• 대한한방내과학회지
• /
• 제26권1호
• /
• pp.182-198
• /
• 2005

Object : This study was designed to research whether the protection and inhibitory effects of cardiovascular diseases in L-NAME induced rat or ECV 304 cell lines through the Cell morphological pattern, Tunel assay, LDH activity, heart rate, blood pressure and immunohistochemistric analysis by Boonsimgieum water extract Methods : Nitric oxide(NO) play an important role in normal and pathophysiological cells including as a messenger molecule, neurotransmitter, microbiocidal agent, or dilator of blood vessels and artheriosclerosis, hypertension, myocardial infarction, respectively. Endothelial cell products can modulate the magnitude of a response to a vasoconstrictor, as evinced by the greater constriction after endothelium removal or NO synthesis blockade. To investigate that Boonsimgieum in the potential contribution of the levels of nitric oxide generated by endothelial nitric oxide synthase (eNOS) and the mechanisms of protection against NG-nitro-L-arginine methyl ester (L-NAME), human ECV 304 cells, which normally do not express eNOS, were expressed by L-NAME. L-NAME stimulated rat or cells were found to be resistant to injury and delayed death following the Boonsimgieum. Inhibition of nitric oxide synthesis abolished the protective effect against L-NAME, thrombin and collagen exposure. Interestingly, such effects have been observed during stimulation with agents such as phenylephrine and KCl on L-NAME mediate rats, were damaged by the NOS inhibitor L-NAME. Result : As the result of this study, In group, the anti-apoptosis and necrosis in the cardiovascular system have a potential capacity for prevented, protected and treating the diseases of cardiovascular system, against the necrosis of rat and ECV 304 cells with Caspase 3 and calpain expression by L-NAME is promoted. Conclusion : these results demonstrate neuroprotective and memory enhancing effects of ZIBU, suggesting its beneficial actions for the treatment of AD.

• Journal of Animal Science and Technology
• /
• 제50권4호
• /
• pp.445-456
• /
• 2008

텔로미어란 염색체 말단부에 TTAGGG의 반복 염기서열과 특정 단백질로 구성되어 있는 것으로 핵 내 염색체의 안정성에 작용을 하며 세포의 노화, 사멸 및 암의 발생과 관련이 있다. 텔로머레이스는 텔로미어의 길이를 일정하게 유지하기 위한 직접적 효소로서 telomeric DNA 합성에 관여하는 ribonucleoprotein이다. 본 연구에서는 한우와 Holstein종 136두를 대상으로 백혈구 세포를 이용하여 연령 별, 품종 별, 성 별 텔로미어 함량을 분석하였다. 또한 동일 연령에서 혈액, 간, 뇌, 심장, 신장 및 생식선 조직들의 텔로미어 함량과 텔로머레이스 활성도도 비교 분석하였다. Telomeric DNA의 양적분석은 양적형광접합보인법(Q-FISH)을 이용하였고, 텔로머레이스 활성도의 분석은 TRAP방법을 이용하였다. 분석 결과, 소의 백혈구 세포들에 있어 개체의 연령이 증가함에 따라 텔로미어의 함유율이 점진적이며 유의적으로 감소되는 양상을 보였고, 한우가 Holstein에 비해 텔로미어 함유율이 높게 나타나 품종 간 유의적 차이가 있었으며, 성 간에도 수컷이 암컷에 비해 유의적으로 높은 텔로미어 함유율을 나타내었다. 반면 동일 연령의 간, 심장, 신장, 폐, 혈액 세포내 텔로미어의 함유율은 차이가 없는 것으로 나타났다. 텔로머레이스 활성도는 태아의 모든 조직에서 비교적 강한 활성을 보였지만, 성 성숙이 된 18개월령에서는 생식선 조직을 제외한 나머지 조직에서 텔로머레이스 활성도는 현저하게 떨어져 조직별 세포의 증식성 특이성과 텔로머레이스 활성도간에는 밀접한 연관성이 있는 것으로 나타났다. 이상의 결과로서 세포내 텔로미어 양적 분포 양상 및 텔로머레이스 활성도를 이용하여 개체의 연령 지표 또는 생리적 표지의 개발 가능성을 제시하고 자 한다.

• 생명과학회지
• /
• 제24권7호
• /
• pp.777-783
• /
• 2014

항암제로 알려진 cisplatin과 t-BHP를 토끼에 투여하여 유도된 급성 신부전 시 산조인 추출액을 처리하였을 때 신장 세포의 보호에 미치는 항산화 효과를 조사하였다. 신장을 분리한 후 신피질 절편 실험에서 세포의 손상을 유발하는 지질과산화 및 LDH 실험에서 t-BHP 단독 처리 시 대조군에 비하여 각각 3배, 5배 이상 증가하였으나 산조인 추출액 0.5%를 동시 처리하였을 때는 대조군 수준으로 감소하였다. Creatinine 측정과 지질과산화 실험에서 cisplatin $5mg{\cdot}kg^{-1}$을 복강 투여한 군의 creatinine 농도가 $2.13{\pm}0.1mg{\cdot}dl^{-1}$로 나타났으나 산조인 추출액 $50mg{\cdot}kg^{-1}{\cdot}day^{-1}$을 7일간 전처리 후 cisplatin 투여 48시간 경과한 군은 $0.84{\pm}0.1mg{\cdot}dl^{-1}$로 creatinine의 농도가 약 60% 감소되는 신장보호 효과를 나타내었고, 지질과산화 검사는 cisplatin 단독 투여 시 대조군에 비하여 1.6배 높게 나타났으나 산조인 추출액 전처리 시 1.1배로 대조군과 유사하였다. 병리조직 검사는 cisplatin 단독 처리군에서 근위곱슬세관이 대조군에 대하여 더 붉게 염색되었으며 근위곱슬세관은 내강의 융모세포가 탈락하여 공포를 형성하였다. 그러나 산조인 추출액을 7일간 전처리한 군에서는 근위곱슬세관이 대조군과 유사한 염색소견을 보였고 근위곱습세관도 내강의 융모세포 탈락이 거의 나타나지 않았다. 따라서 cisplatin과 t-BHP에 의해 유발된 신장세포 손상에 대하여 산조인 추출액이 항산화 효과를 보였다.

• Journal of Ginseng Research
• /
• 제40권4호
• /
• pp.423-430
• /
• 2016

Background: The induction of cellular defensive genes such as phase II detoxifying and antioxidant enzymes is a highly effective strategy for protection against carcinogenesis as well as slowing cancer development. Transcription factor Nrf2 (nuclear factor E2-related factor 2) is responsible for activation of phase II enzymes induced by natural chemopreventive compounds. Methods: Red ginseng oil (RGO) was extracted using a supercritical $CO_2$ extraction system and chemical profile of RGO was investigated by GC/MS. Effects of RGO on regulation of the Nrf2/antioxidant response element (ARE) pathway were determined by ARE-luciferase assay, western blotting, and confocal microscopy. Results: The predominant components of RGO were 9,12-octadecadienoic acid (31.48%), bicyclo[10.1.0] tridec-1-ene (22.54%), and 22,23-dihydrostigmasterol (16.90%). RGO treatment significantly increased nuclear translocation of Nrf2 as well as ARE reporter gene activity, leading to upregulation of heme oxygenase-1 and NAD(P)H:quinone oxidoreductase 1. Phosphorylation of the upstream kinases such as apoptosis signal-regulating kinase (ASK)1, mitogen-activated protein kinase (MAPK) kinase (MKK)4/7, c-Jun N-terminal kinase (JNK), and p38 MAPK were enhanced by treatment with RGO. In addition, RGO-mediated Nrf2 expression and nuclear translocation was attenuated by JNK inhibitor SP600125 and p38 MAPK inhibitor SB202190. Conclusion: RGO could be used as a potential chemopreventive agent, possibly by induction of Nrf2/ARE-mediated phase II enzymes via ASK1-MKK4/7-JNK and p38 MAPK signaling pathways.

• Journal of Ginseng Research
• /
• 제44권4호
• /
• pp.664-671
• /
• 2020

Background: Ginsenoside compound-Mc1 (Mc1) is a member of the deglycosylated ginsenosides obtained from ginseng extract. Although several ginsenosides have a cardioprotective effect, this has not been demonstrated in ginsenoside Mc1. Methods: We treated H9c2 cells with hydrogen peroxide (H₂O₂) and ginsenoside Mc1 to evaluate the antioxidant effects of Mc1. The levels of antioxidant molecules, catalase, and superoxide dismutase 2 (SOD2) were measured, and cell viability was determined using the Bcl2-associated X protein (Bax):B-cell lymphoma-extra large ratio, a cytotoxicity assay, and flow cytometry. We generated mice with high-fat diet (HFD)-induced obesity using ginsenoside Mc1 and assessed their heart tissues to evaluate the antioxidant effect and the fibrosis-reducing capability of ginsenoside Mc1. Results: Ginsenoside Mc1 significantly increased the level of phosphorylated AMP-activated protein kinase (AMPK) in the H9c2 cells. The expression levels of catalase and SOD2 increased significantly after treatment with ginsenoside Mc1, resulting in a decrease in the production of H₂O₂-mediated reactive oxygen species. Treatment with ginsenoside Mc1 also significantly reduced the H₂O₂-mediated elevation of the Bax:Bcl2 ratio and the number of DNA-damaged cells, which was significantly attenuated by treatment with an AMPK inhibitor. Consistent with the in vitro data, ginsenoside Mc1 upregulated the levels of catalase and SOD2 and decreased the Bax:B-cell lymphoma-extra large ratio and caspase-3 activity in the heart tissues of HFD-induced obese mice, resulting in reduced collagen deposition. Conclusion: Ginsenoside Mc1 decreases oxidative stress and increases cell viability in H9c2 cells and the heart tissue isolated from HFD-fed mice via an AMPK-dependent mechanism, suggesting its potential as a novel therapeutic agent for oxidative stress-related cardiac diseases.

• Journal of Korean Neurosurgical Society
• /
• 제47권1호
• /
• pp.30-35
• /
• 2010

Objective: The purpose of this study is to explain the effect and reciprocal action among tumor necrosis factor (TNF) like weak inducer of apoptosis (TWEAK), fibroblast growth factor-inducible 14 (Fn14), and transforming growth factor-$\beta1$ (TGF-$\beta1$) on degeneration of human intervertebral disc (IVD). Methods: Human intervertebral disc tissues and cells were cultured with Dulbecco's Modified Eagle's Medium/Nutrient F-12 Ham (DMEM/F-12) media in $37^{\circ}C$, 5% $CO_2$ incubator. When IVD tissues were cultured with TWEAK, Fn14 that is an antagonistic receptor for TWEAK and TGF-$\beta1$, the level of sulfated glycosaminoglycan (sGAG) was estimated by dimethyl methyleneblue (DMMB) assay and sex determining region Y (SRY)-box 9 (Sox9) and versican messenger ribonucleic acid (mRNA) levels were estimated by reverse transcriptase polymerase chain reaction (RT-PCR). Results: When human IVD tissue was cultured for nine days, the sGAG content was elevated in proportion to culture duration. The sGAG was decreased significantly by TWEAK 100 ng/mL, however, Fn14 500 ng/mL did not change the sGAG production of IVD tissue. The Fn14 increased versican and Sox9 mRNA levels decreased with TWEAK in IVD tissue TGF-$\beta1$ 20 ng/mL elevated the sGAG concentration 40% more than control. The sGAG amount decreased with TWEAK was increased with Fn14 or TGF-$\beta1$ but the result was insignificant statistically. TGF-$\beta1$ increased the Sox9 mRNA expression to 180% compared to control group in IVD tissue. Sox9 and versican mRNA levels decreased by TWEAK were increased with TGF-$\beta1$ in primary cultured IVD cells, however, Fn14 did not show increasing effect on Sox9 and versican. Conclusion: This study suggests that TWEAK would act a role in intervertebral disc degeneration through decreasing sGAG and the mRNA level of versican and Sox9.

• 대한한방부인과학회지
• /
• 제19권1호
• /
• pp.14-30
• /
• 2006

Purpose : This study was conducted to investigate the inhibitory effects of Cortex ulmi pumilae on cell proliferation in HeLa cell. Methods : Human uterine cervical carcinoma HeLa cells were cultured in the 1%, 5% and 10% concentration of Cortex ulmi pumilae solution for 24 hours, 48 hours and 72 hours for the direct inhibitory effects of Cortex ulmi pumilae. Afterwards, we executed the analysis of the effect of Cortex ulmi pumilae solution on cell proliferation inhibition using XTT assay, DNA fragmentation, molecular biological method through MAP kinase activity and FACS analysis of caspase activity in the HeLa cells. Results : After 48 and 72 hours cultivation, the HeLa cells showed the concentration-dependently significant increase in all Cortex ulmi pumilae solution containing groups compared to the control. In the FACS analysis, all Cortex ulmi pumilae solution containing groups showed concentration-dependent increase compared to the control after 24 hours cultivation and the caspase-3 activities were decreased in all Cortex ulmi pumilae solution containing groups compared to the control after 24, 48 and 72 hours cultivation. After 48 and 72 hours cultivation, we could examined the apparent DNA fragmentation in all Cortex ulmi pumilae solution containing groups. In the XTT study, all Cortex ulmi pumilae solution containing groups showed concentration-dependent decrease compared to the control after 24 and 72 hours cultivation but 10% group after 48 hours and 5% and 10% groups after 72hours were presumed statistically significant differences. The expressions of MAP kinase were decreased in all Cortex ulmi pumilae solution containing groups compared to the control after 24, 48 and 72 hours cultivation. Conclusion : From this study we could suggest that Cortex ulmi pumilae be available to the inhibition of apoptosis of human cervical carcinoma cell line in vitro.

• 한방재활의학과학회지
• /
• 제18권4호
• /
• pp.1-11
• /
• 2008

목적 : 뇌허혈은 일시적 혹은, 영구적 뇌동맥 폐색에 의한 뇌혈류의 감소로 유발되며, 허혈 부위에서는 복잡한 병태 생리적 과정을 통하여 신경 세포사가 초래되어 비가역적인 신경학적 손상을 일으킨다. 본 연구에서는 모래쥐를 대상으로 일시적인 전뇌허혈을 유발 시킨 후 해마 치상회에서 허혈로 인한 세포사멸을 관찰하고, 현삼(玄蔘)의 투여가 허혈로 유발된 해마 치상회에서 세포사멸에 미치는 영향과 단기 기억에 미치는 효과를 규명하고자 실험하였다. 연구방법 : 세포사멸은 DNA 분절을 나타내는 terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) 염색법과 단백분해 과정의 마지막 단계에 발현되는 caspase-3에 대한 면역조직화학법을 이용하였고, 단기기억은 step-down avoidance task를 실시하여 평가하였다. 결과 : 본 실험의 결과 일시적인 전뇌허혈은 해마 치상회의 세포사멸을 유의하게 증가시켰으며 단기기억을 감소시켰다. 현삼의 투여는 허혈로 증가된 해마 치상회의 세포사멸을 유의하게 억제하였고, 허혈로 인한 단기 기억의 감소를 유의하게 억제시켰다. 결론 : 본 실험을 통하여 현삼은 뇌허혈로 증가된 세포사멸을 억제하고 단기 기억을 향상시킴을 알 수 있었고, 따라서 현삼은 뇌허혈로 인한 뇌손상을 보호할 수 있는 효과가 있음을 제시하는 바이다.

• 한국식품저장유통학회지
• /
• 제22권5호
• /
• pp.751-757
• /
• 2015

오디 당침출액(MSE)의 산화적 스트레스 개선 효과를 확인하기 위하여 HepG2 세포에 $H_2O_2$로 산화적 스트레스를 유도시킨 다음, MSE의 보호효과를 확인하였다. MSE를 40일간 저장하여 DPPH radical scavensing을 통해 DPPH radical 소거능이 유의적으로 좋았던 저장 40일용 MSE를 선택하여 세포 실험에 적용하였다. HepG2 세포에 $500{\mu}M$ $H_2O_2$를 처리하여 산화적 스트레스를 유발시키고, MSE를 처리하여 세포 생존율을 확인한 결과, MSE 처리로 인한 세포 생존율이 유의적으로 증가하였고, ROS 생성과 과산화물에 대한 지표로 측정된 MDA 농도도 MSE 처리로 인해 효과적으로 억제되었다. 또한, $H_2O_2$ 처리로 감소된 SOD 및 CAT 활성이 MSE 처리로 인해 유의적으로 높아졌으며, $H_2O_2$를 처리로 인한 세포핵의 apoptosis body가 MSE 처리로 인해 감소함을 확인하였으며, 이는 caspase-3 활성 MSE가 억제시킴으로 인해 세포를 보호하고 있음을 확인하였다. 이상의 결과로부터 오디 당침출액은 산화적 스트레스로부터 야기되는 세포독성과 apoptosis로부터 세포 보호 효과를 확인함에 따라 향후 노화와 관련된 다양한 연구소재의 기초 자료 및 질병 예방 소재로의 가능성을 확인하였다.

• 동의생리병리학회지
• /
• 제21권4호
• /
• pp.973-981
• /
• 2007

In the present study, we investigated the antiproliferative activity of the water extract of Samgibopae-tang (SGBPT) in NCI-H460 and A549 non-small-cell lung cancer cell lines. We found that exposure of A549 cells to SGBPT resulted in the growth inhibition in a dose-dependent manner as measured by MTT assay, however SGBPT did not affect the growth of NCI-H460 cells. The antiproliferative effect by SGBPT treatment in A549 cells was associated with morphological changes such as membrane shrinking and cell rounding up. SGBPT treatment did not induce the cell cycle arrest in both cell lines, however the frequency of sub-G1 population was concentration-dependently increased by SGBPT treatment in A549 cells. SGBPT treatment partially induced the expression of tumor suppressor p53 in A549 cells and the expression of cyclin-dependent kinase inhibitor p21(WAF1/CIP1) was markedly increased in both transcriptional and translational levels in A549 cells. The up-regulation of p21 by SGBPT occurred in a similar a concentration dependent manner to that observed with the inhibition of cell viability and induction of sub-G1 population of the cell cycle. However SGBPT treatment did not affect other growth regulation-related genes such as early growth response-1 (Egr-1), nonsteroidal anti-inflammatory drug-activated gene-1 (NAG-1), inducible nitric oxide synthease (iNOS), cyclooxygenases (COXs), telomere-regulatory factors in A549 as well as NCI-H460 cells. Taken together, these findings suggested that SGBPT-induced inhibition of human lung carcinoma A549 cell growth was aoosciated with the induction of p21 and the results provided important new insights into the possible molecular mechanisms of the anti-cancer activity of SGBPT.