• Nutrition Research and Practice
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• 제9권6호
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• pp.586-591
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• 2015

BACKGROUND/OBJECTIVES: Reactive oxygen species (ROS) formation is closely related to miconazole-induced heart dysfunction. Although rhamnetin has antioxidant effects, it remained unknown whether it can protect against miconazole-induced cardiomyocyte apoptosis. Thus, we investigated the effects of rhamnetin on miconazole-stimulated H9c2 cell apoptosis. MATERIALS/METHODS: Cell morphology was observed by inverted microscope and cell viability was determined using a WelCount$^{TM}$ cell proliferation assay kit. Miconazole-induced ROS production was evaluated by fluorescence-activated cell sorting with 6-carboxy-2',7'-dichlorofluoroscein diacetate ($H_2DCF$-DA) stain. Immunoblot analysis was used to determine apurinic/apyrimidinic endonuclease 1 (APE/Ref-1) and cleaved cysteine-aspartic protease (caspase) 3 expression. NADPH oxidase levels were measured using real-time polymerase chain reaction. RESULTS: Miconazole (3 and $10{\mu}M$) induced abnormal morphological changes and cell death in H9c2 cells. Rhamnetin enhanced the viability of miconazole ($3{\mu}M$)-treated cells in a dose-dependent manner. Rhamnetin (1 and $3{\mu}M$) treatment downregulated cleaved caspase 3 and upregulated APE/Ref-1 expression in miconazole-stimulated cells. Additionally, rhamnetin significantly reduced ROS generation. CONCLUSIONS: Our data suggest that rhamnetin may have cytoprotective effects in miconazole-stimulated H9c2 cardiomyocytes via ROS inhibition. This effect most likely occurs through the upregulation of APE/Ref-1 and attenuation of hydrogen peroxide levels.

• Fisheries and Aquatic Sciences
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• 제18권1호
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• pp.81-88
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• 2015

We investigated factors affecting primary cultures of Pacific abalone Haliotis discus hannai ovary-dissociated cells to identify general aspects of their early-phase culture. Ninety-seven cell populations derived from 30 individuals were cultured in different media with varying compositions of medium supplements, and initial attachment, subculture, and survival for ${\geq}10$ weeks were assessed according to medium composition and individual. We also examined the time required for subculture and the rate of cell death according to both culturing period and passage number within 10 weeks. A lack of fetal bovine serum (FBS) and hemolymph significantly inhibited the growth of cultured cells, while we detected no significant effect of medium composition on initial cell attachment. Through data reallocation, with the omission of data from cell populations cultured in FBS-free and hemolymph-free media, we showed that growth inhibition was also affected by individual differences among the abalones used. During the culture, we observed four different types of cell morphology. Moreover, considerable time was required for subculture-18.4 and 19.5 days for first and second subcultures, respectively-and cell death did not occur within 30 days or for passage 0. Our results will provide valuable information for developing universal cell culturing guidelines in abalone species and suggest the feasibility of culturing abalone ovary-dissociated cells.

• 약학회지
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• 제45권6호
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• pp.730-738
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• 2001

Apoptosis is a morphologically and biochemically distinct form of cell death that occurs in many different cell types in a wide variety of organisms. Ajbizzia julibrissin belonging the family Leguminosae has been used for the treatment of contusion, sore throat, amnesia, and insomnia in oriental traditional medicine. This study investigates whether the water extract off julibrissin induce apoptotic cell death in Jurkat T-acute lymphoblastic leukemia (ALL) cells. Jurkat cells were increased inhibitions of cell viability in a concentration-dependent manner by A julibrissin. This herbal medicine also caused apoptosis as measured by cell morphology and DNA fragmentation. The capability oft julibrissin to induce apoptosis was associated with proteolytic cleavage of specific target protein such as poly (ADP-ribose) polymerase (PARP) protein suggesting the possible involvement of caspases. Our result skewed that Bcl-2 and Bax protein levels were not changed in all A julibrissin-treated groups compared to control group. These results suggest that A julibrissin-mediated apoptosis is independent with Bcl-2 related signaling pathway in this cells.

• 대한본초학회지
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• 제25권2호
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• pp.107-116
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• 2010

Objectives : This study purposed to research the anti-cancer effects of Andrographitis Herba. Methods : By measuring the cell proliferation, apoptosis, morphology and cytokine level from the extracts, the influence on a A549 cell was compared. Results : The Andrographitis Herba decoction extract according to the concentration inhibited the proliferation and increased the apoptosis of the A549 cell. Among the various fraction extracts of the Andrographitis Herba decoction, EtOEt showed the greatest increase of the apoptosis of the A549 cell. The Andrographitis Herba decoction extract according to the concentration decreased the secretion of the TGF-$\beta$ in the A549 cell, and increased the secretion of the TNF-$\alpha$ and the IFN-$\gamma$ presenting cell population. Conclusion : It is considered that the total extract and various fraction extracts of Andrographitis Herba decoction inhibit the proliferation of A549 cells.

• Journal of Microbiology and Biotechnology
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• 제18권7호
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• pp.1252-1256
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• 2008

Cell recycled culture of succinic acid-producing Anaerobiospirillum succiniciproducens was anaerobically carried out using an internal membrane filter module in order to examine the physiological response of A. succiniciproducens to a high-cell-density environment. The optimal growth of A. succiniciproducens and its enhanced succinic acid productivity were observed under $CO_2$-rich conditions, established by adding $NaHCO_3$ and $Na_2CO_3$, in the cell recycled system. A. succiniciproducens grew up to 6.50 g-DCW/l, the highest cell concentration obtained so far, in cell recycled cultures. The cells did not change their morphology, which is known to be easily changed in unfavorable or stress environments. The maximum productivity of succinic acid was about 3.3 g/l/h, which is 3.3 times higher than those obtained in batch cultures. These results can serve as a guide for designing highly efficient cell recycled systems for succinic acid at a commercial level.

• The Journal of Korean Physical Therapy
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• 제21권3호
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• pp.87-93
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• 2009

Purpose: TThis study examined the effect of repetitive magnetic stimulation (RMS) on the viability and proliferative response of human adipose tissue-derived stromal cells (hATSCs) in vitro. Methods: The hATSCs were cultured primarily from human adipose tissue harvested by liposuction and incubated in a $37^{\circ}C$ plastic chamber. The cells were exposed to a repetitive magnetic field using a customized magnetic stimulator (Biocon-5000, Mcube Technology). The RMS parameters were set as follows: repetition rate=10Hz, 25Hz (stimulus intensity 100%= 0.1 Tesla, at 4cm from the coil), stimulated time= 1, 5, and 20 minutes. Twenty four hours after one application of RMS, the hATSCs were compared with the sham stimulation, which were kept under the same conditions without the application of RMS. The cells were observed by optical microscopy to determine the morphology and assessed by trypan blue staining for cell proliferation. The apoptosis and viability of the hATSCs were also analyzed by fluorescence-activated cell sorting (FACS) analysis of Annexin V and MTT assay. Results: After RMS, the morphology of the hATSCs was not changed and the apoptosis of hATSCs were not increased compared to the sham stimulation. The viability of the cells was similar to the cells given the sham stimulation. Interestingly, the level of hATSC proliferation was significantly higher in all RMS groups. Conclusion: The application of RMS may not cause a change in morphology and viability of hATSCs but can increase the level of cell proliferation in vitro. RMS might be useful as an adjuvant tool in combination with stem cell therapy without adverse effects.

• 한국가금학회지
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• 제32권1호
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• pp.49-54
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• 2005

효모(Saccharomyces cerevisiae, SC)의 구성성분인 yeast cell-extract(YE)과 yeast cell-wall(CW)이 육계의 생산성, 장내융모 발달 및 혈청 콜레스테롤에 어떠한 영향을 미치는지 알아보고자 사양실험을 실시하였다. 육용 수평아리(Ross) 240수를 4처리 6반복, 반복당 10수를 공시하였다. 옥수수 및 대두박 위주 기본사료에 SC, YE 그리고 CW를 각각 0.5, 0.25 그리고 $0.25\%$로 첨가한 실험사료를 제조하여 육계에 5간 급여하였다. 효모의 급여는 육계의 생산성에 어떠한 영향을 미치지 못하였으며, villus height, crypt depth 및 villus:crypt ratio 역시 처리간 유의성은 발견되지 않았다. SC 급여 받은 육계의 혈청 콜레스테롤은 대조구에 비하여 $19.7\%$ 감소(P<0.05)를 나타내었다. 특히, 효모성분 중 YE가 혈청 콜레스테롤을 유의적으로 낮추었다(P<0.05). 결론적으로 사료내 효모의 첨가는 육계의 혈청 콜레스테롤을 유의적으로 감소시켰으며(P<0.05), 콜레스테롤을 감소시키는 성분은 효모의 세포내용물에 포함되어 있는 것으로 본 연구결과 알 수 있었다.

• 한국패류학회지
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• 제26권3호
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• pp.235-244
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• 2010

Ultrastructural characteristics of the testis and spermatogenesis of Crassostrea gigas were investigated by Transmission and Scanning Electron microscope observations. The testis is a diffuse organ consisting of branching acini containing differentiating germ cells in a variety of stages. The acinus is surrounded by an intermitent layer of myoepithelial cells andis divided into subcompartments that are partially separated by pleomorphic accessory cells which remain in close contact with germ cells until late stages of development. these accessory cells contain a large quantity of glycogen particles and lipid droplets in the cytoplasm. Therefore, it is assumed that they are involved in the supplying of the nutrients for germ cell development, while any phenomena associated with phagocytosis of undischarged, residual sperms by lysosomes could be find in the cytoplasm of the accessory cells. The morphology of the spermatozoon has a primitive type and is similar to those of other bivalves. Mature spermatozoa consist of broad, cap-shaped acrosomal vesicle, subacrosomal material (containing axial rod embedded in a granular matrix), a oval nucleus showing deeply invaginated anteriorly, two triplet substructure centrioles surrounded by four spherical mitochondria, and satelite fibres appear to the distal centriole and plasma membrane. Spermatozoa of C. gigas resemble to those of other investigated ostreids. In particular, the anterior region of the acrosomal vesicle is transversely banded. It is assumed that differences in this acrosomal substructure are associated with the inability of fertilization between the genus Crassostrea and other genus species in Ostreidae. Therefore, we can use sperm morphology in the resolution of taxonomic relationships within the Ostreidea. The spermatozoon is approximately $42-47{\mu}m$ in length including an oval sperm nucleus (about $0.91{\mu}m$ in length), an acrosome (about $0.42{\mu}m$ in length) and tail flagellum ($40-45{\mu}m$). The axoneme of the sperm tail flagellum consists of nine pairs of microtubules at the periphery and a pair at the center. The axoneme of the sperm tail shows a 9 + 2 structure. These morphological charateristics of acrosomal vesicle belong to the family Ostreidae in the subclass Pteriomorphia.

• 동의생리병리학회지
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• 제23권5호
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• pp.1139-1144
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• 2009

This study was purposed to research the anti-cancer effects of Bistortae Rhizoma. A total extract of Bistortae Rhizoma decoction was prepared. By measuring the cell proliferation, apoptosis, morphology and cytokine level from the extracts, the influence on HepG2 cell, SNU-1 cell and A549 cell was compared. The Bistortae Rhizoma decoction extract did not control HepG2 cell proliferation but controlled SNU-1 cell and A549 cell proliferation. In particular, the inhibitory effect on SNU-1 cell proliferation was highest. The Bistortae Rhizoma decoction extract showed to increase the apoptosis of the HepG2 ceil, SNU-1 cell and A549 cell in a dose-dependent manner. In particular, the promotion effect of the apoptosis was highest in SNU-1 cell. Among the various fraction extracts of the Bistortae Rhizoma decoction, n-BuOH extraction showed the greatest increase of the apoptosis of the HepG2 cell. The Bistortae Rhizoma decoction extract decreased dose-dependently the secretion of the TGF-$\beta$ in the HepG2 cell, SNU-1 cell and A549 cell and increased the secretion of the TNF-$\alpha$ and the IFN-$\gamma$. These results suggest that the total extract of Bistortae Rhizoma decoction has anti-cancer effect against SNU-1 cell and A549 cell.

• Journal of Periodontal and Implant Science
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• 제23권1호
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• pp.77-96
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• 1993

정상치아와 이환치아 및 구연산에 의하여 탈회한 치근면에대한 치주인대세포의 부착상태를 비교관찰 하기 위하여 정상치아와 이환치아를 취하여 정상군, 이환치근의 치근면활택술군 및 구연산처리군으로 분류하고 시험관적 실험을 통하여 관찰하였다. 세포부착실험전 각 군의 치근면의 형태를 주사전자현미경을 이용하여 관찰하였으며, 사람의 정상소구치에서부터 얻어진 치주인대세포를 배야하여 각 군의 절편을 함유한 조직배양기에 ml당 $4.5{\times}\;10^4$개의 세포를 가진 배양액 1ml씩을 넣고 동일조건하에서 30분, 1시간, 2시간, 6시간, 12시간 및 24시간 동안 배양한 후 세포의 부착이 일어난 후 상태를 주사전자현미경을 이용하여 관찰하였고, 세포증식정도를 알아보기 위하여 각 절편에 $5{\times}10^4$개의 세포가 함유된 배양액 1ml씩을 넣고 6시간동안 배양 후, 새 배양기에 옮기고 24, 48, 72시간동안 배양하여 trypsin 처리로 세포를 분리시킨 후 광학위상차 현미경을 이용, 치근단위면적당 증식된 세포수를 측정하여 다음과 같은 결과를 얻었다. 세포부착실험전 각 군의 치근면 형태를 관찰하였을 때 구연산처리군에서는 교원섬유의 노출부위라고 여겨지는 미세한 돌출구조들을 관찰할 수 있었으며 상아세관의 노출부위라고 여겨지는 함몰양상이 치근활택술군에 비하여 현저하였고 정상치아군에서는 이러한 양상을 관찰할 수 없었다. 실험 각 군 및 대조군 사이에서 세포들의 형태적인 차이점은 분명하지 않았으나 구연산처리군에서 다른 군들보다 초기의 세포부착양상이 다소 빠르게 진행 됨이 관찰되었다. 세포배양 개시 후 6시간이 경과 한 후에는 모든 군에서 공히 세포의 형태가 초기의 구형의 형태에서 편평한 세포의 형태로 변화되기 시작함이 관찰 되었고 24시간 이후에는 모든 군에서 세포들이 치근면에 납작하게 부착되고 일반적인 섬 유아세포의 형태로 변화되어 셰포의 전개가 완성된 양상이 보였다. 세포증식율의 실험에서는 24시간 후 증식된 세포 수는 치근면활택술군에서 가장높게 나타났으며 정상군에서 가장 낮은 수치를 보였으며 48시간 및 72시간후 측정에서는 구연산처리군이 다른 군들에 비해 더 많은 수의 세포증식을 관찰할 수 있었으며 각 군간의 차이는 통계학적으로 유의하였다.