate nanocaries for imaging moieties in imaging applications.The purpose of this study was the evalution of cytotoxicity and inducing apoptosis of dendrimers. This study was conducted in order to investigate the metastasis suppression effect of dendrimer in human breast MCF-7 cell line and finding the nanoparticle protein corona in biological enviromental. Dendrimer cytotoxicity effect was assessed by MTT assay. The mRNA experession level of KAI1 as a metastasis suppressor gene, Bax as Pro- apoptotic gene, Bcl-2 as an anti-apoptotic gene and GAPDH as a housekepping gene were determined by real-time PCR assays.concentration-dependent nanoparticle cytotoxicity effect was proofed at range of 1-2 mg/mL in 24 hours, significant upregulation of mRNA expression of Bax, was observed whereas expression of anti-apoptotic Bcl-2 was down-regulated, also expression of metastasis suppressor gene KAI1 was up-regulated. So far a few studies confirmed apoptosis enhancement effect of dendrimers in MCF-7 cell line via bax/bcl-2 pathways. dendrimer nanoparticles was able to act as metastase inhibitor via upregulation of KAI1 gene.

• 한국미생물·생명공학회지
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• 제49권3호
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• pp.283-288
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• 2021

본 연구에서는 굴피나무 잎 추출물로부터 사람 위암세포주인 AGS 세포에 대한 apoptosis가 유도되어 항암 활성을 가지는지를 확인하고자 한다. AGS 세포에 굴피나무 잎 추출물을 농도별(0, 50, 150, 200 ㎍/ml)로 처리하여 세포에서 나타나는 양상들을 확인하였다. MTS 측정으로 암세포 생존율을 확인한 결과 굴피나무 잎 추출물에 따라 농도의존적으로 세포에 독성을 보였으며, 이러한 세포 죽음이 apoptosis 유도에 의한 것인지를 annexin V/FITC-PI 염색을 통해 확인하였다. 그 결과, early apoptosis를 보이는 세포의 양상이 유의성 있게 증가하였다. 굴피나무 잎 추출물의 처리는 세포 주기에서 sub-G1기의 증가로 암세포가 더 이상 분열하지 못하고 증식이 억제됨을 보여준다. 내인성 경로에 관여하는 Bax, Bak, Bcl-2, Bcl-xL의 증감을 mRNA 수준에서 RT-PCR로 확인한 결과, Bax, Bak가 증가하고, Bcl-2, Bcl-xL이 감소를 동반하며 미토콘드리아를 통한 세포사멸의 신호전달 경로가 진행되고 있음을 확인할 수 있었다. 이러한 결과를 종합하면 굴피나무 잎 추출물은 사람 위암세포에 대한 항암 활성을 가지는 효능적 가치가 있음에 가능성을 제시할 수 있을 것으로 생각되며, 추가적인 동물실험을 통한 인체적용시험의 효능 검증을 통해 의약품 개발 가능성을 확인해야 한다.