• Molecules and Cells
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• v.45 no.3
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• pp.134-147
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• 2022

The anti-oxidant enzyme heme oxygenase-1 (HO-1) is known to exert anti-inflammatory effects. From a library of pyrazolo[3,4-d]pyrimidines, we identified a novel compound KKC080096 that upregulated HO-1 at the mRNA and protein levels in microglial BV-2 cells. KKC080096 exhibited anti-inflammatory effects via suppressing nitric oxide, interleukin1β (IL-1β), and iNOS production in lipopolysaccharide (LPS)-challenged cells. It inhibited the phosphorylation of IKK and MAP kinases (p38, JNK, ERK), which trigger inflammatory signaling, and whose activities are inhibited by HO-1. Further, KKC080096 upregulated anti-inflammatory marker (Arg1, YM1, CD206, IL-10, transforming growth factor-β [TGF-β]) expression. In 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridinetreated mice, KKC080096 lowered microglial activation, protected the nigral dopaminergic neurons, and nigral damage-associated motor deficits. Next, we elucidated the mechanisms by which KKC080096 upregulated HO-1. KKC080096 induced the phosphorylation of AMPK and its known upstream kinases LKB1 and CaMKKbeta, and pharmacological inhibition of AMPK activity reduced the effects of KKC080096 on HO-1 expression and LPS-induced NO generation, suggesting that KKC080096-induced HO-1 upregulation involves LKB1/AMPK and CaMKKbeta/AMPK pathway activation. Further, KKC080096 caused an increase in cellular Nrf2 level, bound to Keap1 (Nrf2 inhibitor protein) with high affinity, and blocked Keap1-Nrf2 interaction. This Nrf2 activation resulted in concurrent induction of HO-1 and other Nrf2-targeted antioxidant enzymes in BV-2 and in dopaminergic CATH.a cells. These results indicate that KKC080096 is a potential therapeutic for oxidative stress-and inflammation-related neurodegenerative disorders such as Parkinson's disease.

• The Korean Journal of Food And Nutrition
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• v.21 no.4
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• pp.397-402
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• 2008

Over the past few years, anthocyanin has been demonstrated to exert potentially anti-oxidative and bioavailability effects including anti-cardiovascular disease and cancer-preventive effects in humans. This study was conducted to assess the toxicity of anthocyanin extracts from black soybean seed coats via oral administration in a mice feeding test. The final anthocyanin-containing products induced signs of toxicity on mortality in the 4-week feeding test. The incidence of clinical signs and changes in body and organ weight were also not observed in all anthocyanin-treated groups as compared with the control groups. In hematology analysis, RBC indices concluding MCV, MCH, and MCHC and WBC differential counting such a NEU, LYM, MONO, EOS, and BASO evidenced no significant differences between the anthocyanin treatment and control groups. The biochemical parameters in serum analysis - ALT, AST, BUN, Crea etc.- were also not significantly altered in those groups. Absolute and relative organ weights were not increased after 4 weeks of treatment with anthocyanin extracts in mice. In conclusion, anthocyanin extracts from the black soybean has biological activity without any toxic effects, which also suggests that the consumption of soy containing anthocyanin products may be a good source for health and medical supplies.

• Molecules and Cells
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• v.43 no.11
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• pp.964-973
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• 2020

Recent studies have highlighted that early enhancement of the glycolytic pathway is a mode of maintaining the proinflammatory status of immune cells. Thiamine, a wellknown co-activator of pyruvate dehydrogenase complex, a gatekeeping enzyme, shifts energy utilization of glucose from glycolysis to oxidative phosphorylation. Thus, we hypothesized that thiamine may modulate inflammation by alleviating metabolic shifts during immune cell activation. First, using allithiamine, which showed the most potent anti-inflammatory capacity among thiamine derivatives, we confirmed the inhibitory effects of allithiamine on the lipopolysaccharide (LPS)-induced pro-inflammatory cytokine production and maturation process in dendritic cells. We applied the LPS-induced sepsis model to examine whether allithiamine has a protective role in hyper-inflammatory status. We observed that allithiamine attenuated tissue damage and organ dysfunction during endotoxemia, even when the treatment was given after the early cytokine release. We assessed the changes in glucose metabolites during LPS-induced dendritic cell activation and found that allithiamine significantly inhibited glucose-driven citrate accumulation. We then examined the clinical implication of regulating metabolites during sepsis by performing a tail bleeding assay upon allithiamine treatment, which expands its capacity to hamper the coagulation process. Finally, we confirmed that the role of allithiamine in metabolic regulation is critical in exerting anti-inflammatory action by demonstrating its inhibitory effect upon mitochondrial citrate transporter activity. In conclusion, thiamine could be used as an alternative approach for controlling the immune response in patients with sepsis.

• Journal of Physiology & Pathology in Korean Medicine
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• v.27 no.6
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• pp.745-751
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• 2013

Safflower (Carthamus tinctorius L.) seeds have been used in Korea and China for promoting bone formation and protection. This study was designed to examine the Nrf-2 mediated anti-oxidative effects of Korean and Chinese safflower seeds. Water and ethanol extracts of safflower seeds were treated to RAW 264.7 cells. Nrf-2 transcriptional activity was measured by reporter gene assay and western blot analysis. Semi-quantitive RT-PCR analysis was adopted to measure Nrf-2 dependent gene expressions. Water extracts of safflower seeds have strongly induced the activation of Nrf-2 transcription than ethanol extracts. Especially, water extracts of Korean safflower seeds has more strongly increased the expression of nuclear Nrf-2. Water extracts of Korea and China safflower seeds have also increased the expression of Nrf-2-dependent genes such as GCLC, NQO-1 and HO-1 in RAW 264.7 cells. However, all kinds of safflower seeds extracts did not increase intracellular ROS production. These results demonstrate that the antioxidant effects of safflower seeds are not related with ROS production, rather it is mediated by the direct activation of Nrf-2.

• Toxicological Research
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• v.17
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• pp.271-280
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• 2001

We report on the applicability oj a method recently developed in our laboratory for measuring the antioxidant potential of isolated chemicals and extracts derived from natural products. Peroxyl radicals generated by thermal homolysis of 2,2'-azobis-amidinopropane (ABAP) oxidize $\alpha$-keto-${\gamma}$-methiolbutyric acid (KMBA) to ethylene, which is monitored by gas chromatography. Inhibition of ethylene formation in the presence of antioxidants that compete with KMBA for peroxyl radicals is the basis of the Total Oxyradical Scavenging Capacity Assay (TOSCA; Winston et al., 1998). Antioxidative activities of water-soluble extracts of American green tea, the anti-hepatotoxic flavonolignan from milk thistle (Silybum marianum) silybin, Ginkgo biloba extract EGb 761, and a series of naturally occuring sesquiterpene lactones (all ger-macranolides found in in fungi, liverworts, and plants) were studied. The specific TOSC value per $\mu$M silybin was 5.2, which is essentially comparable to that of Trolo $x^{ⓡ}$, a water-soluble vitamine E analog. Tea and Ginkgo extracts exhibited potent peroxyl radical scavenging capacity with values, respectively of =1700 and 1000 $\mu$mols Trolo $x^{ⓡ}$ equivalent per gram dry matter. The known anti-inflammatory activity of some germacranolides prompted study of their antioxidant capacity. None of the lactones exhibited antioxidant capacity toward peroxyl radicals comparable to Trolo $x^{ⓡ}$; costunilide, the most lipophilic, had a TOSC value = to glutathione. The potential role of peroxyl radicals in lipidperoxidation, other cellular damage, and var-ious disease states suggest a possible preventive role for silybin, green tea and Ginkgo biloba in oxidative stress caused by these free radical species.ecies.

• Toxicological Research
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• v.22 no.3
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• pp.229-235
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• 2006

Opuntia humifusa is a member of the Cactaceae family. In the present study, the antioxidant, nitric oxide(NO) inhibitory activities and potential irritation response of the fermented Opuntia humifusa cladodes(FOH) were investigated for cosmetic use. Antioxidant activities were tested using 1,1-diphenyl-2-picrylhydrazyl(DPPH) and xanthine oxidase assay, we found that FOH could scavenge DPPH free radicals and inhibit xanthine oxidase activity in a dose dependent manner, with $IC_{50}$ of 2599.46${\mu}g/ml$ and 721.38${\mu}g/ml$, respectively. To investigate the possible anti-inflammatory effects of FOH, RAW 264.7 macrophages were pretreated with FOH($0{\sim}400{\mu}g/ml$) for 30 min and then treated with LPS for 24 h. We found that cell number did not vary significantly with the treatment of FOH, and FOH did not show any inhibitory effects on LPS-induced NO production. After application of FOH to rabbits for skin and eye irritation test, the experimental sites did not show any response compared to the control. FOH were considered to be a non-irritant to the skin and eye. Based on the above information, we suggest that FOH can be considered to be a non-irritant base cosmetic material for safely use.

• Journal of Ginseng Research
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• v.35 no.4
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• pp.429-435
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• 2011

Korean red ginseng has been studied various biological activities such as immune, anti-oxidative, anti-microbial, and anticancer activities but antiviral mechanism needs further studies. In this study, we aimed to examine the antiviral effects of Korea red ginseng extract and ginsenosides on norovirus surrogate, including murine norovirus (MNV) and feline calicivirus (FCV). We evaluated the pre-, co-, and post-treatment effects of Korean red ginseng (KRG), ginsenosides $Rb_1$ and $Rg_1$. To measure the antiviral effect and cytotoxicity of KRG extract, and ginsenosides $Rb_1$ and $Rg_1$, we treated Crandell-Reese Feline Kidney for FCV or RAW264.7 cells for MNV with concentrations of 0, 5, 6.7, 10, 20 ug/mL total saponin. There was cytotoxic effect in the highest concentration 20 ug/mL of KRG extract so this concentration was excluded in this study. The FCV titer was significantly reduced to 0.23-0.83 $log_{10}$ 50% tissue culture infectious dose ($TCID_{50}$)/mL in groups pre-treated with red ginseng extract or ginsenosides. The titer of MNV was significantly reduced to 0.37-1.48 $log_{10}$ $TCID_{50}$/mL in groups pre-treated with red ginseng extract or ginsenosides. However, there was no observed antiviral effect in groups co-treated or post-treated with KRG and its constituents. Our data suggest that KRG extract has an antiviral effect against norovirus surrogates. The antiviral mechanisms of KRG and ginsenosides should be addressed in future studies.

• Korean Journal of Medicinal Crop Science
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• v.17 no.1
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• pp.68-74
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• 2009

Vitis amurensis (VA; Vitaceae) has long been used in oriental herbal medicine. It has been reported that roots and seeds of VA have anti-inflammatory and antioxidant effects. In the present study, the protective effect of ethanol extract from stems and leaves of VA on hydrogen peroxide (${H_2}{O_2}$) (100 ${\mu}M$)-induced neuronal cell damage was examined in primary cultured rat cortical neurons. VA (10-100 ${\mu}g$/ml) concentration-dependently inhibited ${H_2}{O_2}$-induced apoptotic neuronal cell death measured by 3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay and Hoechst 33342 staining. VA inhibited ${H_2}{O_2}$-induced elevation of intracellular $Ca^{2+}$ concentration (${[Ca^{2+}]}_i$) and generation of reactive oxygen species (ROS), which were measured by fluorescent dyes. Pretreatment of VA also prevented glutamate release into medium induced by 100 ${\mu}M$ ${H_2}{O_2}$, which was measured by HPLC. These results suggest that VA showed a neuroprotective effect on ${H_2}{O_2}$-induced neuronal cell death by interfering with ${H_2}{O_2}$-induced elevation of ${[Ca^{2+}]}_i$, glutamate release, and ROS generation. This has a significant meaning of finding a new pharmacological activity of stems and leaves of VA in the CNS.

• Environmental Mutagens and Carcinogens
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• v.20 no.1
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• pp.14-20
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• 2000

DHAQ-97, (2-(3-[p-bis(2-chloroethyl)aminophenyl]-2 formylaminopropanoyloxy) methy1-1,4-dihy-droxy-9,10-anthraquinone), is a novel anthraquinone derivative synthesized for use as an anti-neoplastic agent. In the present study, we have evaluated the selective cytotoxicity of DHAQ-97 by comparing its effects on viability and proliferation of human breast cancer cell line (NCF-7) versus normal immortalized breast epithelial cell line (MCF-10A). Thus, DHAQ-97 reduced both viability and proliferation of MCF-7 cells to a much greater extent than did for MCF-10A cells. The growth inhibitory and anti-proliferative properties of DHAQ-97 appear to be attributable to its ability to induce apoptosis as revealed by positive staining after in 냐셔 nick-end labeling (TUNEL), cleavage of poly(ADP-ribose)polymerase, release of mitochondrial cytochrome c into cytoplasm, and increased expression of pro-apoptotic Bax protein. Recent studies have indicated possible involvement of the ubiquitous eukaryotic transcription factor, NF-kappa B (NF-kB) in the regulation of apoptotic cell death. In line induced cytotoxicity in cultured MCF-7 cells. Furthermore, mild activation of NF-kB, as determined by its increased DNA binding capability, was observed 30 min after treatment with 10$\mu\textrm{m}$ DHAQ-97. Taken together, the above findings suggest that DHAQ-97 exerts selective cytotoxicity towards cancer cells through induction of apoptosis, which appears to be regulated by NF-kB.

• Biomedical Science Letters
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• v.24 no.3
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• pp.239-244
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• 2018

Recently, many researches regarding the natural products which alternate with chemical products have been done. Among them, boswellia is well known for effect on anti-oxidative effect and inflammation. The aim was the effect boswellia of formalin- induced orofacial and temporomandibular joint (TMJ) pain on experimental animals was investigated. Experiments were carried out using subcutaneous (SC) pain model and TMJ pain model that were induced by the injection of 5% formalin into the right vibrissa pad (SC, $50{\mu}L$) or TMJ ($30{\mu}L$) of rats, respectively. In both models, formalin (5%), formalin after distilled water (vehicle), formalin after boswellia extract (p.o., concentrations of 15, 30 mg/kg) (n=6). The number of scratching on the injected region was scored during the 9 successive periods of 5 min intervals following injection of formalin. Oral administration of boswellia (15, 30 mg / kg) reduced formalin-induced SC orofacial pain behavioral responses. SC orofacial pain behavioral responses was significantly reduced at 20~35 min. In the experimental group injected into temporomandibular joints, the pain response was significantly reduced by $276.2{\pm}8.20$ and $78.3{\pm}4.7$ after oral administration of boswellia (15, 30 mg / kg) at $398.3{\pm}24.8$ times. As a result of the passage of time, the oral administration of boswellia showed a significant effect of reducing the temporomandibular joint pain 30 minutes after the injection of formalin. This study confirmed that oral administration of boswellia modulated the pain behavior in both models. In conclusion, boswellia extract may be a potential therapeutic treatment for orofacial pain.