Objectives : In this study, it was investigated the anti-inflammatory, anticancer, and antioxidant effects of Jayangdaebo-tang (JDT) consisting of twelve herbs before and after fermentation. Methods : JDT extract was fermented using the Lactoplantibacillus plantanum (JDT-L), Bacillus subtilis (JDT-B), and L. plantanum plus B. subtilis (JDT-L+B). The effects of each extract were measured in LPS-stimulated RAW264.7 cells, MCF-7 breast cancer and A549 lung cancer cells, and H2O2-stimulated HepG2 cells. Results : The extracts of JDT-L, JDT-B and JDT-L+B at 1 ㎎/㎖ decreased significantly the levels of nitric oxide (NO) in LPS-treated RAW264.7 cells and also inhibited the expression of iNOS and COX-2, and the phosphorylation of ERK and NF-κB. The JDT-L+B extract decreased significantly the expression of apoptotic proteins, Bax, cleaved caspase-3, and PARP in MCF-7 and A549 cancer cells. The JDT-L, JDT-B and JDT-L+B extracts increased significantly the cell viability in H2O2-stimulated HepG2 cells and the JDT-L+B extract decreased significantly the expression of SOD, catalase, HO-1, and NRF-2. Among fermented JDT extracts, JDT-L+B was the best effective on response of macrophage inflammation, cancer cell apoptosis, and liver cell damage. Conclusions : Our results were suggested that the fermentation can be used as a useful way to enhance the biological activity of JDT.
Kim, Sun-Mo;Yun, Hyun-Jeong;Yi, Hyo-Seung;Won, Chan-Wook;Kim, Jai-Eun;Park, Sun-Dong
The Korea Journal of Herbology
/
v.24
no.4
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pp.77-86
/
2009
Objectives : The proliferation and migration of human aortic smooth muscle cells (HASMC) in response to activation by various stimuli plays a critical role in the initiation and development of atherosclerosis. This study was conducted to examine the effects of Nelumbo nucifera leaves (NNL) on the proliferation and migration of HASMC. Additionally, the mechanisms involved in any observed effects were also evaluated. Methods : Apoptotic cells were measured by staining with FITC-labeled annexin V, followed by flow cytometric analysis. The expression level of apoptosis related proteins was confirmed by western blot. And MMP-9 activity was measured by gelatin zymography and MMP-9 expression was measured by ELISA Results : NNL completely inhibited the proliferation of HASMC via induction of the expression of apoptotic proteins including annexin V, cleaved poly ADP-ribose polymerase (PARP), and caspase-3 and -8. NNL treatment resulted in the release of cytochrome c into cytosol, a loss of mitochondrial membrane potential, a decrease in Bcl-2 and Bcl-xL and an increase in Bax expression. NNL also blocked HASMC migration via suppression of MMP-9. Conclusions : Taken together, these results indicate that NNL has the potential for use as an anti-artherosclerosis agent.
Epidemiology studies have reported a reduced incidence of colon cancer among populations that consume a large quantity of ${\omega}3-polyunsaturated$ fatty acids (${\omega}3-PUFAs$) of marine origin. Herein, we demonstrated a mechanism of anticancer action of ${\omega}3-PUFAs$, showing that they suppressed invasion and tumorigenicity in colon cancer cells. Docosahexaenoic acids (DHA) inhibited the cell growth of HT29 cells. This action likely involved apoptosis, given that the DHA treatment increased the cleaved form of PARP and sub G1 cells. Moreover, the invasiveness of HT29 cells was inhibited following DHA treatment, whereas arachidonic acid (AA) had no effect. The levels of Matrix-metalloproteinase-9 (MMP-9) and MMP-2 mRNA decreased after DHA pretreatment. DHA treatment inhibited MMP-9 and MMP-2 promoter activities and reduced VEGF promoter activity. DHA pretreatment also inhibited the activities of prostaglandin-2 (PGE2)-induced MMPs and the VEGF promoter. Cyclooxygenase-2 (COX-2) overexpression increased the activity of MMPs and that of the Vascular endotherial growth factor (VEGF) promoter in HT29 cells, and DHA inhibited NF-kB and COX-2 promoter reporter activities. As shown by in vivo experiments, when mouse colon cancer cells (MCA38) were implanted into Fat-1 and wild-type mice, both the tumoral size and volume were dramatically inhibited in Fat-1 transgenic mice. Furthermore, TUNEL-positive cells increased in tumors from Fat-1 mice compared with wild mice. In immunohistochemistry, the intensity of CD31 in Fat-1 tumors was weaker. These findings suggest that ${\omega}3-PUFAs$ may inhibit tumorigenicity and angiogenesis as well as cancer cell invasion by suppression of COX-2, MMPs and VEGF via the reduction of NF-kB in colon cancer.
Jung, Gi Hwan;Kim, Sung Hyun;Woo, Joong Seok;Yoo, Eun Seon;Lee, Jae Han;Han, So Hee;Jung, Soo Hyun;Kim, Sae Man;Kim, Eun Gee;Shin, Man Jae;Cho, Ho Min;Jung, Ji Youn
Journal of Life Science
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v.31
no.9
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pp.827-833
/
2021
Dendropanax morbiferus leaves (DPL) has been used as a medicine since ancient times in various diseases such as inflammation, diabetes, and cancer. In particular, it has been found to have anticancer effects on several types of cancer cells, but the anticancer effect on breast cancer cells SK-BR-3 has not yet been revealed. Therefore, in this study, DPL caused proliferation inhibition in breast cancer cells SK-BR-3 and the anticancer effect by inducing apoptosis was confirmed, through an in vitro experiment. In order to examine the effect of DPL on cell viability, MTT assay was performed to confirm a significant decrease in the concentration of cell viability. DAPI staining was performed to examine the effect of DPL on cellular morphological changes and increase of apoptotic bodies. To supplement this, an increase in the apoptosis rate was also confirmed through flow cytometry after staining with annexin V/PI. Western blot was performed to confirm apoptosis-related proteins. DPL increased the expression of Cleaved-PARP, Bax whereas decreased the expression of Bcl-2. Changes in the expression levels of MAPK pathway proteins p-ERK1/2, p-JNK, and p-p38 were also confirmed, and a significant increase in p-p38 was observed. These results indicated that DPL induced apoptosis, through p-p38 MAPK signal pathway in SK-BR-3 breast cancer cells.
Ursolic acid is recognized for various effects such as anti-cancer, antioxidant, and anti-inflammatory activity. In this study, we confirmed the anti-cancer effect of ursolic acid on human melanoma cancer cells, A375SM and A375P. Survival rate of the melanoma cells was confirmed by MTT assay and the proliferation rate was confirmed by wound healing assay. The rate of apoptotic bodies was confirmed by DAPI staining, and apoptosis rate was confirmed by flow cytometry. The induction of apoptosis protein was examined by western blotting according to the concentration of ursolic acid in melanoma cells. The survival and proliferation rates of melanoma cells were decreased according to the treatment concentrations of ursolic acid. DAPI staining showed that chromosomal condensation of melanoma cells was increased with increasing concentrations of ursolic acid, and increased apoptosis rate of melanoma cells by ursolic acid was confirmed by flow cytometry. We also confirmed by western blotting that cleaved-PARP and Bax were increased and Bcl-2 was decreased at $12{\mu}M$ concentration of uricolic acid in melanoma cells. This study was carried out at low concentrations of ursolic acid, 0 to $20{\mu}M$, and analyzed 24 h after treatment. As a result of this study, it is thought that ursolic acid has the anti-cancer effect through the regulation of apoptosis-related proteins in melanoma cells A375SM and A375P.
Journal of the Korean Society of Food Science and Nutrition
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v.45
no.5
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pp.671-679
/
2016
Extracts from spinach, cabbage, and onion are known to possess various instructive characteristics, including antioxidant and anti-inflammation activities. Spinach, cabbage, and onion are consumed worldwide and represent important sources of dietary phytochemicals with proven antioxidant properties, such as flavonoids and phenolic acids. Food-derived flavonoids and phenolic compounds are expected to be promising drugs for cancer. In the present study, we investigated the effects of methanol extracts of spinach, cabbage, and onion on cell proliferation and apoptosis in human gastric and breast cancer cells. Proliferation rates of AGS, MDA-MB-231, and SK-BR-3 cells were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The methanol extracts of spinach, cabbage, and onion inhibited proliferation of cancer cells in a dose-dependent manner. 4',6-Diamidino-2-phenylindole (DAPI) staining revealed that chromatin condensation significantly increased compared with the control. In the results of MTT assay and DAPI staining, onion extract was the most effective in inhibiting cancer cell proliferation and apoptosis. To assess changes in protein expression level by onion extract, we identified Bax (pro-apoptotic), Bcl-2 (anti-apoptotic), and poly(ADP-ribose) polymerase (PARP) protein by western blot analysis. The expression of Bax and cleaved-PARP increased, whereas expression of Bcl-2 was decreased compared with the control. These results suggest that spinach, cabbage, and onion extracts suppressed growth of human gastric cancer AGS, human breast cancer MDA-MB-231, and SK-BR-3 cells through induction of apoptosis. Among the extracts, onion extract had stronger anti-cancer and apoptosis induction effects than spinach and cabbage extracts. Further, onion extract more effectively induced apoptosis of human gastric cancer cells than human breast cancer cells. Therefore, further studies are needed to determine the anti-cancer effects of onion extracts in vivo. Onion extract can be developed as a chemopreventive or therapeutic agent for gastric cancer.
Kim, Eun-Ji;Park, So-Young;Hong, Ji-Eun;Shin, Min-Jeong;Lim, Soon-Sung;Shin, Hyun-Kyung;YoonPark, Jung-Han
Journal of the Korean Society of Food Science and Nutrition
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v.36
no.4
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pp.431-438
/
2007
In the present study, twenty eight marine algae species were evaluated for their antiproliferative effect on HT-29 human colon cancer cells. Among these, the methanolic extract of Symphyocladia latiuscula (SL Ex) showed the highest inhibitory activity on HT-29 cell growth. In this study, we examined the mechanism by which SL Ex inhibited the HT-29 cell growth. Cells were cultured with various concentrations of $(0{\sim}20{\mu}g/mL)$ SL Ex. The SL Ex substantially decreased the viable cell numbers and induced apoptosis of HT-29 cells in a dose-dependent manner Western blot analyses of total cell lysates revealed that SL Ex increased the levels of cleaved caspase-8, -9, -7, and -3, and poly (ADP-ribose) polymerase in HT-29 cells. In addition, SL Ex increased truncated Bid levels but moderately decreased Bax levels at only $20{\mu}g/mL$. Furthermore, SL Ex did not affect Bcl-2 protein levels but increased the levels of Fas in HT-29 cells. The present results indicate that SL Ex inhibits cell growth via inducing apoptosis in human colon cancer cells. The mechanism of apoptosis induction by SL Ex involves caspase-8 activation leading to changes in mitochondrial events and subsequent activation of the caspase-7/caspase-3 cascade. Our finding may lead to the development of new therapeutic strategies for the treatment of colon cancer.
Lee, Ji Min;Kim, Hong Jae;Choi, Yung Hyun;Chi, Gyoo Yong;Park, Shin Hyung
Journal of Physiology & Pathology in Korean Medicine
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v.31
no.2
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pp.111-117
/
2017
The anti-cancer effects of Astragalus membranaceus (AM) and Adenophora triphylla var. japonica (AT) have been described. Each of their effects mainly focused on the immunopotentiating and apoptosis inducing-ability in several cancer cell lines. Although the combination of AM and AT is occasionally used in Chinese medicine to treat lung cancers, their synergistic effect has not been proved yet. This study was designed to verify whether AM combined with AT exhibits a synergistic anti-cancer effect in H1299 human lung carcinoma cells. The ethanol extracts of AM (EAM) and AT (EAT) showed only slight cytotoxicity in H1299 cells when treated alone. However, the combination of EAM and EAT markedly suppressed the cell growth measured by MTT assay and trypan blue counting assay. In addition, co-treatment of EAM with EAT significantly reduced the colony-forming ability compared with single treatment of EAM or EAT in H1299 cells. We demonstrated that the synergistic effect of AM and AT was related with apoptosis induction proved by an accumulation of chromatin condensation, annexin V-positive cells, sub-G1 phase population, and cleaved-PARP expression, which were not observed by single treatment of EAM or EAT. In conclusion, the combination of EAM and EAT exhibited superior anti-cancer activity in H1299 cells than single treatment of EAM or EAT. We suggest that EAM combined with EAT might be a novel therapeutic option for lung cancer patients, and provide a reference for the development of more effective combination of Chinese herbs to treat lung cancer.
Kim, Eun-Ji;Lee, Yong-Jin;Shim, Hyun-Kyung;YoonPark, Jung-Han
Journal of the Korean Society of Food Science and Nutrition
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v.35
no.5
/
pp.516-523
/
2006
The mushroom Inonotus obliquue (IO) has been traditionally used for the treatment of gastrointestinal cancer in Russia, Poland, and most of Baltic countries. To explore the possibility that IO has chemoprevention effects, we examined whether or not the aqueous extract of IO inhibits HT-29 cell growth and investigated tile mechanism for this effect. Cells were incubated in the presence of increasing concentrations of the aqueous extract of IO. The extract substantially inhibited the viable HT-29 cell number in a dose-dependent manner and inhibited 5-bromo-2'-deoxyuridine incorporation into DNA of HT-29 cells. Annexin-V staining followed by flow cytometry revealed that the extract induced apoptosis of HT-29 cells in a dose-dependent manner. Western blot analysis of total cell lysates revealed that the extract induced cleavage of caspase-8, -9 and -3 and poly (ADP-ribose) polymerase, but did not affect the protein levels of Bax and Bcl-2. In addition, the extract dose-dependently increased the activity of caspase-8, -9 and -3. We have demonstrated that the aqueous extract of IO inhibits cell proliferation and induces apoptosis in HT-29 cells, which may be mediated by its ability to activate the caspase pathway.
Po, Wah Wah;Thein, Wynn;Khin, Phyu Phyu;Khing, Tin Myo;Han, Khin Wah Wah;Park, Chan Hee;Sohn, Uy Dong
Biomolecules & Therapeutics
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v.28
no.2
/
pp.202-210
/
2020
Fluoxetine is used widely as an antidepressant for the treatment of cancer-related depression, but has been reported to also have anti-cancer activity. In this study, we investigated the cytotoxicity of fluoxetine to human gastric adenocarcinoma cells; as shown by the MTT assay, fluoxetine induced cell death. Subsequently, cells were treated with 10 or 20 µM fluoxetine for 24 h and analyzed. Apoptosis was confirmed by the increased number of early apoptotic cells, shown by Annexin V- propidium iodide staining. Nuclear condensation was visualized by DAPI staining. A significant increase in the expression of cleaved PARP was observed by western blotting. The pan-caspase inhibitor Z-VAD-FMK was used to detect the extent of caspase-dependent cell death. The induction of autophagy was determined by the formation of acidic vesicular organelles (AVOs), which was visualized by acridine orange staining, and the increased expression of autophagy markers, such as LC3B, Beclin 1, and p62/SQSTM 1, observed by western blotting. The expression of upstream proteins, such as p-Akt and p-mTOR, were decreased. Autophagic degradation was evaluated by using bafilomycin, an inhibitor of late-stage autophagy. Bafilomycin did not significantly enhance LC3B expression induced by fluoxetine, which suggested autophagic degradation was impaired. In addition, the co-administration of the autophagy inhibitor 3-methyladenine and fluoxetine significantly increased fluoxetine-induced apoptosis, with decreased p-Akt and markedly increased death receptor 4 and 5 expression. Our results suggested that fluoxetine simultaneously induced both protective autophagy and apoptosis and that the inhibition of autophagy enhanced fluoxetine-induced apoptosis through increased death receptor expression.
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