To obtain further information concerning the nature myofibrillar proteins in a food system, an investigation has been conducted to compare the change in the biochemical property of the myofibril with the changes in the morphological structure of the myofibril. When myofibrils were prepared with 0.16 M KCl-0.04 M Tris-HCl, the band pattern was clear and distinct. There was a uniform thickening of A-band, a sharp appearence of Z-lines and a wide I-band. The band pattern of myofibrils was changed as the composition of extraction solution was changed. Also the ATPase activity of myofibril changed as the length of sarcomere changed. When myofibrils were treated with a low concentration of trypsin, myofibrils turned in the contracted state. With the progress of prolonged trypsin treatment, most of myofibrils exhibited a pattern of alternating light and dark bands, supercontracted pattern. Although myofibrils exhibited a supercontracted band pattern, the ATPase activity of myofibril continued to increase with the progress of trypsin treatment. An assumption was made that tropomyosin may be located in Z-line and that troponin-tropomyosin complex can inhibit the ATPase activity of myofibrils through the structural alternation of myofibril.
Objectives : The effects of aqueous extracts of Cortex ulmi pumilae (a traditional medicine for cancer treatment in oriental medicine) on the induction of apoptotic cell death were investigated in human liver origm hepatoma cell lines, HepG2. Methods : The death of HepG2 cells was markedly induced by the addition of extracts of Cortex ulmi pumilae in a dose-dependent manner. The apoptotic characteristic ladder pattern of DNA strand break was not observed in cell death of HepG2. In addition, it was not shown nucleus chromatin condensation and fragmentation under hoechst staining. However, by the using annexin V staining assay, externalizations of phosphatidylserine in HepG2 cell which were treated with Cortex ulmi pumilae extracts were detected in the early time (at 9 hr after extract treatment). Furthermore, LDH release was not detected in this early stage. Therefore, Cortex ulmi pumilae extracts-induced cell death of HepG2 cells is mediated by apoptotic death signal processes. Result : The activity of caspase 3-like proteases remained in a basal level in HepG2 cells which treated with the extract of Cordyceps sinensis. However, it was markedly increased in HepG2 cells which treated with two extracts of Cortex ulmi pumilae (C.U.P.-C, C.U.P.-K) which were differently extracted (respectively, 2.3 and 3.3 fold). On a while, the phosphotransferase activities of JNK1 was markedly induced in HepG2 cells which were treated with two extracts of Cortex ulmi pumilae. On the contrary, the activation of transcriptional activator, activating protein1(AP-1) and NF-kB were severely decreased by these two extracts of Cortex ulmi pumilae (C.U.P.-C, C.U.P.-K). In addition, antioxidants (GSH and NAC) and intracellular $Ca2^+$ level regulator (Bapta/AM and Thapsigargin) did not affect Cortex ulmi pumilae extracts-induced apoptotic death of HepG2 cells. Conclusions : In conclusion, our results suggest that two extracts of Cortex ulmi pumilae (C.U.P.-C, C.U.P.-K) induces the apoptotic death of human liver origin hepatoma HepG2 cells via activation of caspase 3-like proteases as well as JNK1, and inhibition of transcriptional activators, AP-1 and $NK-{\kappa}B$.
Objective : This study was designed to evaluate the synergistic effect on cytotoxicity of combination with adriamycin and Palginhonhapwhajucwhan, a traditional prescription for cancer treatment in oriental medicine, in Chang, HL-60, Hep-3B and Alexander cells. Methods : We observed cell viability in Chang, HL-60, Hep-3B, and Alexander cells by crystal violet staining. Those cells were treated with various concentrations of adriamycin alone, Palginhonhapwhajucwhan alone and combination of two medications for 10 hr. On condition of $0.5{\mu}l/ml$ adriamycin alone, $15.6{\mu}l/ml$ Paljintanghapwhajucwhan alone and combination of two medications, at first, we observed colony forming of Chang and HL-60 cells. Second, we observed DNA fragmentation by agarose electrophoresis in Chang, HL-60, Hep-38 and Alexander cells. Third, we measured the catalytic activation of caspase-1, 2, 3, 6, 8, and 9 protease in Chang cells and caspase-3 protease in Chang, HL-60, Hep-3B and Alexander cells by using fluorogenic substrate. Finally, we isolated mRNA of Fas in Chang, HL-60, Hep-38 and Alexander cells and observed that Fas gene was amplified by RT-PCR Results : 1. The combination of adriamycin and Palginhonhapwhajucwhan synergistically augmented the cytotoxicity of Chang and HL-60 cells whereas did not in Hep-38 and Alexander cells. 2. Cotreatment of two drugs also markedly inhibited the colony forming ability both in Chang and HL-60 cells. 3. The cytotoxicity of these medicatons was revealed as apoptosis characterized by high molecular wight DNA fragmentaton. 4. The apoptotic cytotoxicity was mediated by activation of caspase-3 protease in Chang cells. 5. Synergistic increase in apoptotic cytotoxicity by combination of two medications was dependent on the expression of Fas in cancer cells. Conclusions : Combination of adriamycin and Palginhonhapwhajucwhan significantly augmented apoptotic cytotoxicity of Fas-positive cells such as Chang and HL-60 cells via acticaton of apoptosis signaling pathway.
Jeon In Cheol;Jeong Jae Eun;Son In Hwan;Lee Ju Seok;Jeong Seung Won;Jang Jae Ho;Lee Seon U;Lee In;Moon Byun Soon
Journal of Physiology & Pathology in Korean Medicine
/
v.18
no.6
/
pp.1652-1660
/
2004
Apoptosis of vascular smooth muscle cells(VSMCs) is essential in atherogenesis, being a factor that modulates its early progression rather than a terminal event in the course of the disease. Various stimuli, including oxide lipoproteins, altered hemodynamic stress and free radical, can induced VSMCs apoptosis in vitro. The protective effects of Sophorae Radix (SR) on apoptotic cell death induced by H₂O₂ were investigated in VSMCs. The viability of VSMCs was markedly decreased by H₂O₂. Sophorae Radix protected the H202-induced apoptotic death of VSMCs, which was characterized as nuclear fragmentation and increase of sub-G0/G1 fraction .. Sophorae Radix decreased the activation of caspase-3 like protease induced by H₂O₂ and recovered control level from H202-induced PARP, Bak, Bcl-XL and mitochondrial membrane potential. These results suggest that Sophorae Radix protected VSMCs apoptotic death induced by H₂O₂ via inactivation of caspase-3 and modulation of mitochondrial function. Also, the expression profile of proteins by using two-dimensional (2-D) gel electrophoresis was screened. Future investigations will need to explore the use of an anti atherosclerotic therapy of Sophorae Radix, which relies on inhibition of the proapoptotic activation of the vascular smooth muscle cells.
Park Young Jun;Jung Woo Cheal;Jeong Dae Young;Lee Yong Un;Lee In;Lee Key Sang;Jeon Byung Hun;Sung Kang Keyng;Moon Byung Soon
Journal of Physiology & Pathology in Korean Medicine
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v.17
no.6
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pp.1383-1392
/
2003
Apoptosis is a morphologically and biochemically district form of cell death that occurs in many different cell types in a wide variety of organisms. Albizzia 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 of A. 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 of A. julibrissin to induce apoptosis was associated with proteolytic cleavage of specific target proteins such as poly (ADP-ribose)polymerase (PARP) and beta-catenin proteins suggesting the possible involvement of caspases. Our result showed 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. The purpose of the present study is also to investigate the Effect of A. julibrissin on cell cycle progression. Our results showed that G1 checkpoint related gene products (cyclin D1, cyclin dependent kinase 4, retinoblastoma, E2F1) were decreased in their protein levels in a dose-dependent manners after treatment of the extract. These results indicate that the increase of apoptotic cell death by A. julibrissin may be due to the inhibition of cell cycle progression in wild type p53-lacking Jurkat cells.
Jun Soo Young;Shin Dong Hoon;Son Chang Woo;Shin Heung Mook
Journal of Physiology & Pathology in Korean Medicine
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v.18
no.4
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pp.1055-1060
/
2004
Apoptosis is the ability of cells to self-destruct by the activation of an intrinsic cellular suicide program when the cells are no longer needed or when they are seriously damaged. Morphologically, apoptosis is characterized by the appearance of membrane blebbing, cell shrinkage, chromatin condensation, DNA cleavage, and the fragmentation of the cell membrane-bound apoptotic bodies. Siegesbeckia glabrescens Makino (Siegesbeckiae Herba, SG) has been widely used as treatments for arthritis, and fever, as well as detoxification properties. The present studies were undertaken to evaluate if SG has an anti-apoptotic property. Cell viability was measured by XTT and tryphan blue stain. Morphological characteristic of human aortic smooth muscle cells(HASMC) were visualized with a phase-contrast microscope. SG significantly reduced HASMC, but not human umbilical vein endothelial cell(HUVEC), viability in a dose-dependent manner. Confluent untreated cells at 24hrs showed normal morphology, flat with a uniform polygonal shape. SG-treated cells (0.5㎎/㎖) at 24hrs showed apoptotic morphology. Cells became irregular with elongated lamellipodia, and exhibited condensed chromatin in nuclei with occasional endoucleation. There was an increase in the number of apoptotic cells rounding-up and being detached from the substrate. TUNEL staining of SG-treated cells showed dark-brown stains in nuclei and cytosol. Caspases are central components of the machinery responsible for apoptosis and are generally divided into two categories; the initiator caspases, which include caspases-2,-8,-9, and -10, and the effector caspases, which include caspases-3,-6, and -7. SG decreased anti-caspase-3 protein expression, which means activation of caspases-3 activity. It has been reported that there is a link between NO formation and apoptosis. NO production was accelerated by SG treatment in HASMC. L-NNA, NOS inhibitor, inhibited SG-induced apoptosis. These results, therefore, indicated that both caspases-3 and NO production are involved in apoptosis in smooth muscle cells. According to these results, SG may have a potential effect in the treatment of hypertensive atherosclerosis.
Sung, Nak Yoon;Kim, Seung Cheol;Kim, Yun Hwan;Kim, Gihyeon;Lee, Yunmi;Sung, Gi-Ho;Kim, Ji Hye;Yang, Woo Seok;Kim, Mi Seon;Baek, Kwang-Soo;Kim, Jong-Hoon;Cho, Jae Youl
Biomolecules & Therapeutics
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v.24
no.4
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pp.402-409
/
2016
It has been found that 4-isopropyl-2,6-bis(1-phenylethyl)phenol (KTH-13), a novel compound isolated from Cordyceps bassiana, is able to suppress tumor cell proliferation by inducing apoptosis. To mass-produce this compound, we established a total synthesis method. Using those conditions, we further synthesized various analogs with structural similarity to KTH-13. In this study, we aimed to test their anti-cancer activity by measuring anti-proliferative and pro-apoptotic activities. Of 8 compounds tested, 4-methyl-2,6-bis(1-phenylethyl)phenol (KTH-13-Me) exhibited the strongest anti-proliferative activity toward MDA-MB 231 cells. KTH-13-Me also similarly suppressed the survival of various cancer cell lines, including C6 glioma, HCT-15, and LoVo cells. Treatment of KTH-13-Me induced several apoptotic signs in C6 glioma cells, such as morphological changes, induction of apoptotic bodies, and nuclear fragmentation and chromatin condensation. Concordantly, early-apoptotic cells were also identified by staining with FITC-Annexin V/PI. Moreover, KTH-13-Me highly enhanced the activation of caspase-3 and caspase-9, and decreased the protein level of Bcl-2. In addition, the phosphorylation levels of Src and STAT3 were diminished in KTH-13-Me-treated C6 cells. Therefore, these results suggest that KTH-13-Me can be developed as a novel anti-cancer drug capable of blocking proliferation, inducing apoptosis, and blocking cell survival signaling in cancer cells.
The present study was performed to investigate the effect of Yangkyuksanhoa-tang on the morphological changes of the basilar artery after experimentally induced subarachnoid hemonrrhages(SAH). Yangkyuksanhoa-tang has been used freguently for cerebrovascular accident Sprague Dawley rats weighing between 350-400 g were used. The 6 normal rats and 24 SAH elicited rats were used, The SAH induced by injection of the fresh autologus heart blood (0.3-0.4 ml) into the cisterna magna through the posterior atlanta-occipital membrane, Sample group was given 3.3 ml/kg/day of Yangkyuksanhoa-tang extracts for 2 days after SAH. The experimental animals were killed at 48hrs after SAH. The morphological changes of the arterial walls were examined by light and electron microscopy. Following are the obtained results: 1. In SAH elicited rats, the size of the lumen in basilar artery was diminished by about 45% and the thickness of arterial wall was increased by about 82%. In SAH elicited rats with Yangkyuksanhoa-tang treatment, the size of the lumen in basilar artery was merely diminished by about 18% and the thickness of arterial wall was merely increased by about 19%. 2. In light microscopic examination, the endothelium was swollen into a cuboid shape and the layer of smooth muscle was increased in the basilar artery of SAH elicited rats. In SAH elicited rats with Yangkyuksanhoa-tang treatment, the size of the lumen in basilar artery was enlarged and the thickness was decreased than in SAH elicited rats. The endothelium was flattened into a squamous shape and the layer of smooth muscle was decreased more than in SAH elicited rats. 3. In electron microscopic examination, the endothelial cells with fragmentation nuclei were changed into a cuboid shape and the internal elastic lamina were folded at the basilar artery of SAH elicited rat. The nuclei of smooth muscle cells were changed into a round or crumpled shape. The length of smooth muscle was shorten and thickness was increased. But all kinds of morphologic changes were diminished in SAH elicited rats with Yangkyuksanhoa-tang treatment. Conclusion : Yangkyuksanhoa-tang extracts were effective to treat cerebral vasospasm after experimentally induced subarachnoid hemorrhage in rats.
Journal of the Korea Academia-Industrial cooperation Society
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v.11
no.12
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pp.4945-4953
/
2010
The purpose of this study was to analyze the differences in perception levels of goals and concepts of physical education by the body image and gender of middle school students. Among 461 middle students randomly selected by a multistaged cluster sampling technique, 433 students(boys 267, girls 166) filled out questionnaires consisted of Figure Rating Scale and modified ones. To know the differences in the perception levels of goal and concept of physical education by sex, t-test was utilized. In order to find out the differences in the perception levels of goal and concept of physical education by body image, ANOVA was performed using the SPSS Windows 12.0. The results of this study revealed that boys had higher level of goal and concept of physical education than girls. Second, middle school students classified into normal body image was higher level of goal and concept of physical education that other body images such as very thin, thin, very obese, and obese ones. The results of this study indicated that curriculum of physical education focused on fragmentation of health, psychomotor, cognitive, affective, leisure, holistic human should make girls understand thoroughly and deeply goals and concepts of physical education and ideal body image.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.30
no.6
/
pp.509-515
/
2004
Photodynamic therapy(PDT) has advanced to clinical trials for the treatment of a variety of solid tumors and presents an alternative treatment option for tumors resistant to chemo-and/or radio-therapy. PDT is based on the combination of laser light of appropriate wavelength and energy to activate a systemically or locally applied photosensitizer that concentrates preferentially in malignant tissues. In this study, phototoxicity of laser EIT 21 was analysed in human osteosarcoma cell(HOS) and the second objective of this study was to determine the ability of laser EIT 21 to induce apoptosis. This study demonstrated that laser EIT 21 had a phototoxicity to HOS cells. In order to examinate whether cell death was induced by necrosis or apoptosis, variety of techniques which assess apoptosis were used. TUNEL assay showed only a few the positive reaction on condensed nuclei. It is hard to find condensed or fragmented nuclei on HOS cells irradiated with laser EIT 21 in Hemastat and AO/EB stain. By DNA electrophoresis, cells also did not show DNA degradation characteristic of apoptosis with a ladder pattern of DNA fragments. Apoptosis-related factors were analyzed by western blotting. The expression of p53 was constant and cells irradiated with laser did not show the caspase-3 and PARP degradation, therefore we suggest that p53 and caspase-3 are not involved in laserinduced cell death.
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