• 한국산학기술학회논문지
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• 제18권12호
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• pp.521-528
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• 2017

과도한 음주는 치매 및 알츠하이머 병과 같은 여러 신경계 질환을 일으키는 주요원인 중 하나로 알려져 있으며 이를 해결하기 위한 많은 노력인 진행 중이다. 또한, 홍삼은 신경 세포의 생존, 세포 자멸사의 억제 및 신경 세포의 신경 재생을 향상시키는 것으로 알려져 있다. 본 연구의 목적은 Rg3 풍부 고려홍삼 추출액(KRG)이 알코올 유발성 신경독성으로 인하여 일어나는 PC12 세포의 세포 사멸을 억제 할 수 있는지, 그리고 KRG가 caspase 매개 경로와 관련된 몇 가지 인자들을 어떻게 조절하는지 확인하는 것이다. 그 방법으로, 우리는 PC12 세포에서의 세포 생존율과 세포 사멸율은 EZ-Cytox 세포 생존율 측정 kit와 유세포 분석기로 측정하였고, 세포 자멸 관련 단백질(Bcl-2, Bax, caspase-3)의 발현 정도를 Western blot기법으로 측정하였으며, 측정된 결과의 유의성을 ANOVA 분석법으로 확인하였다. 그 결과, KRG는 Bcl-2의 발현을 증가시키고, Bid와 Bax 및 caspase-3 발현을 저해하였고, 이를 통해 알코올로 유도된 PC12 세포의 세포 사멸을 억제하였다. 이러한 결과를 통해, KRG에 의해 유도된 Bcl-2 발현의 증가와 Bid 및 Bax 발현의 하향 조절이 caspase-3 발현을 하향 조절하고, 결국 미토콘드리아 세포 사멸 경로를 억제한다는 것을 결론내릴 수 있었다. 본 연구는 향 후, KRG가 신경 보호제 후보로서 개발할 가치가 있음을 제시하였다.

• Clinical and Experimental Pediatrics
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• 제62권3호
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• pp.95-101
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• 2019

Purpose: Increased apoptosis was recently found in the hypertrophied left ventricle of spontaneously hypertensive rats (SHRs). Although the available evidence suggests that apoptosis can be induced in cardiac cells by various insults including pressure overload, cardiac apoptosis appears to result from an exaggerated local production of angiotensin in adult SHRs. Altered expressions of Bcl associated X (Bax), Bcl-2, chemokine receptor (CCR)-2, monocyte chemoattractant protein (MCP)-1, transforming growth factor $(TGF)-{\beta}1$, phosphorylated extracellular signal-regulated kinases (PERK), and connexin 43 proteins, and kallikrein mRNA were investigated to explore the effects of losartan on the SHR model. Methods: Twelve-week-old male rats were grouped as follows: control (C), SHR (hypertension: H), and losartan (L; SHRs were treated with losartan [10 mg/kg/day] for 5 weeks). Western blot and reverse transcription polymerase chain reaction assays were performed. Results: Expression of Bax, CCR-2, MCP-1, $TGF-{\beta}1$, PERK, and connexin 43 proteins, and kallikrein mRNA was significantly increased in the H group compared to that in the C group at weeks 3 and 5. Expression of Bax, CCR-2, MCP-1, $TGF-{\beta}1$, and connexin 43 proteins and kallikrein mRNA was significantly decreased after losartan treatment at week 5. PERK protein expression was significantly decreased after losartan treatment at weeks 3 and 5. Bcl-2 protein expression was significantly decreased in the H group compared to that in the C group at weeks 3 and 5. Conclusion: Losartan treatment reduced expression of Bax, CCR-2, MCP-1, $TGF-{\beta}1$, PERK, and connexin 43 proteins, and kallikrein mRNA in SHRs, along with decreased inflammation and apoptosis.

• Archives of Pharmacal Research
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• 제22권5호
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• pp.448-453
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• 1999

In ginsenoside Rh2-treated rat glioma C6Bu-1 cells, apoptotic morphological changes, such as cell shrinkage, chromatin condensation and pyknosis were confirmed by means of electron microscopy. To evaluate whether induction of apoptosis by ginsenoside Rh2 is mediated by the members of Bcl-2 family, we first established C6Bu-1 cells overexpressing Bcl-2. It was demonstrated that the expression of Bcl-2, Bcl-xL, and Bax was not altered in ginsenoside Rh2-treated C6Bu-1 overexpressing C6Bu-1 cells failed to prevent from ginsenoside Rh2-induced cell death. These results suggest the existence of other apoptotic pathway that requires induction of apoptosis by ginsenoside Rh2 rather than the pathway through Bcl-2, $Bcl-x_{L}$ or Bax in C6Bu-1 cells.

• BMB Reports
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• 제36권3호
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• pp.269-274
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• 2003

In the fight against cancer, novel chemotherapeutic agents are constantly being sought to complement existing drugs. Various studies have presented evidence that the apoptosis that is induced by these anticancer agents is implicated in tumor regression, and Bcl-2 family genes play a part in apoptosis following treatment with various stimuli. Here, we present data that a styrylpyrone derivative (SPD) that is extracted from the plant Goniothalamus sp. showed cytotoxic effects on the human breast cancer cell line MCF-7. SPD significantly increased apoptosis in MCF-7 cells, as visualized by phase contrast microscopy and evaluated by the Tdt-mediated dUTP nick end-labeling assay and nuclear morphology. Western blotting and immunostaining revealed up-regulation of the proapoptotic Bax protein expression. SPD, however, did not affect the expression of the anti-apoptotic protein, Bcl-2. These results, therefore, suggest SPD as a potent cytotoxic agent on MCF-7 cells by inducing apoptosis through the modulation of Bax levels.

• Asian Pacific Journal of Cancer Prevention
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• 제13권4호
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• pp.1617-1620
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• 2012

Background: Gold nanoparticles have recently been investigated with respect to biocompatibility according to their interactions with cells. The purpose of this study was to examine cytotoxicity and apoptosis induction by well-characterized gold nanoparticles in human breast epithelial MCF-7 cells. Methods: Apoptosis was assessed by TUNEL, cytotoxicity by MTT assay and caspase 3, 9, p53, Bax and Bcl expression by real-time PCR assays. Results: Gold nanoparticles at up to $200\;{\mu}g/mL$ for 24 hours exerted concentration-dependent cytotoxicity and significant upregulation of mRNA expression of p53, bax, caspase-3 & caspase-9, whereas expression of antiapoptotic bcl-2 was down-regulated. Conclusion: To the best of our knowledge this is the first report showing that gold nanoparticles induce apoptosis in MCF-7cells via p53, bax/bcl-2 and caspase pathways.

• Asian Pacific Journal of Cancer Prevention
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• 제13권5호
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• pp.2095-2100
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• 2012

Sulforaphane (SFN) an isothiocyanate formed by hydrolysis of glucosinolates found in Brassica oleraceae is reported to possess anticancer and antioxidant activities. In this study, we isolated SFN from red cabbage (Brassica oleraceae var rubra) and evaluated the comparative antiproliferative activity of various fractions (standard SFN, extract and purified SFN) by MTT assay in human epithelial carcinoma HEp -2 and and Vero cells. Probable apoptotic mechanisms mediated through p53, bax and bcl-2 were also examined. The SFN fraction was collected by HPLC, enriched for its SFN content and confirmed. Expression of apoptosis-related proteins was detected by western blotting and RT PCR. Results showed that Std SFN and purified SFN concentration found to have closer $IC_{50}$ which is equal to 58.96 microgram/ml (HEp-2 cells), 61.2 microgram/ml (Vero cells) and less than the extract which is found to be 113 microgram/ml (HEp-2 cells) and 125 microgram/ml (Vero cells). Further studies on apoptotic mechanisms showed that purified SFN down-regulated the expression of bcl-2 (antiapoptotic), while up-regulating p53 and Bax (proapoptotic) proteins, as well as caspase-3. This study indicates that purified SFN possesses antiproliferative effects the same as Std SFN and its apoptotic mechanism in HEp-2 cells could be mediated through p53 induction, bax and bcl-2 signaling pathways.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제22권1호
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• pp.15-21
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• 2000

저자는 국소적 재발율에 있어서 큰 차이를 보이는 두 치성종양중 대표적인 함치성 낭종과 법랑아세포종에 있어서 apoptosis 관련 단백(p53, bcl-2, bax)의 발현 양상을 관찰하기 위하여 10례의 함치성 낭종과 16례의 법랑아세포종에 대하여 면역조직화학적 연구를 실시하여 다음과 같은 결론을 얻었다. 1. p53은 함치성 낭종보다 법랑아세포종에서 강하게 발현되었고 법랑아세포종의 조직학적 분류에 따른 p53 발현의 차이를 관찰할 수 없었으며 함치성 낭종의 경우에는 p53이 거의 basal layer에서 발현되었다. 2. Bcl-2는 함치성 낭종의 경우 주로 기저층 혹은 전층에서 발현되었고 법랑아세포종에서는 바깥층 혹은 전층에서 발현되었는데 특히 종양 실질조직중 입방형 또는 주상형 세포층에 집중되어 염색되었다. 3. Bax는 함치성 낭종의 경우 법랑아세포종에 비하여 보다 강하게 발현되었고 함치성 낭종과 법랑아세포종간에 발현양상의 차이는 관찰할 수 없었다. 함치성 낭종 및 법랑아세포종에 있어서 apoptosis 관련단백의 발현양상에 관한 이러한 차이가 법랑아세포종의 독특한 공격적 성장양상과 연관이 있으리라 사료된다.

• Asian-Australasian Journal of Animal Sciences
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• 제18권5호
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• pp.734-740
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• 2005

The blood samples were collected from dairy cows at the same milking stage. The single-strand conformation polymorphism (PCR-SSCP) method was used to analyze for polymorphism at the 5'flanking region of the hsp70 gene. The mRNA expression levels of HSP70, HSF1, Bcl-2 and Bax-$\alpha$ at different daily-mean-temperature were analyzed by relative quantitative RTPCR. The DNA content, cell phase and the ratio of apoptosis of lymphocytes in peripheral blood of dairy cattle at different daily-meantemperature were determined by FCM. The PCR-SSCP products of primer pair 1 showed polymorphisms and could be divided into four genotypes: aa, ab, ac, cc, with the cis-acting element (CCAAT box) included. Mutations in the hsp70 5'flanking region (468-752 bp) had different effects on mRNA expression of HSP70, HSF1, Bcl-2 and Bax-$\alpha$. The ac genotypic cows showed higher expressions of HSP70mRNA, HSF1mRNA and Bcl-2mRNA/Bax-$\alpha$mRNA and lower ratio of apoptosis. These mutation sites can be used as molecular genetic markers to assist selection for anti-heat stress cows.

• 대한두경부종양학회지
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• 제22권1호
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• pp.15-22
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• 2006

Objectives: The salivary gland tumor shows heterogeneity in histologic patterns and biological behavior. The aim of this study is to elucidate the relationships between apoptosis and expressions of apoptosis-related factors(bcl-2, bax, M30), p53 and MIB-1 in the salivary gland tumors. Methods: Immunohistochemical stains for apoptosis-related factors, p53 and MIB-1 and TUNEL study for apoptosis were performed in 46 cases of salivary gland tumors 02 benign and 34 malignant). Results: Twenty(43.5%) of 46 cases showed positive reaction for apoptosis, and the expression rates of bcl-2, bax, M30, p53 and MIB-1 were 85.3%, 68.8%, 65.9%, 39.1% and 26.1%, respectively. A significant difference between benign and malignant tumors was only noted in MIB-1 expression(p=0.0167). In malignant tumors, apoptosis showed no significant relationships to expressions of apoptosis-related factors. There were inverse relationships between p53 and bcl-2 expression(p=0.0375), and between M30 and MIB-1 expressions(p=0.0379). No significant differences of apoptosis, bcl-2, bax, M30, p53 and MIB-1 expression rates according to the tumor size, lymph node status, recurrence and survival were found. Conclusion: In the development of benign and malignant salivary gland tumors, apoptosis might be associated, however, apoptosis and expressions of apoptosis-related factors seemed to be not reliable prognostic factors in malignant salivary gland tumors.

• Clinical and Experimental Reproductive Medicine
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• 제30권1호
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• pp.77-84
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• 2003

Objective : The aim of this study was to evaluate the influence of three different media on preimplatation embryo development and the expression of Bcl-2, Mcl-1, Bax, and Bok in mouse. Materials and Methods: Two-cell embryos were retrieved from ICR female mice (4 weeks old) at 48 hr after hCG injection and cultured in Ham's F-10, HTF, and G1.2 media. The developmental rate of 2-cell embryos was evaluated from 24 hr to 72 hr after culture. RT-PCR was performed for the detection of Bcl-2, Mcl-1, Bax, and Bok gene expression. Results: The rates of morula and blastocyst in HTF and G1.2 media (88%, 98.1%) were significantly higher than those in Ham's F-10 media (39.6%) at 48 hr. Likewise, the rates of hatching and hatched blastocyst in HTF and G1.2 media (21.9%, 52.9%) were higher than those in Ham's F-10 media (3.5%) at 72 hr. Bcl-2 and Bax mRNAs were highly detected in embryos cultured in Ham's F-10 when compared in embryos cultured in HTF and G1.2. In contrast, the expression of Mcl-1 and Bok was not significantly different. Conclusion: These results show that HTF and G1.2 culture media increase the rate of blastocyst formation and stimulate Bcl-2 and Bax gene expression in mouse preimplantation embryos.