• 한방재활의학과학회지
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• 제20권1호
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• pp.1-12
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• 2010

Objectives : Cool Pharmacopuncture for static blood is a famous pharmacopuncture treatment that treated disease caused by static blood. Hyolhae(Sp10) is also a famous point of acupuncture that treated that. This study was designed to evaluate the effects of Cool Pharmacopuncture for static blood into Hyolhae(Sp10) on BAX and BCL-2 expression in the experimental traumatic brain injury(TBI) rats. Methods : Male rats were divided into 3 groups. I was no treatment after TBI. II was treatment with needle-prick acupuncture after TBI. III was treatment with Cool Pharmacopuncture for static blood into Hyolhae(Sp10) after TBI. The author carried out neurological motor behavioral test, histological assessment test. Neurological motor behavior tests consist of rotarod test, beam-walking test and postural reflex test. In the histological assessment test, BAX and BCL-2 expression, hematoxylin & eosin staining were experimented. Results : In neurological motor behavior tests, motor and cognitive function recovery was significantly increased in the II, III as compared with I (p<0.05). Especially III was significantly increased as compared with II (p<0.05). BAX expression was significantly decresed in order of the III, II, I after 7 and 14 days later. BCL-2 expression was investigated in the III, II as compared with I. Especially Most incresed expression was experimented in the III. Conclusions : According to the above results, Cool Pharmacopuncture for static blood can inhibit apoptosis of cells after TBI in rats by contol of BAX and BCL expression.

• Asian Pacific Journal of Cancer Prevention
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• 제15권17호
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• pp.7065-7068
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• 2014

To investigate the anti-proliferative mechanism of mangiferin in a human nasopharyngeal carcinoma cell line, CNE2 cells were incubated with different concentrations of mangiferin (12.5, 25, 50, 100, 150 and $200{\mu}M$) or with PBS as a control for 72 hours. Analyses were made of the cell cycle and apoptosis with measurement of mRNA and protein levels of two apoptosis-related genes, Bcl-2 and Bax. Flow cytometry assays showed mangiferin could inhibit CNE2 cell proliferation via G2/M arrest and induction of early apoptosis. Real time PCR and Western blotting showed the mRNA and protein level of Bcl-2 to be down-regulated, while those of Bax were upregulated, when CNE2 cells were treated with mangiferin. This investigation indicated anti-proliferation effects of mangiferin through induction of cell apoptosis regulated by Bcl-2 and Bax expression.

• 생명과학회지
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• 제20권9호
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• pp.1409-1414
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• 2010

이 연구는 실험용 쥐를 대상으로 운동에 따라 골격근에서 일어나는 세포고사 경로 중 내인성 경로와 외인성경로에 의해 이루어지는 세포고사의 신호기전을 확인하여 운동이 세포고사에 어떠한 영향을 미치는지에 대한 구명하는데 있다. 이를 위해 ICR계 수컷 흰쥐 20마리를 일주일의 적응기간을 거쳐 통제집단(CON: n=10)과 운동집단(EX: n=10)으로 배정하였다. 운동은 8주간 주 5회 실시하였고, 트레드밀 속도 16.4 m/min와 경사도 4%로 설정하여 40분간 지속적인 운동을 실시하였다. 세포고사의 신호 경로 중 내인성 경로에 대한 검증 결과 Bcl-2, Bax 단백질, 그리고 Bcl-2/ Bax ratio는 그룹간 통계적 유의성은 나타나지 않았다. 반면, 세포고사의 경로 중 외인성 경로에 대한 검증 결과 caspase-8 단백질의 발현은 운동집단이 통제집단보다 유의하게 낮은 것으로 나타났으며(p<0.05), 세포고사 억제 단백질인 HSP70 단백질 발현은 운동집단이 통제집단보다 높게 나타났다. 더욱이, 세포고사 최종인자인 caspase-3의 활성화는 이루어 지지 않은 것으로 관찰되었다. 따라서 세포사멸의 신호경로 중 내인성 경로에 작용하는 Bcl-2와 Bax보다는 외인성 경로인 caspase-8과 HSP70의 영향으로 caspase-3가 분할되지 못하여 세포고사가 일어나지 않은 것으로 사료된다.

• Asian Pacific Journal of Cancer Prevention
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• 제14권8호
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• pp.4599-4602
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• 2013

We intended to study the mechanism of the inhibitory action of curcumin on human non-small cell lung cancer A549 cell. The cell growth was determined by CCK-8 assay, and the results indicated that curcumin inhibited the cell proliferation in a concentration dependent manner. And to further confirm the relative anti-cancer mechanism of curcumin, RT-PCR was carried out to analysis the expression of relative apoptotic proteins Bax, Bcl-2. We found that curcumin could up-regulate the expression of Bax but down-regulate the expression of Bcl-2 in A549 cells. In addition, curcumin affect the mitochondrial apoptosis pathway. These results suggested that curcumin inhibited cancer cell growth through the regulation of Bcl-2/Bax and affect the mitochondrial apoptosis pathway.

• 약학회지
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• 제43권3호
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• pp.385-390
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• 1999

Camptothecin (CPT) has been known to induce apoptosis in various cancer cell lines. To examine the intracellular apoptotic death signal initiated by CPT, we investigated the possible connection between caspase-3 activation and GSH depletion during CPT-induced apoptosis in HL-60 cells. Treatment of cells with $1{\;}{\mu}M$ CPT induced PARP cleavage accompanied by DNA fragmentation. z-VAD-fmk, a caspase-3 inhibitor, blocked the CPT-induced DNA fragmentation. Pretreatment of cells with N-acetylcysteine, a precursor of GSH biosynthesis, failed to inhibit CPT-induced PARP celavage and DNA gragmenatation. No significant changes in GSH depletion is not essential for caspase activation during CPT-induced apoptosis. We also investigated whether CPT-induced apoptosis is associated with changes of the levels of Bax and Bcl-2, two proteins involved in the control of apoptosis. Bcl-2 levels exhibited a late decrease compared with the kinetics of DNA fragmentation, whereas Bax levels increased more rapidly after CPT treatment. These results suggest that Bax plays more important role than Bcl-2 in inducing DNA fragmentation and may function upsteam of proteolytic activation of caspase-3 pathway in CPT-induced apoptosis.

• 동아시아식생활학회지
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• 제24권6호
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• pp.739-747
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• 2014

본 연구는 인체유방암세포인 MDA-MB-231세포를 이용하여 IDE를 처리하였을 때 세포사멸에 미치는 영향을 세포화학적 방법으로 확인하였다. IDE를 각각 0, 20, 30 및 $40{\mu}g/mL$ 첨가하여 24시간 배양한 후, 세포증식 억제, 막투과성, 세포내 ROS 분석 및 세포사멸 단계의 특성을 FACS 분석하고, RT-PCR에 의한 사멸관련 유전자 중 Bax/Bcl-2 ratio 분석을 통하여 IDE의 항암작용의 조절작용을 밝히고자 하였다. MTT의 세포 증식 억제는 첨가된 IDE의 농도에 따라 유의적으로 감소하였다(p<0.05). 이와 동시에, trypan blue에 대한 염색성과 DCF-DA 형광 분석의 결과는 각각, 막투과성과 세포내 ROS 농도가 모두 농도 의존적으로 증가됨을 보였다(p<0.05). 이와 같은 IDE 농도에 따른 세포화학적 변화 중에서 세포의 초기사멸에서 후기사멸 과정으로 급격한 사멸단계의 변화가 특히, IDE 농도 30과 $40{\mu}g/mL$에서 일어났다. RT-PCR 분석에 의한 Bax/Bcl-2 ratio도 IDE 농도 30과 $40{\mu}g/mL$에서 급격히 증가하였다(p<0.05). 이와 같은 세포화학적 결과와 RT-PCR 결과를 종합해 볼 때, IDE의 유방암세포(MDA-MB-231)에 대한 세포사멸작용은 막 투과성의 증가와 ROS 증가를 통하여 세포에 점진적인 손상을 일으키며, 이는 세포의 생화학적 변화도 초래하여 세포 증식 억제를 감소시켜, 결국 세포내의 사멸관련 유전자 지표인 Bax/Bcl-2 ratio를 크게 변화시키는 일련의 세포사멸을 점진적으로 유도시켜 항유방암의 효과의 가능성을 제시하였다.

• 한방재활의학과학회지
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• 제19권3호
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• pp.1-13
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• 2009

Objectives : This study evaluates neuroprotective effect of Gastrodiae Rhizoma on apoptosis in the cerebral infarct. Methods : Cerebral infarct was induced by the middle cerebral artery occlusion (MCAO) for 2 hours with intraluminal thread method in Sprague-Dawley rats. Then ethanol extract of Gastrodiae Rhizoma was administered orally for 3 days. Infarct area and volume were evaluated with TTC staining. Neuronal apoptosis was identified with TUNEL labeling. Apoptosis modulatory effect was observed with immunohistochemical Bax, Bcl-2, iNOS, and MMP-9 expressions in penumbra. Results : 1. Ethanol extract of Gastrodiae Rhizoma reduced infarct size partly and volume significantly of in the MCAO rat brain. 2. Ethanol extract of Gastrodiae Rhizoma reduced TUNEL positive cell ratio in the penumbra of MCAO rat brain significantly. 3. Ethanol extract of Gastrodiae Rhizoma suppressed Bax, iNOS and MMP-9 expression in the penumbra of MCAO rat brain significantly. 4. Ethanol extract of Gastrodiae Rhizoma did not change Bcl-2 expression in the penumbra of MCAO rat brain. But expression ratio of Bcl-2 against Bax was increased in the Gastrodiae Rhizoma group. Conclusions : These results suggest that Gastrodiae Rhizoma plays an anti-apoptotic neuroprotective effect through suppression of Bax, iNOS, and MMP-9 expressions and relative up-regulation of Bcl-2 in the ischemic brain tissue.

• Clinical and Experimental Reproductive Medicine
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• 제27권3호
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• pp.245-251
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• 2000

Objective: To investigate the distribution of BCL-2, BAX proteins and DNA fragmented cells in the normal human endometrium during at each menstrual cycle in order to find out whether apoptosis regulates cyclic endometrial change. Methods: Normal endometrial tissues were obtained from 40 patients, $32{\sim}45$ year of age, all with regular menstrual cycle, who were undergoing abdominal hysterectomy for myoma of uterus or cervical intraepithelial neoplasia for the period from 1992 through 1997. Immunohistochemical staining was used to determine the expression of BCL-2 and BAX protein with paraffin-embedded tissues. Results: BCL-2 was expressed on the glandular epithelial cells and stromal cells during the proliferative phase. The intensity of BCL-2 was increased predominantly on the basal layer than the functional layer in late proliferative phase. However, BCL-2 immunoreactivity was decreased in the secretory phase. BAX was expressed predominantly during the secretory phase. The intesity was increased in late secretory phase rather than early secretory phase. DNA fragmented cells were detected in a few cells at each phase. However, it was increased during the late secretory phase. Conclusion: Apoptosis-related genes, BCL-2 and BAX, may play a role in the regulation of cyclic endometrial change.

• 대한한방부인과학회지
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• 제23권3호
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• pp.38-55
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• 2010

Purpose: This study was designed to find out the anti-cancer effects of Samultang-Gami which was composed of Rehmanniae Radix(RR), Angelicae Gigantis Radix(AGR), Cnidii Rhizoma(CR), Paeoniae Radix(PR), Cortex Moutan Radicis(CMR), Hedyotis Diffusa(HD) and Caesalpinia Sappan on HeLa, HepG2 and AGS cells. Methods: Various cancer cell lines including HeLa, HepG2 and AGS cells, were used. In vitro anti-cancer effects were measured by MTT assay using cancer cell lines treated with various concentrations of 70% ethanol extract of Samultang-Gami. Expression of cell cycle arrest mediators including Bax, Bcl-2, p53 and DARP-1 proteins were measured by Western blot analysis. Results: 1. Samultang-Gami decreased the viability of HeLa and HepG cells in a dosedependent manner. 2. AGR, CMR, PR and HD decreased the viability of HeLa, HepG2 and AGS cells. 3. We could observe that the decreased Bax and Bcl-2 expression level and the increased PARP-1 expression level by Samultang-Gami extracts treated in HeLa cells. 4. We could observe that the decreased Bcl-2 expression level and the increased Bax, p53 and PARP-1 expression level by RR extracts treated in HeLa cells. and also could observe that the reduction of the protein level of Bcl-2, p53 and PARP-1 and the increase of the protein level of Bax by PR in HeLa cells. 5. We could observe that the increased p53 expression level, the decreased PARP-1's that and the unchanged Bax and Bcl-2's that by Samultang-Gami extracts treated in HepG2 cells. 6. We could observe that the reduced Bcl-2 expression level by each of RR extracts and PR extracts in HepG2 cells. 7. The treatment of Samultang-Gami in AGS cells didn't have any effect on the expression level of Bax, Bcl-2, p53 and PARP-1. 8. We could observe that the increased p53 and PARP-1 expression level by each of CR, RR and PR extracts in AGS cells. Conclusion: Taken together, we suggest that Samultang-Gami exhibits cytotoxic effects on HeLa, HepG2 and AGS cells, causing apoptosis. The results showed that Samultang-Gami may do so by regulating the expression of specific target molecules that promote efficient apoptotic cell death in a dose-dependent manner.

• Asian Pacific Journal of Cancer Prevention
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• 제17권11호
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• pp.5031-5035
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• 2016

Docetaxel, recognized as a stabilizing microtubule agent, is frequently administrated as a first line treatment for prostate cancers. Due to high side effects of monotherapy, however, combinations with novel adjuvants have emerged as an alternative strategy in cancer therapy protocols. Here, we investigated the combined effects of stattic and docetaxel on the DU145 prostate cancer cell line. Cytotoxicity was evaluated by MTT assay. To understand molecular mechanisms of stattic action, apoptotic related genes including Bcl-2, Mcl-1, Survivin and Bax were evaluated by real-time RT-PCR. Alteration in the expression of pro-apoptotic Bax and anti-apoptotic Bcl-2 genes and Bax/Bcl-2 ratio were investigated via the $2^{{\Delta}{\Delta}CT}$ method. The $IC_{50}$ values for docetaxel and stattic were $3.7{\pm}0.9nM$ and $4.6{\pm}0.8{\mu}M$, respectively. Evaluation of key gene expression levels revealed a noticeable decrease in antiapoptotic Bcl-2 and Mcl-1 along with an increase in pro-apoptotic Bax mRNA levels (p<0.05). Our results suggest that combination of a STAT3 inhibitor with doctaxel can be considered as a potent strategy for induction of apoptosis via increasing Bax mRNA expression.