• Biomolecules & Therapeutics
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• v.29 no.2
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• pp.227-233
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• 2021

Benzo[a]pyrene (B[a]P) is a polycyclic aromatic hydrocarbon and ubiquitous environmental toxin with known harmful effects to human health. Abnormal phenotypes of keratinocytes are closely associated with their exposure to B[a]P. Resorcinol is a component of argan oil with reported anticancer activities, but its mechanism of action and potential effect on B[a]P damage to the skin is unknown. In this study, we investigated the effects of resorcinol on B[a]P-induced abnormal keratinocyte biology and its mechanisms of action in human epidermal keratinocyte cell line HaCaT. Resorcinol suppressed aryl hydrocarbon receptor (AhR) activity as evidenced by the inhibition of B[a]P-induced xenobiotic response element (XRE)-reporter activation and cytochrome P450 1A1 (CYP1A1) expression. In addition, resorcinol attenuated B[a]P-induced nuclear translocation of AhR, and production of ROS and pro-inflammatory cytokines. We also found that resorcinol increased nuclear factor (erythroid-derived 2)-like 2 (Nrf2) activity. Antioxidant response element (ARE)-reporter activity and expression of ARE-dependent genes NAD(P)H dehydrogenase [quinone] 1 (NQO1), heme oxygenase-1 (HO-1) were increased by resorcinol. Consistently, resorcinol treatment induced nuclear localization of Nrf2 as seen by Western analysis. Knockdown of Nrf2 attenuated the resorcinol effects on ARE signaling, but knockdown of AhR did not affect resorcinol activation of Nrf2. This suggests that activation of antioxidant activity by resorcinol is not mediated by AhR. These results indicate that resorcinol is protective against effects of B[a]P exposure. The mechanism of action of resorcinol is inhibition of AhR and activation of Nrf2-mediated antioxidant signaling. Our findings suggest that resorcinol may have potential as a protective agent against B[a]P-containing pollutants.

• Journal of Ginseng Research
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• v.45 no.3
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• pp.390-400
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• 2021

Background: We recently showed that gintonin, an active ginseng ingredient, exhibits antibrain neurodegenerative disease effects including multiple target mechanisms such as antioxidative stress and antiinflammation via the lysophosphatidic acid (LPA) receptors. Amyotrophic lateral sclerosis (ALS) is a spinal disease characterized by neurodegenerative changes in motor neurons with subsequent skeletal muscle paralysis and death. However, pathophysiological mechanisms of ALS are still elusive, and therapeutic drugs have not yet been developed. We investigate the putative alleviating effects of gintonin in ALS. Methods: The G93A-SOD1 transgenic mouse ALS model was used. Gintonin (50 or 100 mg/kg/day, p.o.) administration started from week seven. We performed histological analyses, immunoblot assays, and behavioral tests. Results: Gintonin extended mouse survival and relieved motor dysfunctions. Histological analyses of spinal cords revealed that gintonin increased the survival of motor neurons, expression of brain-derived neurotrophic factors, choline acetyltransferase, NeuN, and Nissl bodies compared with the vehicle control. Gintonin attenuated elevated spinal NAD(P) quinone oxidoreductase 1 expression and decreased oxidative stress-related ferritin, ionized calcium-binding adapter molecule 1-immunoreactive microglia, S100β-immunoreactive astrocyte, and Olig2-immunoreactive oligodendrocytes compared with the control vehicle. Interestingly, we found that the spinal LPA1 receptor level was decreased, whereas gintonin treatment restored decreased LPA1 receptor expression levels in the G93A-SOD1 transgenic mouse, thereby attenuating neurological symptoms and histological deficits. Conclusion: Gintonin-mediated symptomatic improvements of ALS might be associated with the attenuations of neuronal loss and oxidative stress via the spinal LPA1 receptor regulations. The present results suggest that the spinal LPA1 receptor is engaged in ALS, and gintonin may be useful for relieving ALS symptoms.

• The Korean Journal of Physiology and Pharmacology
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• v.26 no.4
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• pp.263-275
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• 2022

There is a paucity of detailed data related to the effect of senescence on the mitochondrial antioxidant capacity and redox state of senescent human cells. Activities of TCA cycle enzymes, respiratory chain complexes, hydrogen peroxide (H₂O₂), superoxide anions (SA), lipid peroxides (LPO), protein carbonyl content (PCC), thioredoxin reductase 2 (TrxR2), superoxide dismutase 2 (SOD2), glutathione peroxidase 1 (GPx1), glutathione reductase (GR), reduced glutathione (GSH), and oxidized glutathione (GSSG), along with levels of nicotinamide cofactors and ATP content were measured in young and senescent human foreskin fibroblasts. Primary and senescent cultures were biochemically identified by monitoring the augmented cellular activities of key glycolytic enzymes including phosphofructokinase, lactate dehydrogenase, and glycogen phosphorylase, and accumulation of H₂O₂, SA, LPO, PCC, and GSSG. Citrate synthase, aconitase, α-ketoglutarate dehydrogenase, succinate dehydrogenase, malate dehydrogenase, isocitrate dehydrogenase, and complex I-III, II-III, and IV activities were significantly diminished in P25 and P35 cells compared to P5 cells. This was accompanied by significant accumulation of mitochondrial H₂O₂, SA, LPO, and PCC, along with increased transcriptional and enzymatic activities of TrxR2, SOD2, GPx1, and GR. Notably, the GSH/GSSG ratio was significantly reduced whereas NAD⁺/NADH and NADP⁺/NADPH ratios were significantly elevated. Metabolic exhaustion was also evident in senescent cells underscored by the severely diminished ATP/ADP ratio. Profound oxidative stress may contribute, at least in part, to senescence pointing at a potential protective role of antioxidants in aging-associated disease.

• Journal of Ginseng Research
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• v.46 no.6
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• pp.759-770
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• 2022

Background: Aerobic cellular respiration provides chemical energy, adenosine triphosphate (ATP), to maintain multiple cellular functions. Sirtuin 1 (SIRT1) can deacetylate peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC-1α) to promote mitochondrial biosynthesis. Targeting energy metabolism is a potential strategy for the prevention and treatment of various diseases, such as cardiac and neurological disorders. Ginsenosides, one of the major bioactive constituents of Panax ginseng, have been extensively used due to their diverse beneficial effects on healthy subjects and patients with different diseases. However, the underlying molecular mechanisms of total ginsenosides (GS) on energy metabolism remain unclear. Methods: In this study, oxygen consumption rate, ATP production, mitochondrial biosynthesis, glucose metabolism, and SIRT1-PGC-1α pathways in untreated and GS-treated different cells, fly, and mouse models were investigated. Results: GS pretreatment enhanced mitochondrial respiration capacity and ATP production in aerobic respiration-dominated cardiomyocytes and neurons, and promoted tricarboxylic acid metabolism in cardiomyocytes. Moreover, GS clearly enhanced NAD⁺-dependent SIRT1 activation to increase mitochondrial biosynthesis in cardiomyocytes and neurons, which was completely abrogated by nicotinamide. Importantly, ginsenoside monomers, such as Rg1, Re, Rf, Rb1, Rc, Rh1, Rb2, and Rb3, were found to activate SIRT1 and promote energy metabolism. Conclusion: This study may provide new insights into the extensive application of ginseng for cardiac and neurological protection in healthy subjects and patients.

• Biomolecules & Therapeutics
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• v.31 no.1
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• pp.73-81
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• 2023

Sirtuins (SIRTs) belong to the nicotinamide adenine dinucleotide (NAD+)-dependent class III histone deacetylase family. They are key regulators of cellular and physiological processes, such as cell survival, senescence, differentiation, DNA damage and stress response, cellular metabolism, and aging. SIRTs also influence carcinogenesis, making them potential targets for anticancer therapeutic strategies. In this study, we investigated the anticancer properties and underlying molecular mechanisms of a novel SIRT1 inhibitor, MHY2251, in human colorectal cancer (CRC) cells. MHY2251 reduced the viability of various human CRC cell lines, especially those with wild-type TP53. MHY2251 inhibited SIRT1 activity and SIRT1/2 protein expression, while promoting p53 acetylation, which is a target of SIRT1 in HCT116 cells. MHY2251 treatment triggered apoptosis in HCT116 cells. It increased the percentage of late apoptotic cells and the sub-G1 fraction (as detected by flow cytometric analysis) and induced DNA fragmentation. In addition, MHY2251 upregulated the expression of FasL and Fas, altered the ratio of Bax/Bcl-2, downregulated the levels of pro-caspase-8, -9, and -3 proteins, and induced subsequent poly(ADP-ribose) polymerase cleavage. The induction of apoptosis by MHY2251 was related to the activation of the caspase cascade, which was significantly attenuated by pre-treatment with Z-VAD-FMK, a pan-caspase inhibitor. Furthermore, MHY2251 stimulated the phosphorylation of c-Jun N-terminal kinase (JNK), and MHY2251-triggered apoptosis was blocked by pre-treatment with SP600125, a JNK inhibitor. This finding indicated the specific involvement of JNK in MHY2251-induced apoptosis. MHY2251 shows considerable potential as a therapeutic agent for targeting human CRC via the inhibition of SIRT1 and activation of JNK/p53 pathway.

• Journal of Veterinary Science
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• v.24 no.2
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• pp.24.1-24.13
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• 2023

Background: Ergothioneine (EGT) is a natural amino acid derivative in various animal organs and is a bioactive compound recognized as a food and medicine. Objectives: This study examined the effects of EGT supplementation during the in vitro maturation (IVM) period on porcine oocyte maturation and subsequent embryonic development competence after in vitro fertilization (IVF). Methods: Each EGT concentration (0, 10, 50, and 100 μM) was supplemented in the maturation medium during IVM. After IVM, nuclear maturation, intracellular glutathione (GSH), and reactive oxygen species (ROS) levels of oocytes were investigated. In addition, the genes related to cumulus function and antioxidant pathways in oocytes or cumulus cells were investigated. Finally, this study examined whether EGT could affect embryonic development after IVF. Results: After IVM, the EGT supplementation group showed significantly higher intracellular GSH levels and significantly lower intracellular ROS levels than the control group. Moreover, the expression levels of hyaluronan synthase 2 and Connexin 43 were significantly higher in the 10 μM EGT group than in the control group. The expression levels of nuclear factor erythroid 2-related factor 2 (Nrf2) and NAD(P)H quinone dehydrogenase 1 (NQO1) were significantly higher in the oocytes of the 10 μM EGT group than in the control group. In the assessment of subsequent embryonic development after IVF, the 10 μM EGT treatment group improved the cleavage and blastocyst rate significantly than the control group. Conclusions: Supplementation of EGT improved oocyte maturation and embryonic development by reducing oxidative stress in IVM oocytes.

• Journal of Acupuncture Research
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• v.20 no.5
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• pp.118-132
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• 2003

Objective: The aim of this study was to investigate the effects of Joksamni(ST36) combination on NAD PH-diaphorase, neuronal nitric oxide synthase(nNOS), neuropeptide Y(NPY) and vasoactive intestinal peptide (VIP) in the cerebral cortex of spontaneously hypertensive rat. Methods: The experimental groups were divided into four groups: Normal, Joksamni(ST36), Joksamni(ST36)+Eumneungcheon(SP9), and Joksamni(ST36)+Gokji(LI11). Needles were inserted into acupoints at the depth of 0.5cm with basic insertion method. Electroacupuncture was done under the condition of 2Hz electrical biphasic pulses with continuous rectangular wave lasting for 0.2ms until the muscles produced visible contractions. Such stimulation was applied continuously for 10 minutes, 1 time every 2 days for 10 sessions of treatments. Thereafter we evaluated changes in NADPH-d positive neurons histochemically and changes in nNOS, NPY and VIP positive neurons immunohistochemically. Results: The optical densities of NADPH-d positive neurons of the Joksamni(ST36)+Eumneungcheon(SP9) group in all areas of cerebral cortex and Joksamni(ST36)+Gokji(LI11) group in primary somatosensory cortex, visual cortex, auditory cortex, perirhinal cortex were significantly increased as compared to the Joksamni(ST36) group. The optical densities of NADPH-d positive neurons of the Joksamni(ST36)+Gokji(LI11) group were significantly decreased as compared to the Joksamni(ST36)+Eumneungcheon(SP9) group with the exception of primary somatosensory cortex. The optical densities of nNOS positive neurons of the Joksamni(ST36)+Eumneungcheon(SP9) group in all areas of cerebral cortex and Joksamni(ST36)+Gokji(LI11) group in auditory cortex, perirhinal cortex, insular cortex were significantly increased as compared to the Joksamni(ST36) group. The optical densities of nNOS positive neurons of the Joksamni(ST36)+Gokji(LI11) group were significantly decreased in all areas of cerebral cortex as compared to the Joksamni(ST36)+Eumneungcheon(SP9) group. The optical densities of NPY positive neurons of the Joksamni(ST36)+Gokji(LI11) group were significantly decreased in primary motor cortex, primary somatosensory cortex, cingulate cortex as compared to the Joksamni (ST36) and Joksamni(ST36)+Eumneungcheon(SP9) groups. The optical densities of VIP positive neurons of the Joksamni(ST36)+Eumneungcheon(SP9) group were significantly increased in all areas of cerebral cortex except for cingulate cortex as compared to the Joksamni(ST36) group. The optical densities of VIP positive neurons of the Joksamni(ST36)+Gokji(LI11) group were significantly decreased in auditory cortex, cingulate cortex, perirhinal cortex as compared to the Joksamni(ST36) group. The optical densities of VIP positive neurons of the Joksamni(ST36)+Gokji(LI11) group were significantly decreased in all areas of cerebral cortex as compared to the Joksamni(ST36)+Eumneungcheon(SP9) group. Conclusions: The result demonstrated that electroacupuncture on Joksamni(ST36) and its combination change the activities of the NO system and peptidergic system in the cerebral cortex of SHR and that acupoint combination is one of the important parameters for the effects.

• The Korean Journal of Malacology
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• v.15 no.1
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• pp.31-39
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• 1999

This study was aimed to describe the population dynamics and secondary production of Tapes philppinarum on Chohwa and Toksan tidal flats in Kwangyang Bay, from June 1994 to July 1995. The size distributions in shell length showed bimodal patterns from June 1994 to April 1995. Thereafter the distribution changed to the passage of time, indicating an increase in shell length. The mean densities decreased gradually with month. Chohwa A sustained highestdensity, followed by Chohwa B, Toksan C, and Toksan D. The recruitment of new year class occurred first on the upper tidal flat, and then moved to the lower tidal flat. The mean biomass in flesh dry weight of each year class increased in spring just before the spawning period, and thereafter decreased progressively. Annual mean biomass wasrecorded as 170.7 g m$\^$-2/ at Chohwa A, 220.7 g m$\^$-2/ at Chohwa B, 21.8 g m$\^$-2/ at Toksan C, and 45.2 g m$\^$-2/ at toksan D. Annual productions of Ruditapes philippinarum were estimated as 259.72 g m$\^$-2/ yr$\_$-1/ at Chohwa A, 359.79 g m$\^$-2/ yr$\_$-1/ at Chohwa B, 45.02 g m$\^$-2/ yr$\_$-1/ at Toksan c, 68.88 g m$\^$-2/ yr$\_$-1/ at Toksan D. Annual productions were much higher on Chohwa tidal flat than those on Toksan tidal flat. While P:B ratio were higher on Toksan tidal flat as 1.70 than those on Chohwa tidal flat as 1.58. In conclusion, the Chohwa tidal flat showed higher biomass and producion of Ruditapes philippinarum, whereas the Toksan tidal flat showed higher P:B ratio. The total amount of potential food might influence on the density, biomass nad produciton, whereas the amount of food and space per individual might influence on the growth rate and P:B ratio.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.14 no.4
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• pp.212-216
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• 1981

The growth of Lactobacillus bulgaricus treated with vanillin, ortho-vanillin and guaiaco1 was studied on synthetic medium in mechanically agitated chemostat culture, The exponential-phase growth rate exhibited a maximum at the cells treated with 50 ppm vanillin. That stimulation, however, appears to be an effect on growth rate rather than total cell growth. And the others were inhibited by the chemicals. Much greater inhibition in growth of the cells treated with 100 ppm of each chemical than oars treated with 50 ppm was observed after 25 hour fomentation. For aerobic microbes, the alcohol dehydrogenase reaction is enhanced for the reproduction of NAD, which consequently cause to stimulate fermentation. For micro-aerophilic microbes , however, the same effect was not observed at the present study at least in the case of cell concentration. However except f or one treated with 50 ppm vanillin the same effect was observed in the case of growth is to. From the result using the glucose as a substrate, it was found that the cell concentrations measured in terms of ultimate optical density (UOB/ml), were 0.96 and 0.92, when treated with 50 and 100 ppm vanillin; 0.40 and 0.45 when treated with ortho-vanillin 50 and 100 ppm: 0.49 and 0.47, when treated with guaiacol 50 and 100 ppm. The specific growth rates were 0.44, 0.15, 0.25, 0.29, 0.37, and 0.34; the specific production rates wire 0.33, 0.15, 0.16, 0.22, 0.28, and 0.26 and the glucose concentrations (g/1) after 25 hour fermentation were 23.5, 32.8, 31.5, 29.5, 28.0 and 28.8, these all in the same sequences as the first.

• Proceedings of the Korean Biophysical Society Conference
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• 2003.06a
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• pp.80-80
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• 2003

${\beta}$-lapachone(${\beta}$-Lap), a natural o-naphthoquinone, presents in the bark of the Lapacho tree. ${\beta}$-Lap is cytotoxic against a variety of human cancer cells and it potentiates the anti-tumor effect of Taxol. In addition, ${\beta}$-Lap has been reported to radiosensitize cancer cells by inhibiting the repair of radiation-induced DNA damage.In the present study, we investigated the cytotoxicity of ${\beta}$-Lap against RKO human colorectal cancer cells as well as the combined effect of ${\beta}$-LaP and ionizing radiation. An incubation of RKO cells with 5 ${\mu}$M of ${\beta}$-Lap for 4 h killed almost 90％ of the clonogenic cells. An incubation of RKO cells with 5 ${\mu}$M of ${\beta}$-Lap for 4 h or longer also caused massive apoptosis. Unlike other cytotoxic agents, ${\beta}$-Lap did not increase the expression of p53 and p21 and it suppressed the NFkB expression. The expression of Caspase 9 and 3 was minimally altered by ${\beta}$-Lap. Radiation and ${\beta}$-Lap acted synergistically in inducing clonogenic cell death and apoptosis in RKO cells when ${\beta}$-Lap treatment was applied after but not before the radiation exposure of the cells. Interestingly, a 4 h treatment with 5 ${\mu}$M of ${\beta}$-Lap starting 5 h after irradiation was as effective as that starting immediately after irradiation. The mechanisms of ${\beta}$-Lap-induced cell killing is controversial but a recent hypothesis is that ${\beta}$-Lap is activated by NAD(P)H: quinone-onidoreductase (NQO1) in the cells followed by an elevation of cytosolic Ca$\^$2+/ level and activation of proteases leading to apoptosis. It has been reported that NQO1 level in cells is markedly up-regulated for longer than 10 h after irradiation. Indeed, using immunological staining of NQO1, we observed a significant elevation of NQO1 expression in RKO cells 5h after 2-4 Gy irradiation. Such a prolonged elevation of NQO1 level after irradiation may be the reasons why the ${\beta}$-Lap treatment applied S h after irradiation was as effective as that applied immediately after irradiation in killing the cells. In view of the fact that the repair of radiation-induced damage is usually completed within 1-2 h after irradiation, it is highly likely that the ${\beta}$-Lap treahment applied 5 h after irradiation could not inhibit the repair of radiation-induced damage. For in vivo study, RKO cells were injected S.C. into the hind-leg of Nu/Nu mice, and allowed to grow to 130 mm3 tumor. The mice were i.p. injected with ${\beta}$-lapachone or saline 2 h after irradiation of tumors with 10 Gy of X-rays. The radiation induced growth delay was increased by 2.4 $\mu\textrm{g}$/g of ${\beta}$-lapachone. Taken together, we may conclude that the synergistic interaction of radiation and ${\beta}$-Lap in killing cancer cells is not due to radiosensitization by ${\beta}$-Lap but to an enhancement of ${\beta}$-Lap cytotoxicity by radiation through an upregulation of NQO1. The fact that NQO1 is elevated in tumors and that radiation causes prolonged increase of the NQO1 expression may be exploited to preferentially kill tumor cells using ${\beta}$-Lap in combination with radiotherapy.