• Molecular & Cellular Toxicology
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• 제1권2호
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• pp.111-115
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• 2005

Ultraviolet (UV) radiation to mammalian skin is known to alter cellular function via generation of Reactive Oxygen Species (ROS), DNA damage and DNA lesions, such as pyrimidine dimmers and photoproducts, which could lead to DNA mutation if they are not repaired. In this study, we have investigated the reduction of DNA damage and of apoptosis with a particular attention to genetic effect of paeoniflorin in Normal Human Epidermal Keratinocytes (NHEK). After UVB irradiation from $10\;to\;500mJ/cm^{2}$ to NHEK, Mean Tail Moments (MTM) were increased with UVB dose increase. The greatest amount of strand breaks was induced at $500mJ/cm^{2}$ of UVB. Even at the lowest dose of UVB ($10mJ/cm^{2}$), change in MTM was detected (P<0.0001). Pretreated cell with 0.1% paeoniflorin maximally reduced the level of DNA damage to about 21.3%, compared to untreated cell. In the lower concentrations less than 0.01% of paeoniflorin, MTM had a small increase but paeoniflorin still had reductive effects of DNA damage. We measured the apoptosis suppression of paeoniflorin with annexin V flous staining kit. As we observed under the fluorescence microscopy to detect apoptosis in the irradiated cell, the fluorescence intensity was clearly increased in the untreated cell, but decreased in treated cells with paeoniflorin. These results suggest that paeoniflorin reduces the alteration of cell membranes and prevents DNA damage. Therefore, the use of paeoniflorin as a free radical scavenger to reduce the harmful effects of UV lights such as chronic skin damage, wrinkling and skin cancer can be useful to prevent the formation of photooxidants that result in radical damage.

• 동의생리병리학회지
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• 제26권4호
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• pp.483-490
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• 2012

This study was designed to identify the effects of Polygoni cuspidati Radix(PCR) on the generation of superoxide anion radicals (${\cdot}O_2{^-}$), nitric oxide (NO), peroxynitrite ($ONOO^-$) in the renal epithelial cells of mouse(LLC-$PK_1$). The effects of PCR on the expression of inflammation-related proteins, IKK-${\alpha}$, phospho-$I{\kappa}B-{\alpha}$, NF-${\kappa}B$ (p50, p65), COX-2, iNOS, IL-$1{\beta}$, VCAM-1, were examined by western blotting. For this study, the fluorescent probes, namely dihydrorhodamine 123 (DHR 123), 2',7'-dichloro dihydrofluorescein diacetate (DCFDA), 4,5-diaminofluorescein (DAF-2) were used. Protein expression levels of IKK-${\alpha}$, phospho-$I{\kappa}B-{\alpha}$, NF-${\kappa}B$ (p50, p65), COX-2, iNOS, IL-$1{\beta}$, VCAM-1 were assayed by western blot. PCR reduced $H_2O_2$-induced cell death dose-dependently. It inhibited the generation of ${\cdot}O_2{^-}$, NO, $ONOO^-$ and $PGE^2$ in the $H_2O_2$-treated LLC-PK1 cells in vitro. PCR inhibited the espression of IKK-${\alpha}$, phospho-$I{\kappa}B-{\alpha}$, COX-2, iNOS, IL-$1{\beta}$ and VCAM-1 genes by means of decreasing the NF-${\kappa}B$ activation. These results suggest that PCR is an effective NO, ${\cdot}O_2{^-}$, $ONOO^-$ scavenger, and this substance recommended to be applied in treatment for the inflammatory process and inflammation-related disease.

• 동의생리병리학회지
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• 제21권6호
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• pp.1499-1505
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• 2007

Peroxynitrite ($ONOO^-$) is a reactive oxidant formed from superoxide anion radical (${\cdot}\;O_2-$) and nitric oxide (NO), which can oxidize cellular components such as essential protein, non-protein thiols, DNA, low-density lipoproteins and membrane phospholipids. ${\cdot}\;O_2-$ and $ONOO^-$ have contributed to the pathogenesis of diseases such as stroke, heart disease, Alzheimer's disease and atherosclerosis. Because of damaging effects of ${\cdot}\;O_2-$ and $ONOO^-$ oxidants, Vespae Nidus, which has been known to strengthen the kidneys to preserve the vital energy. was tested as a potential specific scavenger of those oxidants. In this study, the viability of Vespae Nidus (1, 10, 50 g/ml) to scavenge ${\cdot}\;O_2-$, NO, $ONOO^-$ and so to protect cells against tert-butylhydroxyperoxide (t-BHP) induced cell death was tested. The levels of ${\cdot}\;O_2-$ and $ONOO^-$ were detected by staining with DCFH-DA and DHR 123, respectively. Protein expression levels of COX-2, iNOS and $NF{-\kappa}B$ were assayed by western blot. Vespae Nidus blocked t-BHP-induced cell death in a dose-dependent fashion. Vespae Nidus inhibited t-BHP-induced production of ${\cdot}\;O_2-$, NO and $ONOO^-$ in YPEN cells. The lipid peroxide level was increased and glutathione level was decreased in lipopolysaccharide (LPS)-treated ICR mouse, whereas the ones in the Vespae Nidus-administered group were regulated beneficially. Vespae Nidus inhibited the expression of COX-2, iNOS and NF-κB (p65 and p50) genes in LPS-treated ICR mouse. The present study suggests that Vespae Nidus is a powerful antioxidant and promotes cellular defense activity by scavenging the toxic oxidants such as ${\cdot}\;O_2-$ and $ONOO^-$.

• Radiation Oncology Journal
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• 제34권3호
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• pp.230-238
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• 2016

Purpose: Hypoxia can impair the therapeutic efficacy of radiotherapy (RT). Therefore, a new strategy is necessary for enhancing the response to RT. In this study, we investigated whether the combination of nanoparticles and RT is effective in eliminating the radioresistance of hypoxic tumors. Materials and Methods: Gold nanoparticles (GNPs) consisting of a silica core with a gold shell were used. CT26 colon cancer mouse model was developed to study whether the combination of RT and GNPs reduced hypoxia-induced radioresistance. Hypoxia inducible $factor-1{\alpha}$ ($HIF-1{\alpha}$) was used as a hypoxia marker. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining were conducted to evaluate cell death. Results: Hypoxic tumor cells had an impaired response to RT. GNPs combined with RT enhanced anti-tumor effect in hypoxic tumor compared with RT alone. The combination of GNPs and RT decreased tumor cell viability compare to RT alone in vitro. Under hypoxia, tumors treated with GNPs + RT showed a higher response than that shown by tumors treated with RT alone. When a reactive oxygen species (ROS) scavenger was added, the enhanced antitumor effect of GNPs + RT was diminished. Conclusion: In the present study, hypoxic tumors treated with GNPs + RT showed favorable responses, which might be attributable to the ROS production induced by GNPs + RT. Taken together, GNPs combined with RT seems to be potential modality for enhancing the response to RT in hypoxic tumors.

• 미생물학회지
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• 제50권1호
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• pp.22-26
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• 2014

Protocatechualdehyde (PA)는 항산화 활성과 항암 활성을 가진 페놀성 물질이다. Streptomyces lincolnensis M-20 균주에서 생산된 PA를 균주 상등액에서 분리, 정제하였다. 항산화 활성을 가진 PA가 구리 이온 존재 하에서는 산화촉진제로 작용하였다. 항산화 활성은 DPPH를 이용한 방법으로 측정하였으며, 구리 이온 존재 하에서 PA의 산화 촉진 작용은 pBR322 플라스미드의 DNA 절단 작용으로 측정하였다. DNA 손상으로 생성되는 활성산소 종의 확인은 활성 산소종의 포집자인 글루타치온에 의해 DNA 절단이 억제되는 것으로 확인하였다. PA와 구리 이온의 복합체 형성은 금속 이온의 킬레이트인 EDTA가 존재할 경우와 존재하지 않을 경우를 자외선/가시광선 분광학적 분석법으로 비교, 확인하였다.

• The Korean Journal of Physiology and Pharmacology
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• 제11권3호
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• pp.85-88
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• 2007

Matrix metalloproteinases (MMPs) can degrade a wide range of extracellular matrix components. It has been reported that MMP-9 are activated after focal ischemia in experimental animals. (-)-Epigallocatechin-3-gallate (EGCG), a major constituent of green tea polyphenols, is a potent free radical scavenger and reduces the neuronal damage caused by oxygen free radicals. And it has been known that EGCG could reduce the infarction volume in focal brain ischemia and inhibit MMP-9 activity. To delineate the relationship between the anti-ischemic action and the MMP-9-inhibiting action of EGCG, we investigated the effect of EGCG on brain infarction and the activity of matrix metalloproteinase-9 induced by permanent middle cerebral artery occlusion (pMCAO) in ICR mice. EGCG (40 mg/kg, i.p. $15{\sim}30min$ prior to MCAO) significantly decreased infarction volume at 24 hr after MCAO. GM 6001 (50 mg/kg, i.p. $15{\sim}30min$ prior to MCAO), a MMP inhibitor, also significantly reduced infarction volume. In zymogram, MMP-9 activities began to increase at ipsilateral cortex at 2 hr after MCAO, and the increments of MMP-9 activities were attenuated by EGCG treatment. Western blot for MMP-9 also showed patterns similar to that of zymogram. These findings demonstrate that the anti-ischemic action of EGCG ire mouse focal cerebral ischemia involves its inhibitory effect on MMP-9.

• Archives of Pharmacal Research
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• 제29권2호
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• pp.145-151
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• 2006

We investigated the effects of Ginsenoside $R_e$ on human sperm motility in fertile and asthenozoospermic infertile individuals in vitro and the mechanism by which the Ginsenosides play their roles. The semen samples were obtained from 10 fertile volunteers and 10 asthenozoospermic infertile patients. Spermatozoa were separated by Percoll and incubated with 0, 1, 10 or $100\;{\mu}M$ of Ginsenoside $R_e$. Total sperm motility and progressive motility were measured by computer-aided sperm analyzer (CASA). Nitric oxide synthase (NOS) activity was determined by the $^{3}H$-arginine to $^{3}H$-citrulline conversion assay, and the NOS protein was examined by the Western blot analysis. The production of sperm nitric oxide (NO) was detected using the Griess reaction. The results showed that Ginsenoside $R_e$ significantly enhanced both fertile and infertile sperm motility, NOS activity and NO production in a concentration-dependent manner. Sodium nitroprusside (SNP, 100 nM), a NO donor, mimicked the effects of Ginsenoside $R_e$. And pretreatment with a NOS inhibitor $N^{w}$-Nitro-L-arginine methyl ester (L-NAME, $100\;{\mu}M$) or a NO scavenger N-Acetyl-L-cysteine (LNAC, 1 mM) completely blocked the effects of Ginsenoside $R_e$. Data suggested that Ginsenoside $R_e$ is beneficial to sperm motility, and that induction of NOS to increase NO production may be involved in this benefit.

• 한국수정란이식학회지
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• 제30권1호
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• pp.23-31
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• 2015

Catalpol, an iridoid glucoside, isolated from the root of Rehmannia glutinosa Libosch. It possesses a broad range of biological and pharmacological activity including anti-tumor, anti-inflammation and anti-oxidant by acting as a free radical scavenger. Therefore, in this study, the effects of catalpol on blastocyst development, expression levels of reactive oxygen species (ROS) and apoptotic index were investigated in porcine embryos. After in vitro maturation and fertilization, porcine embryos were cultured for 6 days in porcine zygote medium 3 (PZM-3) supplemented with catalpol (0, 100, 200 and $400{\mu}M$, respectively). Blastocyst development not significantly improved in the catalpol treated group when compared with control group. Otherwise, the intracelluar levels of ROS were decreased and the numbers of apoptotic nuclei were reduced in the catalpol ($100{\mu}M$) treated porcine blastocysts (P<0.05). On the other hand, blastocyst development was significantly improved in the catalpol ($100{\mu}M$) treated group when compared with the untreated catalpol group under $H_2O_2$ ($200{\mu}M$) induced oxidative stress (P<0.05). Otherwise, the intracellular levels of ROS in catalpol ($100{\mu}M$) treated group were significantly decreased in the untreated catalpol group under $H_2O_2$ ($200{\mu}M$) induced oxidative stress (P<0.05). Furthermore, the total cell numbers of blastocysts were significantly increased (P<0.05) in the catalpol ($100{\mu}M$) treated group under $H_2O_2$ ($200{\mu}M$) induced oxidative stress, whereas numbers of apoptoic nuclei were significantly reduced (P<0.05). In conclusion, our results indicate that treatment of catalpol may have important implications for improving developmental competence and preimplantation quality of porcine embryos through its anti-oxidant and anti-apoptotic effect.

• 대한한방내과학회지
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• 제27권3호
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• pp.688-697
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• 2006

Objectives : Peroxynitrite ($ONOO^-$), $O_2^-$ and nitric oxide (NO) are cytotoxic species that can oxidize several cellular components such as proteins. lipids and DNA. It has been implicated in the aging process and age-related disease such as Alzheimer's disease, rheumatoid arthritis, cancer and atherosclerosis. The aim of this study was to investigate $ONOO^-$ scavenging activities. and that of its precursors. NO and $O_2^-$ of Trigonellae Semen. Methods : To investigate $ONOO^-$. NO. $O_2^-$ scavenging activities, fluorescent probes, namely 2'.7'-dichlorodihydrofluorescein diacetate (DCFDA). 4.5-diaminofluorescein (DAF-2) and dihydrorhodamine 123 (DHR 123) were used. Protein expression levels of iNOS, COX-2, NF-${\kappa}B$, and VCAM-1 were assayed by western blot. Results : Trigonellae Semen markedly scavenged authentic $ONOO^-$, $O_2^-$ and NO. It also inhibited $ONOO^-$ induced by $O_2^-$ and NO which are derived from SIN-1. Furthermore. Trigonellae Semen inhibited $ONOO^-$, $O_2^-$ and NO generation in LPS-treated ICR mouse kidney postmitochondria. Trigonellae Semen inhibited gene expression of iNOS, COX-2, VCAM-l and NF-${\kappa}B$ (p65) activation. Conclusions : These results suggest that Trigonellae Semen is an effective $ONOO^-$, $O_2^-$ and NO scavenger. and that this substance has a potential role as an inhibitor of the aging process, and in therapy against age-related diseases.

• Biomolecules & Therapeutics
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• 제25권3호
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• pp.308-314
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• 2017

Urotensin II (UII) is a mitogenic and hypertrophic agent that can induce the proliferation of vascular cells. UII inhibition has been considered as beneficial strategy for atherosclerosis and restenosis. However, currently there is no therapeutics clinically available for atherosclerosis or restenosis. In this study, we evaluated the effects of a newly synthesized UII receptor (UT) antagonist, KR-36996, on the proliferation of SMCs in vitro and neointima formation in vivo in comparison with GSK-1440115, a known potent UT antagonist. In primary human aortic SMCs (HASMCs), UII (50 nM) induced proliferation was significantly inhibited by KR-36996 at 1, 10, and 100 nM which showed greater potency ($IC_{50}$: 3.5 nM) than GSK-1440115 ($IC_{50}$: 82.3 nM). UII-induced proliferation of HASMC cells was inhibited by U0126, an ERK1/2 inhibitor, but not by SP600125 (inhibitor of JNK) or SB202190 (inhibitor of p38 MAPK). UII increased the phosphorylation level of ERK1/2. Such increase was significantly inhibited by KR-36996. UII-induced proliferation was also inhibited by trolox, a scavenger for reactive oxygen species (ROS). UII-induced ROS generation was also decreased by KR-36996 treatment. In a carotid artery ligation mouse model, intimal thickening was dramatically suppressed by oral treatment with KR-36996 (30 mg/kg) which showed better efficacy than GSK-1440115. These results suggest that KR-36996 is a better candidate than GSK-1440115 in preventing vascular proliferation in the pathogenesis of atherosclerosis and restenosis.