• 동의신경정신과학회지
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• 제11권2호
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• pp.1-9
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• 2000

In previous studies, heat shock has been reported to induce the apoptosis or programmed cell death through the activation of caspase-3. 1 investigated the effect of juniper pure essential oil on the heat shock-induced apoptosis in human astrocyte cell line CCF-STTGI. Treatment of the astrocytes with heat shock markedly induced apoptotic cell death. However, pretreatment of the astrocytes with juniper oil ingibited the heat shock-induced apoptosis. To determine whether juniper inhibits the heat shock-induced activation of these apoptotic proteases, activation of CPP32 was assessed by Western blotting. Consistent with flow cytometry. DNA fragmentation and giemsa staining, heat shock-induced activation of CPP32 was blocked by juniper oil. Poly(ADP-ribose) polymerase (PARP), cysteine protease substrates were fragmented as a consequence of apoptosis by heat shock. Juniper oil inhibited the PARP fragmentation. This juniper oil also inhibited the heat shock-induced activation of caspase-3. These results suggest that juniper oil may modulate the apoptosis through the activation of the interleukin-1-converting enzyme-like protease.

• 동의신경정신과학회지
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• 제11권1호
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• pp.37-46
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• 2000

We investigated the effects of lemon pure essential oils on the heat shock-induced apoptosis in human astrocyte cell line CCF-STTG1. In previous studies, hear shock has been reported to induce the apoptosis or programmed cell death through the activation of caspase-3. Treatment of CCF-STTG1 cells with heat shock markedly induced apoptotic cell death as determined by flow cytometry. Interestingly, pretreatment of CCF-STTG1 cells with lemon pure essential oils inhibited the heat shock-induced apoptosis. Lemon also inhibited the heat shock-induced apoptosis in primary cultured rat astrocytes. To determine whether lemon inhibits the heat shock-induced activation of these apoptotic proteases, activation of CPP32 was assessed by Western blotting. Consistent with flow cytometry, DNA fragmentation and giemsa staining, heat shock-induced activation of CPP32 was blocked by lemon pure essential oil. PARP, cysteine protease substrates were fragmented as a consequence of apoptosis by heat shock. Lemon oil inhibited the PARP fragmentation. This essential oil also inhibited the heat shock-induced activation of caspase-3. These results suggest that lemon pure essential oils may modulate the apoptosis through the activation of the ICE-like caspases.

• 대한치과교정학회지
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• 제33권6호
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• pp.453-463
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• 2003

본 연구는 MC3T3El 조골세포가 저산소증에 반응하여 유발될 수 있는 세포 고사조절 기전을 구명하고자 함에 목적이 있다. $2\%$ 저산소증의 조건하에서 MC3T3El 조골세포는 DNA 사다리 분절 헝성을 보였으며 형광성 염료인 Hoechst 33258로 염색된 핵 구조 형태 관찰시 시간이 지남에 따라 세포고사 현상을 관찰할 수 있었다 Pancaspase 억제제인 Z-VAD-FMK나 특정한 caspase-3 억제제인 Z-DEVD-CHO로 사전 처치하였을 경우에는 저산소증에 의한 DNA 사다리 분절형성이 농축에 비례하여 억제되었다. caspase-3류의 프로테아제(DEVDase) 활성 증가가 세포고사 중에 관찰되었으나 caspase-1 (YVADase)의 활성은 없었다. 어떤 caspase가 세포고사에 관여하는지를 확인하기 위하여 anti-caspase-3 또는 anti-caspase-6의 항체를 이용한 western blotting이 시행되었다. caspase-3의 활성산물에 해당하는 17-KDa단백질과 caspase-6의 활성산물인 20-KDa 단백질이 세포용해물에서 발생되었다. 또한 시간 경과와 더불어 caspase-6의 활동의 상징인 Lamin A의 분열을 일으켰으며, 사이토크롬 C를 cytosol로 방출하였다. 이로써 저산소증에 의한 조골세포의 고사 과정에 사이토크롬 C의 방출이 포함된 caspase의 활성이 관여한다는 것을 확인할 수 있었다.

• 동의신경정신과학회지
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• 제10권2호
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• pp.47-57
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• 1999

Recently, essential oils are used for aromatherapy. Most essential oils are said to be anti-bacterial; some may be anti-viral or anti-fungal. I investigated the effects of peppermint pure essential oil on the heat shock-induced apoptosis in human astrocyte cell line CCF-STTGI. In previous studies, heat shock has been reported to induce the apoptosis or programmed cell death through the activation of caspase-3. We studied the heat shock-induced apoptosis through flow cytometry, DNA electrophoresis, and giemsa staining. Interestingly, these events were inhibited by pretreatment of peppermint pure essential oils in CCF-STTGl cells. Peppermint oil also inhibited the heat shock-induced apoptosis in primary cultured rat astrocytes. In addition, this Peppermint essential oil inhibited the heat shock-induced activation of caspase-3. These results suggest that peppermint pure essential oils may modulate the apoptosis through the activation of the interleukin-I -converting enzyme-like protease.

• 대한한방내과학회지
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• 제23권2호
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• pp.181-190
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• 2002

The water extract of Omija-tang(OMJT) has been traditionally used for treatment of ischemic heart and brain damage in oriental medicine. However, little is known about the mechanism by which the water extract of OMJT protects cells from such damage. Therefore, this study was conducted to investigate the protective mechanisms of OMJT on $H_2O_2$-induced toxicity in H9c2 cardiomyoblast cells. Treatment of $H_2O_2$ markedly induced death of H9c2 cardiomyoblast cells in a dose-dependent manner. The characteristics of $H_2O_2$-induced death of H9c2 showed apparent apoptotic features, such as DNA fragmentation. However, OMJT significantly reduced both $H_2O_2$-induced cell death and chromatin fragmentation. The decrease of Bcl-XL expression by $H_2O_2$ was inhibited by OMJT. In addition, the increase of Bcl-XS and Bax expression were also inhibited by OMJT. In particular, Fas expression, which is generally recognized as cell death inducing signal by Fas/FasL interaction, was markedly increased by $H_2O_2$ in a time-dependent manner, whereas this increase was completely prevented by OMJT. The combined treatment of OMJT and $H_2O_2$ in H9c2 cells also reduced activation of caspase-9 and caspase-3 like protease. Taken together, this study indicates that the protective effects of the water extract of OMJT against oxidative damage may be mediated by the modulation of BcI-XL/S and Bax expression by way of the regulation of mitochondrial membrane potential and caspase cascades.

• 동의생리병리학회지
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• 제18권6호
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• pp.1652-1660
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• 2004

Apoptosis of vascular smooth muscle cells(VSMCs) is essential in atherogenesis, being a factor that modulates its early progression rather than a terminal event in the course of the disease. Various stimuli, including oxide lipoproteins, altered hemodynamic stress and free radical, can induced VSMCs apoptosis in vitro. The protective effects of Sophorae Radix (SR) on apoptotic cell death induced by H₂O₂ were investigated in VSMCs. The viability of VSMCs was markedly decreased by H₂O₂. Sophorae Radix protected the H202-induced apoptotic death of VSMCs, which was characterized as nuclear fragmentation and increase of sub-G0/G1 fraction .. Sophorae Radix decreased the activation of caspase-3 like protease induced by H₂O₂ and recovered control level from H202-induced PARP, Bak, Bcl-XL and mitochondrial membrane potential. These results suggest that Sophorae Radix protected VSMCs apoptotic death induced by H₂O₂ via inactivation of caspase-3 and modulation of mitochondrial function. Also, the expression profile of proteins by using two-dimensional (2-D) gel electrophoresis was screened. Future investigations will need to explore the use of an anti atherosclerotic therapy of Sophorae Radix, which relies on inhibition of the proapoptotic activation of the vascular smooth muscle cells.

• 동의생리병리학회지
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• 제18권2호
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• pp.544-552
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• 2004

In oriental medicine, Radix Polygalae(RP) has been to treat tremors et al. But the mechanism how to decrease tremors was not known. The purpose of this study was to investigate the effect of RP on neurodegenerative disease. We used RP to execute the study of this defense mechanism on dopamine-induced cell death in human SH-SY5Y dopaminergic neuroblastoma cells. MTT assay was used to know the cytotoxicity of dopamine and the defense mechanism. As a result of this experiment, dopamine had cytotoxicity in human SH-SY5Y cells, but when it treated with RP, the cell survival rate increased. This suppressed the cell apoptosis, activation of caspase-3 protease, production of ROS, and repair of membrane potential change. In conclusion, RP has the protective effect on dopamine-induced cell death in human SH-SY5Y dopaminergic neuroblastoma cells, so this could be an effective agent on the neurodegenerative disease like Parkinsonism.

• 대한한의학회지
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• 제24권2호
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• pp.179-192
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• 2003

Objectives : Boyanghwanoh-tang (Buyanhaiwu-tang) has been used as a prescription for stroke, senile and vascular dementia, ischemic brain and heart damage in Oriental traditional medicine. However, there is little known about the mechanism by which the water extracts of Boyanghwanoh-tang (Buyanhaiwu-tang) rescue cells fromthese damages, and little is known about the protective mechanisms of Boyanghwanoh-tang (Buyanhaiwu-tang) on oxidative stress in neuronal cells. Therefore, we have investigated the role of Boyanghwanoh-tang (Buyanhaiwu-tang) on serum and glucose deprived apoptosis in PC12 cells. Methods : PC12 Cells have been used extensively as a model for studying the cellular and molecular effects of neuronal cells. The viability of cells was measured by MIT assay. We used DNA fragmentation and caspase 1, 2, 3, 6, 9-likeproteases activation assay. Transcriptional activation of NF-kB was assessed by using electrophoretic mobility shift assay. Results : Boyanghwanoh-tang (Buyanhaiwu-tang) rescued PC12 cells from apoptotic death by serum and glucose deprivation in a dose-dependent manner. The nuclear staining of PC12 cells clearly showed that Boyanghwanoh-tang (Buyanhaiwu-tang) attenuated nuclear condensation and fragmentation, which represent typical neuronal apoptotic characteristics. Boyanghwanoh-tang (Buyanhaiwu-tang) also prevents fragmentation of genomic DNA and activation of caspase 3-like protease in serum and glucose deprived PC12 cells. Furthermore, Boyanghwanoh-tang (Buyanhaiwu-tang) reduced the activation of NF-kB by serum and glucose-deprived apoptosis. Conclusions : These findings suggest that serum and glucose deprivation induces reduced glutathione (GSH) depletion, and consequently, apoptosis through endogenously produced reactive oxygen species in PC12 cells. Also, our data indicated that Boyanghwanoh-tang (Buyanhaiwu-tang) has protective effects against the serum and glucose deprived deaths of PC12 cells, which are mediated by the generation of GSH that, in turn, can reduce oxidative stress caused by reactive oxygen species (ROS) such as hydrogen peroxide.