• YAKHAK HOEJI
• /
• v.50 no.4
• /
• pp.272-277
• /
• 2006

This study was undertaken to determine whether the 9 herbal complex induces apoptosis in human breast cancer MCF-7 cells and adriamycin-resistant MCF7/adR cells. Ethanol extracts of Bojungbangamtang (BBTE) and acidic polysaccharide of Red Ginseng (GIN) induced cell death in both MCF-7 and MCF7/adR cells. Ethanol extracts of Bojungbangamtang and acidic polysaccharide of Red Ginseng also induced $G_2/M$ cell cycle arrest and increased TUNEL positive cells in MCF7/adR cells. In addition, flow cytometric analysis revealed the decreased expression of P-glycoprotein (P-gp) in ethanol extracts of Bojungbangamtang and acidic polysaccharide of Red Ginseng treated MCF7/adR cells. Similarly, decreased protein levels of P-glycoprotein and multidrug resistance associated proteins-1 were also determined by immunocytometry in ethanol extracts of Bojungbangamtang treated MCF7/adR cells. Taken together these data indicate that ethanol extracts of Bojungbangamtang and acidic polysaccharide of Red Ginseng inhibit the function of ABC transporters such as multi drug resistance associated proteins (MRPs) and P-glycoprotein as well as induce apoptosis in MCF7/adR cells. Thus, these data suggest that ethanol extracts of Bojungbangamtang and polysaccharide of Red Ginseng can be candidates for the treatment of multidrug-resistant MCF7/adR cells.

• Journal of the Korea Academia-Industrial cooperation Society
• /
• v.20 no.2
• /
• pp.286-293
• /
• 2019

Alzheimer's disease (AD) is a neurodegenerative disease caused by accumulation of amyloid beta ($A{\beta}$) in the brain. In the present study, we investigated the neuroprotective effects of four flavonoids such as kaempferol, kaempferol-3-O-glucoside, quercetin, and quercetin-3-${\beta}$-D-glucoside against neuronal apoptosis induced by $A{\beta}$ in SH-SY5Y neuronal cells. Treatment with $A{\beta}$ decreased cell viability compared to the non-treated normal group. However, treatment with the four flavonoids increased cell viability in SH-SY5Y cells treated with $A{\beta}$. In addition, we measured the expression of apoptosis-related proteins such as Bcl-2-associated X protein (Bax) and cleaved caspase-9. Treatment with the four flavonoids down-regulated Bax and cleaved caspase-9 in $A{\beta}$-treated SH-SY5Y neuronal cells. Overall, the results of the present study demonstrated the neuroprotective effect of flavonoids by anti-apoptotic activity in $A{\beta}$-induced SH-SY5Y neuronal cells. These results suggest that these four flavonoids would be useful therapeutic and prevention agents for AD.

• Journal of Life Science
• /
• v.29 no.5
• /
• pp.514-523
• /
• 2019

We investigated the gene expression patterns and the mechanisms of action of the apoptotic response by microarray analysis of human keratinocyte HaCaT cells treated with ginsenoside Rb1, a saponin of Panax ginseng C. A. Meyer. Genes related to apoptosis, the G2/M transition of the mitotic cell cycle, cell division, mitotic nuclear division, and intracellular protein transport were 2-fold up-regulated in HaCaT cells treated with the ginsenoside Rb1, whereas genes related to DNA repair, regeneration fission, and extracellular matrix organization were 2-fold down-regulated. Apoptosis signaling may be mediated by FAS and PLA2G4A, and pathway analysis indicated that STAT3 might be an upstream regulator of these genes. The activity of FAS and PLA2G4A was verified by qPCR, which showed that FAS was increased about 2-fold in HaCaT cells treated with $10{\mu}g/ml$ of ginsenoside Rb1 for 24 hr, PLA2G4A was increased about twice after 6 hours, and gene expression was increased more than 2-fold after 24 hr. Knockdown of STAT3 with siRNA decreased FAS expression and increased PLA2G4A expression but only FAS was passed from the upstream regulator STAT3. These results indicate that STAT3, which is an upstream regulator, induces apoptosis via FAS during treatment with ginsenoside Rb1.

• Yonsei Medical Journal
• /
• v.59 no.9
• /
• pp.1096-1106
• /
• 2018

Purpose: Alzheimer's disease (AD) is the sixth most common cause of death in the United States. MicroRNAs have been identified as vital players in neurodegenerative diseases, including AD. microRNA-128 (miR-128) has been shown to be dysregulated in AD. This study aimed to explore the roles and molecular mechanisms of miR-128 in AD progression. Materials and Methods: Expression patterns of miR-128 and peroxisome proliferator-activated receptor gamma ($PPAR-{\gamma}$) messenger RNA in clinical samples and cells were measured using RT-qPCR assay. $PPAR-{\gamma}$ protein levels were determined by Western blot assay. Cell viability was determined by MTT assay. Cell apoptotic rate was detected by flow cytometry via double-staining of Annexin V-FITC/PI. Caspase 3 and $NF-{\kappa}B$ activity was determined by a Caspase 3 Activity Assay Kit or $NF-{\kappa}B$ p65 Transcription Factor Assay Kit, respectively. Bioinformatics prediction and luciferase reporter assay were used to investigate interactions between miR-128 and $PPAR-{\gamma}$ 3'UTR. Results: MiR-128 expression was upregulated and $PPAR-{\gamma}$ expression was downregulated in plasma from AD patients and $amyloid-{\beta}$ $(A{\beta})-treated$ primary mouse cortical neurons (MCN) and Neuro2a (N2a) cells. Inhibition of miR-128 decreased $A{\beta}-mediated$ cytotoxicity through inactivation of $NF-{\kappa}B$ in MCN and N2a cells. Moreover, $PPAR-{\gamma}$ was a target of miR-128. $PPAR-{\gamma}$ upregulation attenuated $A{\beta}-mediated$ cytotoxicity by inactivating $NF-{\kappa}B$ in MCN and N2a cells. Furthermore, $PPAR-{\gamma}$ downregulation was able to abolish the effect of anti-miR-128 on cytotoxicity and $NF-{\kappa}B$ activity in MCN and N2a cells. Conclusion: MiR-128 inhibitor decreased $A{\beta}-mediated$ cytotoxicity by upregulating $PPAR-{\gamma}$ via inactivation of $NF-{\kappa}B$ in MCN and N2a cells, providing a new potential target in AD treatment.

• Korean Journal of Food Science and Technology
• /
• v.54 no.2
• /
• pp.134-140
• /
• 2022

Kelp belongs to the brown algae family and has been reported to exert anti-cancer effects on some cancer types, however studies have not been reported on the anti-cancer effects of kelp extracts on melanoma. In this study, the anti-cancer effects of kelp extract in B16-F0 cells were investigated, and the underlying molecular mechanisms were assessed. Kelp extract was found to inhibit the proliferation of B16-F0 cells, induce cytotoxicity, inhibit cell colony formation, and induce DNA fragmentation and apoptosis. The molecular mechanism was found to involve kelp extract increasing the expression of cytochrome-c and activated caspase-9 in the intrinsic apoptotic pathway. In addition, kelp extract upregulated the expression of Fas-associated protein with death domain and activated caspase-8 in the extrinsic apoptosis pathway. Activation of caspase-9 and caspase-8 by kelp extract induced activation of caspase-3 and cleaved poly adenosine diphosphate-ribose polymerase, consequently inducing apoptosis. These data suggest that kelp extract represents a potential therapeutic agent for melanoma.

• Journal of Life Science
• /
• v.31 no.4
• /
• pp.389-399
• /
• 2021

This study determined the effects of Centella asiatica leaf on H₂O₂ induced cell cycle arrest, mitochondrial activity, and proinflammatory cytokine production in human dermal fibroblast (HDF) cells. We used an 80% methanol extract of C. asiatica, its ethyl acetate fraction, and asiaticoside, the major constituent of C. asiatica. The C. asiatica extract, its ethyl acetate fraction, and asiaticoside attenuated G1 cell cycle-arrest and the apoptotic effect caused by H₂O₂-induced oxidative stress. The cells treated with C. asiatica extract, its ethyl acetate fraction, and asiaticoside secreted lower levels of TNF-α and IL-6. The antioxidant effect of asiaticoside was higher than that of C. asiatica extract and its ethyl acetate fraction. Treatment with C. asiatica extract, its ethyl acetate fraction, and asiaticoside also increased the mitochondrial membrane potential and restored normal mitochondrial morphology. Following H₂O₂ stress induction, cells treated with C. asiatica extract, its ethyl acetate fraction, and asiaticoside showed increased mitochondrial oxygen consumption rates and decreases in the TLR4-MyD88-TRAF6-p65 pathway activity. These findings suggest that C. asiatica extract, its ethyl acetate fraction, and asiaticoside have antioxidant and anti-inflammatory effects, as well as the ability to control the mitochondrial activities of HDF cells.

• Tuberculosis and Respiratory Diseases
• /
• v.43 no.1
• /
• pp.63-68
• /
• 1996

Background: Interferon-$\gamma$ has various biologic effects, including antiviral effect, antitumor proliferative effect, activation of macrophage and B lymphocyte, and increased expression of major histocompatibility complex. Especially, antitumor proliferative effect of interferon-$\gamma$ has already been proved to be important in vivo as well as in vitro. And, clinical studies of interferon-$\gamma$ have been tried in lung cancer patients. However, the mechanism of antitumor effect of interferon-$\gamma$ has not yet been established despite of many hypotheses. "Necrosis" is a type of cell death which is well known to occur in the circumstances of severe stresses. In contrast, "apoptosis" is another type of cell death which occurs in such biological circumstances as embryonic development, regression of organs, and self-tolerance of lymphocytes. And, apoptosis is an active process of cell death in which cells are dying with fragmentations of their cytoplasms and nuclei. And, in the process of apoptosis the DNAs of cells are cleaved between nucleosomes by unidentified endonuclease and therefore DNAs of apoptotic cells result in a typical electrophoresis pattern known as DNA ladder pattern. Recently it has been suggested that cytotoxic effect of interferon-$\gamma$ occurs via apoptosis. To elucidate the mechanism of antitumor cytotoxic effect of interferon-$\gamma$, we microscopically observed a lung cancer cell line, A549 which was treated with interferon-$\gamma$. We observed A545 treated with interferon-$\gamma$ was dying fragmented. And so, we performed this study to find out that the mechanism of antitumor cytotoxic effect of interferon-$\gamma$ be apoptosis. Method: We treated A549, human lung cancer cell line with various concentration of interferon-$\gamma$ and quantified its cytotoxic effect of various periods, 24 hours, 72 hours and, 120 hours by MTT(dimethylthiazolyl diphenyltetrazolium bromide) bioassay. Also, after we treated A549 with 100 units/mi of interferon-$\gamma$ for 120 hours, we observed the pattern of cell death with inverted microscope and we extracted DNAs from the dead A549 cells and observed the pattern of 1.5% agarose gel electrophoresis with ethidium bromide staining. Result: 1) Cytotoxic effect of interferon-$\gamma$ on A549: For the first 24 hours, threre was little cytotoxic effect and for between 24 hours and 72 hours, there was the beginning of cytotoxic effect and for 120 hours there was increased cytotoxic effect. 2) Pattern of A549 cell death by interferon-$\gamma$: We observed with inverted microscope that A549 cells were dying fragmented. 3) DNA ladder pattern of gel electrophoresis: We observed DNA ladder pattern of gel electrophoresis of extracted DNAs from dead A549 cells. Conclusion: We concluded that the mechanism of interferon-$\gamma$induced cytotoxicity on lung cancer cell line, A549 be via apoptosis.

• Journal of the Society of Cosmetic Scientists of Korea
• /
• v.42 no.1
• /
• pp.75-85
• /
• 2016

Pectin, a naturally occurring polysaccharide, has in recent years attracted considerable attention. Its benefits are increasingly appreciated by scientists and consumers due to its safety and usefulness. The chemistry and gel-forming characteristics of pectin have enabled to be used in pharmaceutical industry, health promotion and treatment. Yet, it has been rarely used in cosmetics because of its incompatibility with many cosmetic ingredients, including alcohols, and unstable viscosity of pectin gels under various pH and salt conditions. However, low-molecular-weight pectin oligomers have excellent biological activities, and depolymerization of pectin to produce cosmetic ingredients would be very useful. In this study, we attempted the development of cosmetic ingredients using pectin with an excellent effect on human skin. We developed a bio-conversion process that uses enzymatic hydrolysis to produce pectin hydrolysates containing mainly low-molecular-weight pectin oligomers. Gel permeation chromatography was used to determined the ratio of hydrolysis. The molecular weight of the pectin hydrolysates obtained varied between 200 and 2,700 Da. The two newly developed low-molecular-weight pectin hydrolysates, LMPH A and B, had higher anti-oxidative activities than pectin or D-galacturonic. Exposure to UVB radiation induces apoptotic cell death in epidermal cells. Annexin V binding and propidium iodide uptake were measured by flow cytometry to evaluate UVB-induced cell death in HaCaT cells. Both LMPH A and B reduced UVB-induced cell death and increased cell proliferation by 22% and 30% at 0.5% concentration respectively, while pectin had no significant activity. In conclusion, this study suggests that the newly developed low-molecular-weight pectin hydrolysates can be used as safe and biologically active cosmetic ingredients.

• The Korean Journal of Nuclear Medicine
• /
• v.37 no.3
• /
• pp.190-198
• /
• 2003

Purpose: To evaluate radiation sensitivity of dendritic cells in comparison with lymphocytes. Materials and methods: T lymphocytes captured from peripheral blood were irradiated by 0 Gy, 10 Gy, 30 Gy. Apoptosis was measured by flowcytometry for staining of Annexin V 4 hours after irradiation. Immature and mature dendritic cells processed from blood hematopoietic stem cell were irradiated by 0 Gy, 10 Gy, 30 Gy, 100 Gy respectively and apoptosis was measured by flowcytometry with time difference as 4h, 24h and 48h after irradiation. Morphometric analysis by percent nucleus was measured in three cell groups, also. Results: Lymphocytes showed radiation sensitivity by increasing apoptotic fraction according to radiation dose. However, both mature and immature dendritic cells showed consistent fraction of apoptosis in spite of increasing radiation dose. Percent nucleus ratio is significantly higher in lymphocytes than that of mature or immature dendritic cells. Stimulation of T-cell by dendritic cells was not changed after irradiation. Conclusion: Dendritic cells showed radioresistance which was associated with small size of nucleus in comparison with lymphocytes and this result would be used as a basal data of radio-labelling for the cellular trafficking studios in nuclear medicine fields.

• Journal of Nutrition and Health
• /
• v.50 no.5
• /
• pp.415-425
• /
• 2017

Purpose: Many studies have suggested that neuronal cells protect against oxidative stress-induced apoptotic cell death by polyphenolic compounds. We investigated the neuroprotective effects and the mechanism of action of Momordica charantia ethanol extract (MCE) against $H_2O_2-induced$ cell death of human neuroblastoma SK-N-MC cells. Methods: The antioxidant activity of MCE was measured by the quantity of total phenolic acid compounds (TPC), quantity of total flavonoid compounds (TFC), and 2,2-Diphenyl-1-pycrylhydrazyl (DPPH) radical scavenging activity. Cytotoxicity and cell viability were determined by CCK-8 assay. The formation of reactive oxygen species (ROS) was measured using 2,7-dichlorofluorescein diacetate (DCF-DA) assay. Antioxidant enzyme (SOD-1,2 and GPx-1) expression was determined by real-time PCR. Mitogen-activated protein kinases (MAPK) pathway and apoptosis signal expression was measured by Western blotting. Results: The TPC and TFC quantities of MCE were 28.51 mg gallic acid equivalents/extract g and 3.95 mg catechin equivalents/extract g, respectively. The $IC_{50}$ value for DPPH radical scavenging activity was $506.95{\mu}g/ml$ for MCE. Pre-treatment with MCE showed protective effects against $H_2O_2-induced$ cell death and inhibited ROS generation by oxidative stress. SOD-1,2 and GPx-1 mRNA expression was recovered by pre-treatment with MCE compared with the presence of $H_2O_2$. Pre-treatment with MCE inhibited phosphorylation of p38 and the JNK pathway and down-regulated cleaved caspase-3 and cleaved PARP by $H_2O_2$. Conclusion: The neuroprotective effects of MCE in terms of recovery of antioxidant enzyme gene expression, down-regulation of MAPK pathways, and inhibition apoptosis is associated with reduced oxidative stress in SK-N-MC cells.