The aim of this study was to investigate the action of benzalkonium chloride (BAC) used as a preservative in most ophthalmic topical solutions, on human corneal epithelial (HCE) cells in vitro. HCE cell line was exposed to BAC solutions at various concentrations (0.01%~0.0001%) for 15 minutes followed by 24 hours of cell recovery. Cell viability was assessed using MTT assay and chromatin condensation with a Hoechst 33342 test. The expression of membrane protein Fas and Fas ligand was examined by western blot and immunocytochemistry, and DNA fragmentation was studied by agarose gel electrophoresis. A significant decrease of membrane integrity with chromatin condensation was observed with BAC tested at concentrations of 0.005% and higher. BAC was cytotoxic preservatives in this study. An apoptotic mechanism appeared to be present at low concentrations of BAC, whereas a necrotic process appeared at higher concentrations. A functional Fas-mediated apoptotic pathway is present in cultured HCE cells and can be activated by upregulation of Fas expression with BAC.
Oxygen is necessary to sustain life, yet cellular oxygen metabolism creates destructive elements called free radicals. Free radicals are chemically unbalanced and carrying free electrons that can damage molecules, potentially damaging the cell itself. For this reason, many antioxidant products, including supplements and functional foods, are being developed. In particular, natural products are rich sources of pharmacologically active compounds. The purpose of this study was to investigate the antioxidant effects of target biomaterials in Korean traditional spices such as diallyl sulfide (DAS), capsaicin (CAP), and gingerol (GGR), and to investigate the response of the antioxidant defense system to oxidative stress by hydrogen peroxide ($H_2O_2$) compared to sulforaphane (SFN) in HepG2 cells. After the analysis of the cell viability using Cell Counting kit-8 (CCK-8) assay, we determined that the optimum levels were $200{\mu}M$ DAS, $25{\mu}M$ CAP, $50{\mu}M$ GGR, and $12.5{\mu}M$ SFN. Antioxidant enzymes were measured and protein expression was detected by Western blotting. All treatments showed a significant decrease in antioxidant enzyme activity such as superoxide dismutase, catalse, and glutathione peroxidase in HepG2 cells. Additionally, DAS, CAP, GGR and SFN increased the antioxidant system-related transcription factor Nrf2 which was found to be regulated by the activation of MAPK-JNK in this study. In conclusion, these results indicate the protective effects of DAS CAP, GGR, and SFN against $H_2O_2$-induced oxidative stress.
The current study was undertaken to determine the effects of the ethanol extracts of loquat (Eriobotrya japonica Lindl.) seeds, flesh or leaves on the differentiation of 3T3-L1 cells and male pig preadipocytes. The cell number was measured with the MTT assay after trypsin digestion. The cell differentiation was determined by measuring the glycerol-3-phosphate dehydrogenase (GPDH) activity and triglyceride(TG) content. No cytotoxicity was observed from the loquat flesh and leaf ethanol extracts at concentrations of 5, 10, 25, 50, 100 or $200{\mu}g/mL$ in 3T3-L1 cells and pig preadipocytes. However, the cell viability of neither cell line were affected by up $50{\mu}g/mL$ of loquat seed ethanol extract. Treatment with the loquat seed and leaf ethanol extracts significantly suppressed the terminal differentiation of both cell lines in a dose-dependent manner, as confirmed by the decrease in the glycerol-3-phosphate dehydrogenase(GPDH) activity and TG content. Treatment with the loquat seed and leaf ethanol extracts inhibited the GPDH activity and reduced the TG content of both cell types more effectively than that with the loquat flesh ethanol extract. The most potent anti-adipogenic effect was obtained in the case of the ethanol extract of loquat seeds.
Kim, Sung Hak;Choi, Ji Young;Sihn, Choon-Ryoul;Suh, Eun Jung;Kim, Sun Young;Choi, Kang Duk;Jeon, Iksoo;Han, Jae Yong;Kim, Tae-Yoon;Kim, Sang Hoon
Molecules and Cells
/
v.19
no.2
/
pp.185-190
/
2005
The chicken oviduct is a dynamic organ that produces secretory proteins such as ovalbumin and its cells undergo cell proliferation and differentiation. There has been no study of the cellular mechanism involved in cell death in the chicken oviduct. Therefore, this study has focused on the study of apoptosis in primary oviduct cells. Because ceramide is known to activate apoptosis in tumor cells and is produced in the oviduct, we used an exogenous ceramide analog to induce cell death. The viability of ceramide-treated chicken oviduct cells decreased in a dose-dependent manner and apoptotic cells were detected by staining with annexin V. The expression of apoptosis-related genes was assessed by RT-PCR and bcl-2 mRNA was found to decrease after exposure to ceramide while Bcl-x mRNA increased 12 h post-treatment. In addition, caspase-3 was expressed strongly in the early stages of apoptosis, while caspase-1 and -9 transcripts increased at later times. We conclude that ceramide induces apoptosis in oviduct-derived primary cells via a caspase- and bcl-2-dependent pathway.
$Amyloid-{\beta}$ protein ($A{\beta}$) is known to increase free radical production in neuronal cells, leading to cell death by oxidative stress. The purpose of this study was to evaluate the protective effects of $PineXol^{(R)}$ on $A{\beta}_{25-35}$ induced neuronal cell death. Rat pheochromocytoma (PC-12) cells were pre-treated with $100{\mu}g/mL$ of $PineXol^{(R)}$ for 2 h. The cells were exposed to single dose of $30{\mu}M$$A{\beta}_{25-35}$ for 24 h. Cell death was assessed by a cell count kit-8 (CCK-8) assay, lactate and dehydrogenase (LDH) release assay. An Apoptotic process was analyzed by a protein expression of the Bcl-2 family using western blotting. Cell viability increased in PC-12 cells treated with both $A{\beta}_{25-35}$ and $PineXol^{(R)}$, compared to the control group. $PineXol^{(R)}$ induced a decrease of the Bcl-2 protein expression (p<0.05), while Bax and Sod1 increased (p<0.05), indicating attenuation of $A{\beta}_{25-35}$ induced apoptosis. These results suggest that $PineXol^{(R)}$ may be a good candidate for the prevention of Alzheimer's disease(AD).
Kim, Hong Jae;Jeong, Jin-Woo;Park, Cheol;Choi, Yung Hyun;Hong, Su Hyun
Journal of Pharmacopuncture
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v.22
no.4
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pp.269-278
/
2019
Objectives: Naesohwangryeon-tang (NHT) is a type of traditional herbal formula, however, little is known about its antitumor activity. In this study, the antitumor properties of NHT was evaluated in human lung adenocarcinoma cells. Methods: To check the inhibitory effect of NHT, MTT assay was performed. Cell cycle analysis and detection of ROS production were conducted by flow cytometry. To evaluate the signaling pathway, Western blotting was conducted. Results: Our results showed that the decrease of cell proliferation by NHT stimulation occurred more significantly in A549 cells than in NCI-H460 cells. In addition, NHT-induced apoptosis was associated with the activation of caspases and production of reactive oxygen species (ROS). NHT-induced apoptosis was attenuated after pretreatments with z-VAD-fmk or N-acetylcysteine, suggesting that NHT-induced apoptosis was caspaseand ROS-dependent. Interestingly, NHT treatment led to the development of autophagic vesicular organelles and upregulation of several autophagy-related genes. The pretreatment of bafilomycin A1 decreased apoptosis slightly but increased cell viability in the presence of NHT. Conclusion: These findings indicated that NHT induces both apoptosis and cell-protective autophagy in human lung cancer cells. This data suggests that NHT might be a novel herbal drug for lung cancer.
Objective: This study aimed to investigate the trend in the research on breast cancer using traditional Korean medicine (TKM) and establish the direction for further study. Methods: Breast cancer studies using Korean medicine were searched using the Oriental Medicine Advanced Searching Integrated System (OASIS). The search term was 'breast' and there was no restriction in year. The searched studies were analyzed according to the type of research. Results: 1. 83 studies were searched. The types and numbers of study were as follows: 42 were in vitro studies, 5 were in vivo studies, 12 were studies for review, and 27 were clinical research including case reports. 2. Various cell lines such as MCF-7, MDA-MB-231, SKBR3, and MCF-10A were used for in vitro studies. The studies reported a decrease in cell viability, induction of apoptosis, and change of expression in cancer-related genes. In vivo studies also reported induction of apoptosis, and anti-proliferative activity of herbal medicine against the cancer cells. 3. Among the clinical research, 8 were cross-sectional studies, 3 were controlled-trial, and 15 were case reports. The baseline characteristics of breast cancer patients were analyzed in the cross-sectional studies. Interventions such as pharmacopuncture, herbal medicine, massage, Qi gong, acupuncture, electroacupuncture and moxibustion were used in clinical research. 4. Research on the review of breast cancer covered various subjects as follows: herbal medicine, acupuncture, pattern identification of breast cancer in traditional Korean medicine, analysis of previous experimental studies, and clinical trials. Conclusion: We have found the applicability of TKM for treatment of breast cancer through this review. It is necessary to conduct further studies, such as well-designed clinical trials based on the results from experimental research.
Journal of Physiology & Pathology in Korean Medicine
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v.31
no.1
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pp.52-58
/
2017
This study was aimed to examine relaxing effects of Acanthopanacis cortex(AC) through nitric oxide(NO) production and phosphodiesterase type 5(PDE-5) inhibition in corpus cavernosum. In order to define the relaxation effects of AC extract, rabbit corpus cavernous tissues were prepared in $2{\times}2{\times}8mm$ sized strip. AC extract ($0.01-3.0mg/m{\ell}$) were treated in contracted strips induced by phenylephrine(PE) and $N{\omega}$-nitro-L-arginine (L-NNA) was treated before AC extract-treated. And calcium chloride($Ca^{2+}$) 1 mM was infused into precontracted strips after pretreatment of AC extract in $Ca^{2+}-free$ krebs-ringer solution. When AC extract was applied to human umbilical vein endothelial cell(HUVEC), cell viability was measured by MTT assay, and NO concentration was measured by Griess reagent system. Ratio of smooth muscles to collagen fibers and eNOS, PDE-5 positive reaction were measured by histochemical and immunohistochemical process on mice corpus cavernosum. AC extract significantly affected relaxion of the cavernous strips, and the pretreatment of L-NNA inhibited AC extract-induced relaxation. Contraction induced by the addition of $Ca^{2+}$ was inhibited by treatment with the AC extract in $Ca^{2+}-free$ solution. In AC group, NO concentration, ratio of smooth muscle to collagen fibers, and eNOS positive reaction were increased, PDE-5 positive reaction was decreased compared to PE group. As a result of the above experiment, it was thought that AC extract inhibits the inflow of extracellular $Ca^{2+}$ by activating cGMP through the increase of eNOS / NO and the decrease of PDE-5 which inhibits cGMP activity, in the corpus cavernosum.
Kim, Mi Jin;Kadayat, Taraman;Um, Yeon Ji;Jeong, Tae Cheon;Lee, Eung-Seok;Park, Pil-Hoon
Biomolecules & Therapeutics
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v.23
no.2
/
pp.119-127
/
2015
Chalcones (1,3-diaryl-2-propen-1-ones), a subfamily of flavonoid, are widely known to possess potent anti-inflammatory and anti-oxidant properties. In this study, we investigated the effect of 3-(4-Hydroxyphenyl)-1-(thio3-(4-Hydroxyphenyl phen-2-yl)prop-2-en-1-one (TI-I-175), a synthetic chalcone derivative, on endotoxin-induced expression of monocyte chemoattractant protein-1 (MCP-1), one of the key chemokines that regulates migration and infiltration of immune cells, and its potential mechanisms. TI-I-175 potently inhibited MCP-1 mRNA expression stimulated by lipopolysaccharide (LPS) in RAW 264.7 macrophages without significant effect on cell viability. Treatment of cells with TI-I-175 markedly prevented LPS-induced transcriptional activation of activator protein-1 (AP-1) as measured by luciferase reporter assay, while nuclear factor-${\kappa}B$ (NF-${\kappa}B$) activity was not inhibited by TI-I-175, implying that TI-I-175 suppressed MCP-1 expression probably via regulation of AP-1. In addition, TI-I-175 treatment significantly inhibited LPS-induced Akt phosphorylation and led to a significant decrease in reactive oxygen species (ROS) production by LPS, which act as up-stream signaling events required for AP-1 activation in RAW 264.7 macrophages. Taken together, these results indicate that TI-I-175 suppresses MCP-1 gene expression in LPS-stimulated RAW 264.7 macrophages via suppression of ROS production and Akt activation.
Kim, Jong-Man;Lee, Jae-Yeon;Kim, Duck-Hwan;Jeong, Seong-Mok;Park, Chang-Sik;Kim, Myung-Cheol
Journal of Veterinary Clinics
/
v.25
no.3
/
pp.165-169
/
2008
Renal ischemia-reperfusion (I/R) injury is great clinical important because viability of the organ depends on the tolerance to ischemia-reperfusion injury, an inevitable processing during surgery. The purpose of this study was to investigate the effects of premedicated ascorbic acid alone in I/R injury model induced by cross-clamping of renal vessels. In the rabbit models, 2-4 kg New Zealand white rabbits were subjected to 30 minutes of warm unilateral renal ischemia followed by removal of contralateral kidney and then divided into five groups, control (2) arid treatment groups (3). In control group 1, the rabbits only received right nephrectomy. In control group 2, the rabbits received I/R on left kidney after the right nephrectomy. In treatment group 1, the rabbits received ascorbic acid 50 mg/kg IV before the operation. In treatment group 2, the rabbits received ascorbic acid 100 mg/kg IV before the operation. In treatment group 3, the rabbits received ascorbic acid 200 mg/kg IV before the operation. Blood samples were collected from these rabbits for measurement of kidney function tests at the 0, 1 st, 3rd and 7th day and antioxidant enzyme( SOD, GSHPx, CAT) at 24 hours. Kidney function tests (serum creatinine and BUN) showed a significant difference between group 2 and group 4, 5. Activity of antioxidant enzymes in plasma were significant decrease in group 4, 5 compare to group 2. The result of this study suggested that the exogenous ascorbic acid had a role of attenuation of renal I/R injury in rabbit model.
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