• 생명과학회지
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• 제19권5호
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• pp.607-614
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• 2009

본 연구는 말똥성게에 대한 항 발암 효과와 항산화 효과에 대하여 조사하였다. 먼저 말똥성게 분획물의 항 발암 효과를 알아보기 위하여 핵산, 메탄올, 부탄올 및 물로 순차적으로 분획하여 각 분획별로 암세포에 대한 증식 억제효과와 암 예방 지표인 QR 유도 활성 효과를 측정하였다. 그 결과 4종의 모든 암세포주 HepG2, MCF-7, HT29 및 B16-F10에서 농도 의존적인 암세포 증식 억제 효과를 나타내었으며, 모두 HPMH층에서 가장 높은 암세포 성장 억제 효과를 나타내었다. 그러나 HepG2에서 측정한 QR 유도 활성 효과는 HPMB층에서 가장 높은 효과를 보였다. 또한 항산화 효과 중 ROS 제거활성 측정결과에서는 HPMH층이 가장 높게 나타났으며 13.00 ${\mu}g/ml$에서 50%의 ROS 제거활성 효과를 나타내어 positive control로 사용한 trolox와 비교하였을 때 거의 유사한 제거활성을 보였다. 그리고 $ONOO^-$ 제거활성 역시 HPMH층이 가장 높게 나타났으며, positive control과 비교하였을 때 penicillamine의 $IC_{50}$값인 5.51 ${\mu}g/ml$ 보다는 낮았으나 15.14 ${\mu}g/ml$의 $IC_{50}$값을 나타내어 다른 분획층에 비해서는 높은 항산화 효과를 확인할 수 있었다. 아질산염 제거활성 측정 결과에서도 HPMH층이 가장 높은 제거활성 효과를 나타내었으며 특히 pH가 낮을수록, 시료농도가 증가할수록 아질산염 제거활성 효과가 증가함을 알 수 있었다. 이상으로 항 발암효과 중 암세포에 대한 증식 억제효과는 HPMH층이 가장 높았고 QR 유도 활성 효과는 HPMB층에서 가장 높았으며 항산화 효과는 ROS, ONOO- 및 아질산염 제거활성에서 모두 HPMH층이 가장 높음을 알 수 있었다. 따라서 앞으로 HPMH층과 HPMB층에 대한 집중적인 연구가 요구되어지며 말똥성게를 이용하여 항암관련 기능성 식품과 식품 저장성과 안전성 향상을 위한 식품첨가제나 보존제등의 개발가능성과 그 효과도 기대할 수 있을 것으로 생각된다.

• 동의생리병리학회지
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• 제30권4호
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• pp.242-249
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• 2016

This study was to investigate the anti-inflammatory effects of Prunellae Herba(PH). The generation of superoxide anion radical (․O₂-), nitric oxide (NO), peroxynitrite (ONOO-) and Prostaglandin E₂(PGE₂) were measured in the H₂O₂-Treated renal epithelial cells(LLC-PK1 cell) of mouse. And the effects of Prunellae Spica on the expression of NF-κB (p50, p65), IKK-α, phospho-IκB-α and inflammation-related proteins, COX-2, iNOS, IL-1β and VCAM-1, were examined by western blot. The fluorescent probes, 2',7'-dichlorodihydrofluorescein diacetate (DCFDA), 4,5-diaminofluorescein (DAF-2) and dihyldrorhodamine 123 (DHR 123) were used to estimate the scavenging effect of Prunellae Spica on ․O₂-, NO, ONOO-. Western blot was conducted to assess the protein expression levels of NF-κB (p50, p65), IKK-α, phospho-IκB-α, inflammation-related proteins, COX-2, iNOS, IL-1β, VCAM-1. PH inhibited H₂O₂-treated cell death dose-dependently. It reduced the generation of ·O₂-, NO, ONOO- and PGE₂ in the H₂O₂-treated renal epitheial cells(LLC-PK₁ cell) of mouse in vitro. PH reduced the expression of NF-κB, IKK-α, phospho-IκB-α, COX-2, iNOS, IL-1β and VCAM-1 genes through means of decreasing activation of NF-κB signaling as well. According to these results, PH has an antioxidative activity and anti-inflammatory effect by regulating the NF-κB pathway. This suggest that PH is expected to be used to regulating inflammatory process and treating inflammation-related disease.

• 농업과학연구
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• 제42권4호
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• pp.407-414
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• 2015

본 실험에서는 청국장의 항산화 기능성 증진을 위해 녹차를 첨가한 서리태 청국장을 제조한 후 LLC-$PK_1$ 세포를 이용하여 SIN-1유도 세포 독성에 대한 보호효과를 검토하였다. 항산화능이 대두보다 우수한 서리태를 주재료로, 부재료로 녹차를 농도별(0.5%, 2.0%, 5.0%)로 달리하여 청국장을 각각 제조한 후 $ONOO^-$에 의해 유도된 산화적 스트레스에 대해 서리태 청국장과 녹차첨가 서리태 청국장의 산화스트레스에 대한 개선효과를 비교한 결과, 녹차첨가 서리태청국장 추출물은 SOD, GSH-Px와 같은 항산화효소의 활성을 증가시키고, 지질과산화를 억제하여 세포생존율을 증가시킴으로써 산화적 스트레스에 대한 보호효과를 나타냈었으며, 특히 녹차를 5.0% 첨가했을 때의 효과가 가장 높은 것으로 나타났다. 따라서, 녹차 첨가 서리태 청국장은$ONOO^-$에 의해 유발된 산화적 스트레스를 개선함으로써 이로 인한 질병을 예방하는 항산화제로서 역할을 할 것으로 생각된다

• 대한한방내과학회지
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• 제31권1호
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• pp.153-165
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• 2010

Objectives : The aims of this study were to investigate the cytoprotective, antioxidative and inflammation genes inhibitory effects of Patriniae Radix on the mouse LLC-$PK_1$ cells (renal epithelial cells). Methods : The cytoprotective effect of Patriniae Radix was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The antioxidative effect was measured in terms of generation amount of superoxide anion radical (${\cdot}{O_2}^-$) by 2',7'-dichlorodihydrofluorescein diacetate (DCFDA), nitric oxide (NO) by 4,5-diaminofluorescein (DAF-2), peroxynitrite ($ONOO^-$) by dihyldrorhodamine 123 (DHR 123) and prostaglandin $E_2$ ($PGE_2$) by $PGE_2$ immunoassay on $H_2O_2$-treated LLC-$PK_1$ cells. For measuring of inflammation genes inhibitory effects, western blot was performed to detect IKK-$\alpha$, phospho-$I{\kappa}B-\alpha$, NF-${\kappa}B$ (p50, p65), COX-2, iNOS, IL-$1{\beta}$ and VCAM-1 protein level in cytosol fractions from LLC-$PK_1$ cells. Results : Patriniae Radix extract reduced the $H_2O_2$-induced cell death and inhibited the amount of $H_2O_2$-induced ${\cdot}{O_2}^-$, NO, $ONOO^-$, $PGE_2$ generation dose-dependently on the mouse LLC-$PK_1$ cells in vitro. Also Patriniae Radix extract inhibited the expression of IKK-$\alpha$, phospho-$I{\kappa}B-\alpha$, COX-2, iNOS, IL-$1\beta$ and VCAM-1 genes dose-dependently by means of decreasing activation of NF-${\kappa}B$. Conclusions : According to above results, it was identified that Patriniae Radix had the cytoprotective, antioxidative and inflammation genes inhibitory effects. So it was suggested that Patriniae Radix would be effective to the treatment for the inflammatory process and inflammation-related diseases.

• 생명과학회지
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• 제19권1호
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• pp.52-57
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• 2009

본 연구는 천연소재용 이용하여 식품 저장성과 안전성 향상을 위한 식품첨가제나 보존제등의 개발 가능성을 알아보고자 민간에서 사용되어 온 노박덩굴 분획물에 대한 항산화 효과 및 항균활성을 측정하였다. DPPH free radical 소거활성을 측정하여 항산화 활성효과를 알아 본 결과 COMB층이 $39.75{\mu}g/ml$에서 50%의 DPPH free radical의 소거활성을 보여 가장 높은 항산화 효과를 나타내었다. ROS 제거활성 측정결과에서도 COMB층이 가장 높은 항산화 효과를 나타내었으며 $4.73{\mu}g/ml$에서 50%의 ROS 제거 활성을 나타내어 positive control로 사용한 trolox와 거의 유사한 제거활성을 보였다. 그리고 $ONOO^-$ 제거활성 역시 COMB층이 가장 높게 나타났으며, 특히 positive control인 penicillamine 의 IC50값인 $4.34{\mu}g/ml$과 비교하였을 때 더 높은 제거 활성 효과를 나타내어 높은 항산화 효과를 확인할 수 있었다. 또한 paper disc method를 이용한 항균효과 실험 결과, 사용한 모든 균주에서 COMH층을 제외한 COMM, COMB 및 COMA층이 항균활성을 나타내었다. 특히 bacillus subtilis에서는 COMH층을 제외한 모든 층에서 농도에 비례하여 항균력이 점차 증가하였으며, 전반적으로 COMM층과 COMB층에서 높은 항균활성을 나타냄을 알 수 있었다. 본 연구 결과를 종합해보면 항산화효과는 노박덩굴의 butanol 분획층인 COMB층에서 가장 높았으며 그 다음으로 COMM 순이었다. 항균 효과는 사용한 모든 균주에서 methanol 분획층인 COMM 층이 전반적으로 높은 항균활성이 나타났으며 COMB층이 다음으로 높은 항균활성을 나타내었다. 이 상의 결과로 노박덩굴의 butanol 분획층인 COMB층과 methanol 분획층인 COMM 중에서 항산화 활성과 항균력을 나타내는 효과적인 생리 활성물질이 함유되어 있을 것으로 사료되어 지며, 앞으로 더욱 심도 있는 연구용 통하여 천연 항산화제, 천연식품보조제 및 천연 항균제등의 기능성 식품으로서의 활용가능성을 기대할 수 있을 것으로 생각된다.

• 대한한의학회지
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• 제28권1호통권69호
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• pp.237-248
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• 2007

Objectives : Diabetes is a disease in which the body does not produce or properly use insulin. Etiological studies of diabetes and its complications showed that oxidative stress might play a major role. Therefore, many efforts have been tried to regulate free oxygen radicals for treating diabetes and its complications. Iksujisundan has been known to be effective for the treatment of diabetes. The present study was carried out to investigate the effect of Iksujisundan on renal function, peroxynitrite(ONOO-) scavenging activity and polyol pathway in streptozotocin-induced diabetic rats. Methods : The crushed Iksujisundan was extracted 3 times, each time with 3 volumes of methyl alcohol at 60$^{\circ}C$ for 24 h. The extract was filtered and evaporated under a reduced pressure using a rotary evaporator to yield 87.8g. Iksujisundan extract was orally administreted at 100 mg per 1 kg of body weight for 20 days to the diabetic rats induced by streptozotocin(60mg/kg). The effects of Iksujisundan extract on the streptozotocin-induced diabetic rats were observed by measuring the serum level of glucose, insulin, lipid components, creatinine and BUN, and also the kidney levels of superoxide anion radical(${\cdot}$O2-), nitric oxide(NO) and ONOO-, and also the enzyme activities involved in the polyol pathway. Results : The effects of Iksujisundan on the streptozotocin-induced diabetic rats with regards to body weight, blood glucose and indulin levels, creatinine and BUN levels, total cholesterol and triglyceride lavels, and HDL-cholesterol levels were all shown to be good enough to prevent and cure the diabetes and its complications. Iksujisundan inhibited the generation of ${\cdot}$O2-,NO and ONOO- in the kidney of streptozotocin-induced diabetic rats. Renal aldose reductase and sorbitol dehydrogenase activities were increased in the streptozotocin-induced diabetic rats were reversed toward natural activities. Conclusions : Iksujisundan might inhibit the development of diabetes and its complications by scavenging reactive oxygen and nitrogen species, thereby by reducing oxidative stresses and also by regulating the activities of polyol pathway enzymes, all of which could help to recover kidney function.

• 동의생리병리학회지
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• 제21권3호
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• pp.671-678
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• 2007

Diabetes is a disease in which the body does not produce or properly use insulin. Etiological studies of diabetes and its complications showed that oxidative stress might play a major role. Therefore, many efforts have been tried to regulate free oxygen radicals for treating diabetes and its complications. Gamigukihwandong-hwan has been known to be effective for the treatment of diabetes. The present study was carried out to investigate the effect of Gamigukihwandong-hwan on renal function, peroxynitrite (ONOO$^-$) scavenging activity and polyol pathway in streptozotocin-induced diabetic rats. The crushed Gamigukihwandong-hwan was extracted 3 times, each time with 3 volumes of methyl alcohol at 60$^{\circ}C$ for 24 h. The extract was filtered and evaporated under a reduced pressure using a rotary evaporator to yield 74.95 g. Gamigukihwandong-hwan extract was oral-administered 100 mg per 1 kg of body weight for 20 days to the diabetic rats induced by streptozotocin (60 mg/kg). The effects of Gamigukihwandong-hwan extract on the streptozotocin-induced diabetic rats were observed by measuring the serum level of glucose, insulin, lipid components, creatinine and BUN, and also the kidney levels of superoxide anion radical (${\cdot}O_2^-$), nitric oxide (NO) and ONOO$^-$, and also the enzyme activities involved in polyol pathway. The Effects of Gamigukihwandong-hwan on the streptozotocin-induced diabetic rats with regards to body weight, blood glucose and insulin levels, creatinine and BUN levels, total cholesterol and triglyceride levels, and HDL-cholesterol levels were all shown to be good enough to cure and prevent the diabetes and its complications. Gamigukihwandong-hwan inhibited the generation of ${\cdot}O_2^-$, NO and ONOO$^-$ in the kidney of streptozotocin-induced diabetic rats. Renal aldose reductase and sorbitol dehydrogenase activities were increased in the streptozotocin-induced diabetic rats, whereas the ones in the Gamigukihwandong-hwan administered group among the streptozotocin-induced diabetic rats were reversed toward the natural activities. Gamigukihwandong-hwan might inhibit the development of diabetic nephropathy by scavenging reactive oxygen and nitrogen species, thereby by reducing oxidative stresses and also by regulating the activities of polyol pathway enzymes, all of which could help to recover the function of kidney.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2015년도 제49회 하계 정기학술대회 초록집
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• pp.159-159
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• 2015

A new approach for antimicrobial is based on the overproduction of reactive nitrogen species (RNS), especially; nitric oxide (NO) and peroxinitrite (ONOO-) are important factors to deactivate the bacteria. Recently, non-thermal atmospheric pressure plasma jet (APPJ) has been frequently used in the field of microbial sterilization through the generation of different kinds of RNS/ROS species. However, in previous study we showed APPJ has combine effects ROS/RNS on bacterial sterilization. It is not still clear whether this bacterial killing effect has been done through ROS or RNS. We need to further investigate separate effect of ROS and RNS on bacterial sterilization. Hence, in this work, we have enhanced NO production, especially; by applying a 1% of HNO3 vapour to the N2 based APPJ. In comparison with nitrogen plasma with inclusion of water vapour plasma, it has been shown that nitrogen plasma with inclusion of 1% of HNO3 vapour has higher efficiency in killing the E. coli through the high production of NO. We also investigate the enhancement of NO species both in atmosphere by emission spectrum and inside the solution by ultraviolet absorption spectroscopy. Moreover, qPCR analysis of oxidative stress mRNA shows higher gene expression. It is noted that 1% of HNO3 vapour plasma generates high amount of NO for killing bacteria.

• 대한한방내과학회지
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• 제32권1호
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• pp.56-67
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• 2011

Objectives : Recent etiological studies show that oxidative stress might play a major role in the diabetes and its complications. Mori Ramulus (MR) has been known to have antioxidative, anti-inflammatory and antidiabetic effects. The methanol extract of MR was tested for its effectiveness in LLC-PK1 cells exposed to high glucose. Methods : The cytoprotective effect of MR was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The antioxidative effect was measured in terms of generation amount of ${\cdot}O_2^-$ by 2',7'-dichlorodihydrofluorescein diacetate (DCFDA), NO by 4,5-diaminofluorescein (DAF-2), $ONOO^-$ by dihydrorhodamine 123 (DHR 123) in the high glucose -treated LLC-$PK_1$ cells. Western blotting was performed using anti-AGE, anti-RAGE, anti-MAPKs(ERK1/2, JNK, p38), anti-PI3K, anti-Akt, and anti-NF-${\kappa}$B (p50, p65) respectively. Results : MR extract reduced cell death and inhibited the generation of ${\cdot}O_2^-$, NO, $ONOO^-$ in the high glucose-treated LLC-$PK_1$ cells. MR inhibited the expression of AGE, RAGE, MAPKs, PI3K, and Akt by means of decreasing NF-${\kappa}$B activation. MR also inhibited NF-${\kappa}$B activation itself. Conclusions : These results indicate MR has cytoprotective, antioxidative, and anti-inflammatory effects. Therefore it is suggested that MR might prevent and cure diabetes and its complications.

• Preventive Nutrition and Food Science
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• 제18권1호
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• pp.18-22
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• 2013

To investigate radical scavenging effects and protective activities of bitter melon (Momordica charantia) against oxidative stress, in vitro and a cellular system using LLC-$PK_1$ renal epithelial cells were used in this study. The butanol (BuOH) fraction of bitter melon scavenged 63.4% and 87.1% of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals at concentrations of 250 and $500{\mu}g/mL$, respectively. In addition, the BuOH fraction of bitter melon effectively scavenged hydroxyl radicals (${\cdot}OH$). At all concentrations tested, the scavenging activity of the BuOH fraction was more potent than that of the positive control, ascorbic acid. Furthermore, under the LLC-$PK_1$ cellular model, the cells showed a decline in viability and an increase in lipid peroxidation through oxidative stress induced by pyrogallol, a generator of superoxide anion ($O_2{^-}$). However, the BuOH fraction of bitter melon significantly and dose-dependently inhibited cytotoxicity. In addition, 3-morpholinosydnonimine (SIN-1), a generator of peroxynitrite ($ONOO^-$) formed by simultaneous releases of nitric oxide and $O_2{^-}$, caused cytotoxicity in the LLC-$PK_1$ cells while the BuOH fraction of bitter melon ameliorated oxidative damage induced by $ONOO^-$. These results indicate that BuOH fraction of bitter melon has protective activities against oxidative damage induced by free radicals.