• The Korean Journal of Physiology and Pharmacology
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• 제7권4호
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• pp.199-205
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• 2003

Osteoblasts are affected by TNF-${\alpha}$ overproduction by immune cells during inflammation. It has been suggested that functional $NF-{\kappa}B$ sites are involved in TNF-${\alpha}$-induced bone resorption. Thus, we explored the effect of pyrrolidine dithiocarbamate (PDTC), which potently blocks the activation of nuclear factor $(NF-{\kappa}B)$, on the induction of TNF-${\alpha}$-induced activation of JNK/SAPK, AP-1, cytochrome c, caspase and apoptosis in MC3T3E1 osteoblasts. Pretreatment of the cells with PDTC blocked TNF-${\alpha}$-induced $NF-{\kappa}B$ activation. TNF-${\alpha}$-induced activation of AP-1, another nuclear transcription factor, was suppressed by PDTC. The activation of c-Jun N-terminal kinase, implicated in the regulation of AP-1, was also down regulated by PDTC. TNF-${\alpha}$-induced apoptosis, release of cytochrome c and subsequent activation of caspase-3 were abolished by PDTC. TNF-${\alpha}$-induced apoptosis was partially blocked by Ac-DEVD-CHO, a caspase-3 inhibitor, suggesting that caspase-3 is involved in TNF-${\alpha}$-mediated signaling through $NF-{\kappa}B$ in MC3T3E1 osteoblasts. Thus, these results demonstrate that PDTC, has an inhibitory effect on TNF-${\alpha}$-mediated activation of JNK/SAPK, AP-1, cytochrome c release and subsequent caspase-3, leading to the inhibition of apoptosis. Our study may contribute to the treatment of TNF-${\alpha}$-associated immune and inflammatory diseases such as rheumatoid arthritis and periodontal diseases.

• Imaging Science in Dentistry
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• 제35권3호
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• pp.147-156
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• 2005

Purpose : To observe the histopathological changes and caspase-3 expression in the submandibular gland in streptozotocin-induced diabetic rats after irradiation. Materials and Methods : The male Sprague-Dawley rats weighing approximately 250 gm were divided into four groups: control, diabetes, irradiation, and diabetes-irradiation groups. Diabetes mellitus was induced in the rats by injecting streptozotocin. Rats in the control and irradiation groups were injected with citrate buffer only. After 5days, rats in irradiation and diabetes-irradiation groups were irradiated with a single absorbed dose of 10 Gy to the head and neck region. All the rats were sacrificed at 3, 7, 14, 21, and 28 days after irradiation. The specimen including the submandibular gland were sectioned and observed using histopathological and immunohistochemical methods. Results : In the irradiation group, the condensed nucleus, karyolysis, and degeneration of the acinar cells and atrophy of the duct cells were observed in the early experimental phase. However, the acinar cells were found to be normal at 28 days after irradiation. In the diabetes group, the condensed nucleus, karyolysis, atrophy, and degeneration of the acinar cells were observed in the early experimental phase. However, the acinar cells were found to be normal at 21 days after diabetic state induction. In the diabetes-irradiation group, the ductal epithelial cells were predominant in their glandular tissues at 28 days after irradiation. In all of the experimental groups, the most prominent change of the acinar cells and ductal cells were observed at 14 days after diabetic state induction and irradiation. Conclusion The expression of caspase-3 in the acinar cells and ductal cells of the submandibular gland was weak after irradiation, but that in the acinar cells, ductal cells, and fibrous cells of the submandibular gland was prominent after diabetic state induction.

• 대한두경부종양학회지
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• 제28권2호
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• pp.107-113
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• 2012

배 경 편평상피암은 두경부의 악성종양 중 가장 흔하며, 임상적인 경과가 불량하다. 따라서 나쁜 예후를 가지는 환자군을 조기에 선별하여 더 적극적인 치료의 시행을 결정짓는 표지자의 필요성이 대두된다. 우리는 일련의 두경부 편평세포암 검체에서 몇몇 분자 표지자의 예후적 유용성을 평가하고자 하였다. 방 법 23예의 두경부 편평세포암 검체를 대상으로 VEGF, p53, Apaf-1, caspase-9의 발현과 몇몇 임상병리학적 지표들간의 연관성을 면역조직화학염색을 통해 조사하였다. 결 과 환자군은 남성이 더 많았으며 평균연령은 63.7세였다. 1기가 5예, 2기가 2예, 3기가 8예, 4기가 8예였다. 평균생존기간은 37.3개월이었다. VEGF단백 발현은 종양의 크기와 통계적으로 유의한 연관성을 보였다. 이와 더불어 VEGF단백 발현은 병기, 그리고 림프관 침습과 연관적인 경향성을 보였다. 그러나 VEGF단백 발현과 생존기간과는 관련성이 없었다. 또한, Apaf-1과 caspase-9의 단백발현은 다른 임상지표, 환자의 생존기간과는 관련이 없었다. 결 론 VEGF단백 발현은 두경부 편평세포암 환자에서 나쁜 임상경과를 예측할 수 있게 하는 표지자로서의 역할을 할 수 있다. 또한 본 연구는 두경부 편평세포암에서 별로 연구되지 않은 Apaf-1과 caspase-9의 발현상태를 밝힌 점에서 의의가 있다.

• International Journal of Oral Biology
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• 제39권1호
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• pp.23-33
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• 2014

Several studies have shown that curcumin, which is derived from the rhizomes of turmeric, possesses antimicrobial, antioxidant and anti-inflammatory properties. The antitumor properties of curcumin have also now been demonstrated more recently in different cancers. This study was undertaken to investigate the modulation of cell cycle-related proteins and the mechanisms underlying apoptosis induction by curcumin in the SCC25 human tongue squamous cell carcinoma cell line. Curcumin treatment of the SCC25 cells resulted in a time- and dose-dependent reduction in cell viability and cell growth, and onset of apoptotic cell death. The curcumin-treated SCC25 cells showed several types of apoptotic manifestations, such as nuclear condensation, DNA fragmentation, reduced MMP and proteasome activity, and a decreased DNA content. In addition, the treated SCC25 cells showed a release of cytochrome c into the cytosol, translocation of AIF and DFF40/CAD into the nuclei, a significant shift in the Bax/Bcl-2 ratio, and the activation of caspase-9, caspase-7, caspase-6, caspase-3, PARP, lamin A/C, and DFF45/ICAD. Furthermore, curcumin exposure resulted in a downregulation of G1 cell cycle-related proteins and upregulation of $p27^{KIP1}$. Taken together, our findings demonstrate that curcumin strongly inhibits cell proliferation by modulating the expression of G1 cell cycle-related proteins and inducing apoptosis via proteasomal, mitochondrial, and caspase cascades in SCC25 cells.

• BMB Reports
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• 제48권9호
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• pp.531-536
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• 2015

Gecko proteins have long been used as anti-tumor agents in oriental medicine, without any scientific background. Although anti-tumor effects of Gecko proteins on several cancers were recently reported, their effect on bladder cancer has not been investigated. Thus, we explored the anti-tumor effect of Gecko proteins and its cellular mechanisms in human bladder cancer 5637 cells. Gecko proteins significantly reduced the viability of 5637 cells without any cytotoxic effect on normal cells. These proteins increased the Annexin-V staining and the amount of condensed chromatin, demonstrating that the Gecko proteinsinduced cell death was caused by apoptosis. Gecko proteins suppressed Akt activation, and the overexpression of constitutively active form of myristoylated Akt prevented Gecko proteins-induced death of 5637 cells. Furthermore, Gecko proteins activated caspase 9 and caspase 3/7. Taken together, our data demonstrated that Gecko proteins suppressed the Akt pathway and activated the intrinsic caspase pathway, leading to the apoptosis of bladder cancer cells. [BMB Reports 2015; 48(9): 531-536]

• Molecules and Cells
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• 제20권2호
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• pp.189-195
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• 2005

Ethanol has long been implicated in triggering apoptotic neurodegeneration. We examined the effects of ethanol on the rat brain during synaptogenesis when a spurt in brain growth occurs. This period corresponds to the first 2 postnatal weeks in rats and is very sensitive to ethanol exposure. Ethanol was administered subcutaneously to 7-day- postnatal rat pups by a dosing regimen of 3 g/kg at 0 h and again at 2 h. Blood ethanol levels peaked ($677{\pm}16.4mg/dl$) at 4 h after the first ethanol administration. The cerebral cortexes of the ethanol-treated group showed several typical symptoms of apoptosis such as chromosome condensation and disintegration of cell bodies. Activated caspase-3 positive cells were found in the cortex within 2 h of the first injection, and reached a peak at 12 h. In addition, TUNEL staining revealed DNA fragmentation in the same regions. These results demonstrate that acute ethanol administration causes neuronal cell death via a caspase-3-dependent pathway within 24 h, suggesting that activation of caspase-3 is a marker of the developmental neurotoxicity of ethanol.

• The Korean Journal of Physiology and Pharmacology
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• 제25권4호
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• pp.321-331
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• 2021

Vancomycin, an antibiotic used occasionally as a last line of treatment for methicillin-resistant Staphylococcus aureus, is reportedly associated with nephrotoxicity. This study aimed at evaluating the protective effects of lutein against vancomycin-induced acute renal injury. Peroxisome proliferator-activated receptor gamma (PPARγ) and its associated role in renoprotection by lutein was also examined. Male BALB/c mice were divided into six treatment groups: control with normal saline, lutein (200 mg/kg), vancomycin (250 mg/kg), vancomycin (500 mg/kg), vancomycin (250 mg/kg) with lutein, and vancomycin (500 mg/kg) with lutein groups; they were euthanized after 7 days of treatment. Thereafter, samples of blood, urine, and kidney tissue of the mice were analyzed, followed by the determination of levels of N-acetyl-β-D-glucosaminidase (NAG) in the urine, renal creatine kinase; protein carbonyl, malondialdehyde, and caspase-3 in the kidney; and the expression of PPARγ, nuclear factor erythroid 2-related factor 2 (Nrf2), and nuclear factor-kappaB (NF-κB) in renal tissue. Results showed that the levels of protein carbonyl and malondialdehyde, and the activity of NAG, creatine kinase and caspase-3, were significantly increased in the vancomycin-treatment groups. Moreover, the levels of Nrf2 significantly decreased, while NF-κB expression increased. Lutein ameliorated these effects, and significantly increased PPARγ expression. Furthermore, it attenuated vancomycin-induced histological alterations such as, tissue necrosis and hypertrophy. Therefore, we conclude that lutein protects against vancomycin-induced renal injury by potentially upregulating PPARγ/Nrf2 expression in the renal tissues, and consequently downregulating the pathways: inflammation by NF-κB and apoptosis by caspase-3.

• 한국식품영양과학회지
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• 제45권12호
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• pp.1708-1716
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• 2016

Akt 및 mTOR는 세포 생존에 필수적인 경로로 세포 성장과 증식 등에서 중요한 역할을 하는 것으로 알려져 있다. 본 연구에서는 항암 및 항균 효과가 있는 것으로 알려진 개똥쑥(Artemisia annua L.)에 의한 HepG2 간암세포의 apoptosis 유도 효과를 확인하였다. 본 연구 결과에 의하면 개똥쑥 추출물의 처리 농도가 증가함에 따라 HepG2 세포의 생존율은 억제되었으며, 이는 apoptosis 유도 효과에 의한 것임을 세포의 형태적 변화와 flow cytometry를 통해 확인하였다. 그리고 mitopotential assay와 caspase-3/7 activity assay, western blotting으로 Bcl-2 family 단백질을 확인함으로써 apoptosis 경로 중 내인성 경로(intrinsic pathway)에 의해 apoptosis가 일어남을 알 수 있었다. 이러한 효과는 Akt/mTOR의 활성 저해와 연관이 있었으며 Akt/mTOR의 저해제인 LY294002/rapamycin을 개똥쑥 추출물과 병행처리하였을 경우 개똥쑥 추출물에 의한 apoptosis 효과를 더욱 증대시켰다. 따라서 Akt/mTOR의 저해는 개똥쑥 추출물의 apoptosis 효과를 상승시켰으며 이에 따라 미토콘드리아의 기능 손상과 caspase 활성의 증가를 통해 이루어짐을 확인하였다.

• Clinical and Experimental Pediatrics
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• 제51권10호
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• pp.1112-1117
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• 2008

목 적: 해마 절편 배양에서 산소-포도당 박탈(oxygen-glucose deprivation, OGD)에 의한 세포 사망과 신경 세포 사멸을 propidium iodide(PI) 섭취, Fluoro-Jade(FJ) 염색, TUNEL 염색, caspase-3 면역형광염색 방법으로 관찰하고자 하였다. 방 법: 생후 7일된 Sprague-Dawley 흰쥐의 해마를 MacIlwain chopper로 $350{\mu}m$ 두께의 절편으로 절단하였다. 해마 절편을 6-well plate의 insert 내의 반 유공(sem-porous) 막 위에서 membrane-interface technique으로 10일 동안 배양하였다. 배양된 해마 절편에 산소-포도당 박탈을 60분 동안 가한 후 재산소-재관류하에 기초 배양액에서 48시간 배양하였다. 재산소-재관류 동안 PI 섭취 형광 정도를 시간에 따라 형광 현미경으로 관찰하고 세포사망 백분율(percent cell death)을 측정하였다. 산소-포도당 박탈 직전과 24 시간 후에 해마 절편을 $15{\mu}m$ 두께로 냉동 절단 후 FJ 염색, TUNEL 염색, caspase-3 면역형광염색을 시행하여 세포 사망을 관찰하였다. 결과: OGD 후 PI 섭취 는 해마 절편의 CA1과 DG에 한정되어있었다. OGD 후 재산소-재관류 동안 6시간에서 48시간까지 PI 섭취 형광 강도는 시간이 증가함에 따라 증가하였다. 세포 사망 백분율은 CA1과 DG에서 모두 OGD 후 재산소-재관류 시간이 증가함에 따라 의미 있게 증가하였다(P<0.05). OGD 후 24시간에 세포 변성을 의미하는 많은 FJ 염색 양성 신경 세포 들이 CA1과 DG에서 관찰되었다. 고배율 confocal laser 현미경으로 관찰한 CA1에서의 신경 세포들 중 일부는 명확한 핵과 돌기를 가지고 있는 것을 보여 주었으며, 다른 신경 세포들은 핵의 분절화, 돌기의 손실 등을 보여 주었다. TUNEL 염색과 caspase-3 염색은 OGD 후 24시간에 CA1과 DA에서 TUNEL 양성 발현을 증가시키고 caspase-3 발현을 증가시켰다. 결 론: 해마 절편 배양에서 산소-포도당 박탈 에 의한 다수의 세포 사망을 관찰할 수 있었다. 사망한 세포 들은 주로 신경 세포의 caspase-3 활성화에 의해 매개된 사멸을 보였다.

• Asian Pacific Journal of Cancer Prevention
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• 제14권2호
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• pp.891-894
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• 2013

The present study was performed to assess the activity of the botulinum toxin A on breast cancer cells. The T47D cell line was exposed to diverse concentrations of the botulinum toxin A and cell viability and apoptosis were estimated using MTT and propidium iodine/annexin V methods, respectively. Botulinum toxin A exerted greater cytotoxic activity in T47D cells in comparison with MCF10A normal cells; this appeared to be via apoptotic processes caspase-3 and -7. In conclusion, botulinum toxin A induces caspase-3 and -7 dependent apoptotic processes in the T47D breast cancer cell line.