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

• 대한한의학회지
• /
• 제19권1호
• /
• pp.145-164
• /
• 1998

The purpose of this study is to evaluate the effect of Injinchunggantang-derivative on proliferation of hepatocyte in rats. Cell viability is studied by MTI assay. The gene related to cell replication such as p53, waf1, bcl-2 and $bcl-_{X_L}$ is quantitized by quantitative RT-PCR and the proteins coded by these genes are studied by Western blotting. The results are as follows. 1. The hepatocytes cultured in medium with lnjinchunggantang-derivative showed better viability compared with control grroup in MTI assay, and the hepatocytes cultured in medium with the Injinchunggantang-derivative-and-ethanol-mixed group showed better viability than the hepatocytes cultrued in 10% ethanol culture medium(control group), noting that Injinchunggantang-derivative has protective effect on hepatocyte injury. There was no dose- and time-dependence. 2. In quantitative RT-PCR, i) Bel-2 gene increased significantly both in Injinchunggantang-derivative group and in Injinchunggantang-derivative-and-ethanol-mixed group, while it showed no significant increase or decrease in other group. ii) $Bcl-_{X_L}$ gene increased significantly in Injinchunggantang-derivative group as well as in Injinchunggantang-deri vative-and-ethanol -mixed group. iii) P53 gene showed no significant increase or decrease in hepatocytes cultured in medium with 10% ethanol and in hepatocytes cultured in medium with Injinchunggantang-derivative-and-ethanol-mixed group, suggesting that 10% ethanol induced cell toxicity, thus increased p53 gene expression. iv) Wafl gene showed no significant increase or decrease in hepatocytes cutured in medium with Injinchtrnggantang-derivative, while increased in hepatocytes cultured in medium with 10% ethanol and in hepatocytes cultured in medium with Injinchtrnggantang-derivative-andethanol-mixed group, suggesting that 10% ethanol induced cell toxicity increased wafl gene expression. 3. In the study on protein by western blotting, the band of bcl-2 and $bcl-_{X_L}$ were widened in Injinchtrnggantang-derivative group. Especially the amount of $bcl-_{X_L}$ increased significantly compared with other groups. But in the study on p53 and wafl, there was no significant difference among those groups. Above study shows that Injinchunggantang-derivative has good effect on cell viability and that the genes resistant to cell death such as bcl-2 and $bcl-_{X_L}$ are induced by Injinchunggantang-derivative to resist to cell death by toxic agent And this is reconfirmed in protein study using' western blotting: These results suggest that Injinchunggantang-derivative has inhibitory effect on cell death as well as protective effect on hepatocyte. Therefore this prescription is recommended in various liver diseases such as chronic liver disease and-induced hepatic injury.

• 대한의생명과학회지
• /
• 제14권3호
• /
• pp.157-165
• /
• 2008

Recently, the number of patients with prostate cancer has been increased gradually by both the change of living environment and the increase of aged population. In this study the serum prostate specific antigen (PSA) level was compared to the Gleason score known as a prognostic factor for the prostate cancer. In the Gleason score 6 and $9{\sim}10$, the average age was 69.68 years old and 69.52 years old, respectively and there was no statistically difference in both of age and Gleason score. the PSA serum consistency appeared <4 ng/mL as example 1, $4{\sim}20ng/mL$ as example 17 and ${\geq}20ng/mL$ as example 4 in the Gleason score 6, and In the Gleason score $9{\sim}10$, it appeared <4 ng/mL as example 1, $4{\sim}20ng/mL$ as example 6 and ${\geq}20ng/mL$ as example 15. PSA serum consistency in the Gleason score $9{\sim}10$ showed higher value than those of Gleason score 6 (P<0.05). Next, expression ratios of bcl-2, Ki-67 and p53 were examined in the Gleason score 6 and $9{\sim}10$. the p53 expression ratio, a tumor suppression gene, appeared the significance statistically by the classification of the Gleason score as example 7 (28%) in the Gleason score 6 and as example 16 (64%) in the Gleason score $9{\sim}10$ (P<0.05). but not different in the expression ratios of the Ki-67 and bcl-2. The expression ratio of p53 by the expression ratio of bcl-2 and the expression ratio of Ki-67 by the expression ratio of p53 had a positive relationship in all of the Gleason score 6 and the Gleason score $9{\sim}10$ (P<0.05). However, the expression ratio of Ki-67 by the expression ratio of bcl-2 did not show any significance in the Gleason score $9{\sim}10$ (P<0.05). Therefore, the results suggested that p53 expression could be used as an independent prognostic factor.

• 대한한방내과학회지
• /
• 제35권1호
• /
• pp.79-91
• /
• 2014

Objectives : To investigate the anti-cancer effect of Palbohoichoon-tang (PBHCT) extracts. Methods : The cell viability was assessed by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MMT) assay and cell morphological changes were microscopically analyzed after staining with $10{\mu}M$ 2-[4-amidinophenyl]-6-indolecarbamidine dihydrochloride (DAPI) and TUNEL. We also analyzed expression of Bcl2, $Bcl_{xL}$, Bax, procaspase-3, procaspase-9, and procyclic acidic repetitive protein (PARP) by western blot method. Results : Observations showed that PBHCT induced the apoptotic cell death proved by increased sub-G1 phase cell population, apoptotic body formation and chromatin condensation. Western blot analysis of total cell lysates revealed that the PBHCT induced cleavage of caspase-9, caspase-3 and poly (ADP-ribose) polymerase (PARP). In addition, PBHCT dose-dependently increased the activity of caspase-9, caspase-3 and PARP-1. Furthermore, PBHCT reduced anti-apoptotic Bcl2, $Bcl_{xL}$ expression which contributed to the loss of mitochondrial membrane potential and the activations of caspase-9 and caspase-3. Conclusions : These findings suggest that PBHCT exerts anti-cancer effects on human neuroblastoma SH-SY5Y cells by inducing apoptotic death via down-regulation of anti-apoptotic proteins such as Bcl2 and $Bcl_{xL}$, up-regulation of pro-apoptotic proteins such as Bax, and activation of caspase cascades and PARP-1.

• Molecules and Cells
• /
• 제41권7호
• /
• pp.684-694
• /
• 2018

Upregulation of human telomerase reverse transcriptase (hTERT) expression is an important factor in the cellular survival and cancer. Although growing evidence suggests that hTERT inhibits cellular apoptosis by telomere-independent functions, the mechanisms involved are not fully understood. Here, we show that hTERT contains a BH3-like motif, a short peptide sequence found in BCL-2 family proteins, and interacts with anti-apoptotic BCL-2 family proteins MCL-1 and BCL-xL, suggesting a functional link between hTERT and the mitochondrial pathway of apoptosis. Additionally, we propose that hTERT can be categorized into the atypical BH3-only proteins that promote cellular survival, possibly due to the non-canonical interaction between hTERT and antiapoptotic proteins. Although the detailed mechanisms underlying the hTERT BH3-like motif functions and interactions between hTERT and BCL-2 family proteins have not been elucidated, this work proposes a possible connection between hTERT and BCL-2 family members and reconsiders the role of the BH3-like motif as an interaction motif.

• 생명과학회지
• /
• 제17권4호
• /
• pp.552-560
• /
• 2007

본 연구에서는 인체백혈병세포 U937에서 HDAC 저해제에 의한 증식억제, 세포주기 교란 및 apoptosis 유도에 미치는 Bcl-2 유전자의 영향에 관하여 조사하였다. 이를 위하여 U937/vector 및 U937/Bcl-2 세포주를 대상으로 대표적인 HDAC 저해제인 TSA 및 Na-B 처리에 의한 세포 증식 및 생존율에 미치는 영향을 조사한 결과, TSA에 의한 U937 세포의 증식억제 및 생존율의 감소는 Bcl-2의 과발현에 의하여 차단되는 효과를 보였으나, Na-B는 U937/vector 및 U937/Bcl-2세포사이에 큰 변화를 보이지는 않았다. 세포주기 교란효과에서 Na-B는 TSA에 비하여 유의적인 차이를 보이지 못하였으며, 이는 TSA에 의한 apoptosis가 U937/Bcl-2 세포에서는 억제되었으나, Na-B에 의한 apoptosis는 Bcl-2의 과발현에 의하여 차단되지 못한 것과 연관성이 있는 결과였다. 또한 TSA에 의한 apoptosis 유발의 Bcl-2에 의한 차단 효과는 TSA에 의하여 활성화된 caspase의 활성 억제, Bcl-2 발현 자체의 완화 등 apoptosis 조절 인자들의 발현 및 활성 변화에 기인 된 것임을 알 수 있었다.

• Molecules and Cells
• /
• 제46권7호
• /
• pp.420-429
• /
• 2023

Age-related macular degeneration (AMD) is one of the leading causes of blindness in elderly individuals. However, the currently used intravitreal injections of anti-vascular endothelial growth factor are invasive, and repetitive injections are also accompanied by a risk of intraocular infection. The pathogenic mechanism of AMD is still not completely understood, but a multifactorial mechanism that combines genetic predisposition and environmental factors, including cellular senescence, has been suggested. Cellular senescence refers to the accumulation of cells that stop dividing due to the presence of free radicals and DNA damage. Characteristics of senescent cells include nuclear hypertrophy, increased levels of cell cycle inhibitors such as p16 and p21, and resistance to apoptosis. Senolytic drugs remove senescent cells by targeting the main characteristics of these cells. One of the senolytic drugs, ABT-263, which inhibits the antiapoptotic functions of Bcl-2 and Bcl-xL, may be a new treatment for AMD patients because it targets senescent retinal pigment epithelium (RPE) cells. We proved that it selectively kills doxorubicin (Dox)-induced senescent ARPE-19 cells by activating apoptosis. By removing senescent cells, the expression of inflammatory cytokines was reduced, and the proliferation of the remaining cells was increased. When ABT-263 was orally administered to the mouse model of senescent RPE cells induced by Dox, we confirmed that senescent RPE cells were selectively removed and retinal degeneration was alleviated. Therefore, we suggest that ABT-263, which removes senescent RPE cells through its senolytic effect, has the potential to be the first orally administered senolytic drug for the treatment of AMD.

• 대한한의학방제학회지
• /
• 제31권4호
• /
• pp.253-264
• /
• 2023

Objectives : Bambusae Caulis in Liquamen (BCL), a traditional herbal medicine, is a distilled product of condensation from the burning of fresh bamboo stems. We previously identified the anti-oxidant capacity of BCL in hepatocytes and suggested that BCL is a promising therapeutic candidate for treating oxidative stress-induced hepatocellular damage. Despite the importance of the role played by Kupffer cells in liver disease, the efficacy of BCL on Kupffer cells is unclear. Therefore, this study aimed to determine whether BCL could suppress LPS-induced inflammation and LPS+ATP-induced inflammasomes in Kupffer cells. Methods : We used ImKCs, a murine immortalized Kupffer cell line to examined whether BCL inhibited LPS-induced inflammation response and oxidave stress. And, we prepared a total of 18 L of BCL, purchased from Bamboo Forest Foods Co., Ltd. (648 Samdari, Damyang-eup, Damyang-gun, Jeollanam-do, Republic of Korea), was concentrated using a decompression concentrator. Result : The LPS-induced release of inflammatory cytokines was abolished by BCL treatment. Also, BCL treatment suppressed the LPS+ATP-induced expression of inflammasome proteins (NLRP3, IL-1, and IL-18), and inhib β ited the release of IL-1 . BCL decreased LPS-or LPS+ATP-induc β ed reactive oxygen species production. In addition, BCL increased nuclear translocation of Nrf2 and the expression of HO-1 in a time-dependent manner. Conclusion : These results suggest the efficacy of BCL with respect to its anti-inflammatory and anti-inflammasome effects mediated by Nrf2 in Kupffer cells.

• 대한한방부인과학회지
• /
• 제24권2호
• /
• pp.33-51
• /
• 2011

Objectives: This study was performed to assess the protective effect of Saengmaek-san (SM) on UVB-induced HaCaT cell damage. Methods: The protective effects of Saengmaek-san(SM) were determined by UVB-induced HaCaT assay. We assessed protective effects of Saengmaek-san (SM) on LDH release and nitrite release from HaCaT. And COX-2, Bcl-2, Bax, $TNF{\alpha}$, c-jun, c-fos, NF-kB, iNOS, Bcl-xL gene expression were determined in HaCaT using real-time PCR method. Results: 1. SM inhibited LDH-release, nitrite production in UVB-exposed HaCaT. 2. SM suppressed the gene expression of COX-2, $TNF{\alpha}$ in UVB-exposed HaCaT. 3. SM increased the gene expression of Bcl-2, Bax, Bcl-xL family protein in UVB-exposed HaCaT. 4. SM suppressed the gene expression of c-jun, c-fos, NF-kB in UVB-exposed HaCaT. Conclusions: The study showed SM inhibited the cell damage in UVB-exposed HaCaT.

• 한국식품저장유통학회지
• /
• 제24권7호
• /
• pp.992-999
• /
• 2017

대나무 죽력의 ACE 저해효과와 마우스모델에서의 혈행개선 효과를 보았다. 혈전용해능에서 청국장추출물은 2.56 unit, 미정제죽력에서 4.71 unit, 정제죽력에서 4.32 unit를 나타내어 모두가 plasmin 보다 유의하게 높은 활성이 높고, ACE 활성 저해 효과에서는 죽력 원액, 미정제 죽력과 정제 죽력에서 각각 70.25%, 69.01%, 68.23%로 대조군으로 사용된 청국장이 26.56%으로 나타나 높은 억제효과를 보였다. 마우스모델시험에서 고지방식이군과 함께 고지방식이군+정제죽력 0.01%와 0.05%를 투여, 혈청 중 지질변화에서 triglyceride의 함량은 고지방식이군이 일반식이군(271.1 mg/dL)에 비해 314.4 mg/dL로 증가하였고, 고지방식이+죽력 0.01%군은 301.5 mg/dL로 나타났으며, 고지방식이+죽력 0.05%군은 289.2 mg/dL으로 유의하게 중성지질이 감소를 나타내었고, total-cholesterol 함량은 327.2 mg/dL로 증가하였으며, 고지방식이+죽력 0.01%군은 311.9 mg/dL로 나타났으며, 고지방식이+죽력 0.05%군은 293.7 mg/dL로 고지방식이군에 비교, 유의하게 total-cholesterol이 감소세를 나타내었다. 한편 HDL-cholesterol은 일반식이군이 206.0 mg/dL로 가장 높은 값을 나타내었으며, 고지방식이+죽력 0.01%군 196.6 mg/dL, 고지방식이+죽력 0.05%군 189.2 mg/dL, 고지방식이군 162.2 mg/dL순으로 나타났고, 죽력첨가군 0.01%군과 0.05%군간의 유의성은 없었다. 혈류개선효과에서는 고지방식이에 비하여 고지방식이+죽력 0.05% 군이 고지방식이군, 고지방식이+죽력 0.01%군보다 모두 유의하게 빠르게 모세관을 통과하여 빠른 혈액 유동성을 나타내었다. 혈소판응집 억제능에서는 고지방식이군의 경우 대조군인 일반식이군에 비해 혈소판 응집이 증가한 반면 고지방식이+죽력 0.01%군과 고지방식이+죽력 0.05%군은 고지방식이군에 비해 혈소판응집이 억제되는 것으로 나타났다.