• Nutrition Research and Practice
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• 제12권6호
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• pp.486-493
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• 2018

BACKGROUND/OBJECTIVE: The honeysuckle berry (HB) contains ascorbic acid and phenolic components, especially anthocyanins, flavonoids, and low-molecular-weight phenolic acids. In order to examine the potential of HB as a hepatoprotective medicinal food, we evaluated the in vitro anti-oxidant and anti-inflammatory activities of Korean HB (HBK) and Chinese HB (HBC). MATERIALS/METHODS: Antioxidant and anti-inflammatory effects of the extracts were examined in HepG2 and RAW 264.7 cells, respectively. The anti-oxidant capacity was determined by DPPH, SOD, CAT, and ARE luciferase activities. The production of nitric oxide (NO) as an inflammatory marker was also evaluated. The Nrf2-mediated mRNA levels of heme oxygenase-1 (HO-1), NAD(P)H dehydrogenase [quinone] 1 (Nqo1), and glutamate-cysteine ligase catalytic subunit (Gclc) were measured. The concentrations of HB extracts used were 3, 10, 30, 100, and $300{\mu}g/mL$. RESULTS: The radical scavenging activity of all HB extracts increased in a concentration-dependent manner (P < 0.01 or P < 0.05). SOD (P < 0.05) and CAT (P < 0.01) activities were increased by treatment with $300{\mu}g/mL$ of each HB extract, when compared to those in the control. NO production was observed in cells pretreated with 100 or $300{\mu}g/mL$ of HBC and HBK (P < 0.01). Treatment with $300{\mu}g/mL$ of HBC significantly increased Nqo1 (P < 0.01) and Gclc (P < 0.05) mRNA levels compared to those in the control. Treatment with $300{\mu}g/mL$ of HBK (P < 0.05) and HBC (P < 0.01) also significantly increased the HO-1 mRNA level compared to that in the control. CONCLUSIONS: Thus, the Korean and Chinese HBs were found to possess favorable in vitro anti-oxidant and anti-inflammatory activities. Nrf2 and its related anti-oxidant genes were associated with both anti-oxidant and anti-inflammatory activities in HB-treated cells. Further studies are needed to confirm these in vivo effects.

• 생명과학회지
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• 제29권4호
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• pp.436-441
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• 2019

연은 아시아 국가에서 음식과 약재로 널리 사용되는 재료이다. 본 연구에서는 연의 잎(leaf, NL), 자육(seed, NS), 자방(seedpod, NSP)으로부터 에탄올 추출물을 제조하고 이들의 항염증 활성과 작용기전을 규명하였다. 이들의 항염증 활성을 연구하기 위하여 LPS로 자극된 RAW 264.7 세포에서 nitric oxide (NO) 생산을 측정하였다. NL, NS, NSP는 세포 생존율에 영향을 주지 않으면서, 농도의존적으로 NO의 생산을 현저하게 저해하였으며, iNOS 및 COX-2와 같은 pro-inflammatory 중재자들의 단백질 발현을 감소시켰다. 또한, NL, NS, NSP는 MAPKs 단백질의 인신화를 감소시키고 $NF-{\kappa}B$ p65의 핵으로의 이동을 저해함으로써, 세 추출물에 의한 항염증 활성은 MAPKs 경로와 $NF-{\kappa}B$ 경로를 조절함으로써 이루어짐을 제시한다. 게다가, ROS의 생성이 세 추출물에 의해서 모두 저해되었으며, HO-1의 발현과 HO-1의 전사조절인자인 Nrf2의 핵으로의 이동이 증가되었다. 결론적으로, 이러한 연구 결과는 연의 다양한 부위의 추출물인 NL, NS 그리고 NSP는 항염증 활성을 가지고 있으며, MAPKs, $NF-{\kappa}B$, Nrf2/HO-1 등 다양한 신호경로를 통해 조절할 수 있음을 제시한다.

• 한국식품과학회지
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• 제51권1호
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• pp.70-80
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• 2019

본 연구에서는 홍삼 비사포닌 분획(NSF)의 항 염증 효과를 마우스 대식세포와 인간유래 단핵세포에서 확인하였다. NSF는 마우스 대식세포에서 LPS로 유도된 NO, iNOS 그리고 COX-2의 양 뿐만 아니라 IL-6, $TNF-{\alpha}$, MCP-1과 같은 염증성 싸이토카인의 생성량을 유의적으로 감소시켰다. 인간 유래 단핵세포에서는 PMA에 의해 유도되는 대식세포로의 분화를 효과적으로 억제하면서 분화인자인 $CD11{\beta}$와 CD36의 발현을 유의적으로 감소시켰다. 마우스 대식세포에서와 마찬가지로 염증성 싸이토카인들의 생성량 또한 감소하였는데, 이러한 NSF의 항 염증 효과는 두 전사인자의 조절작용에 의한 것으로 사료된다. 즉 NSF는 $NF-{\kappa}B$의 핵으로 이동을 감소시킴으로써 전사활성을 억제하여 염증성 싸이토카인들의 발현을 저해하고 이와 반대로 Nrf2의 발현과 핵으로의 이동을 증가시켜 항산화 효소이면서 항 염증 작용을 나타내는 HO-1의 발현을 촉진하는 것으로 관찰되었다. 따라서 NSF는 $NF-{\kappa}B$와 Nrf2의 두 가지 신호전달체계를 조절함으로써 항 염증 작용을 나타냈으며 이를 홍삼 NSF의 항 염증 기작으로 보고하는 바이다.

• 생명과학회지
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• 제29권9호
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• pp.977-985
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• 2019

쑥부쟁이는 국화과에 속하는 다년생 식물로서 다양한 질병의 예방 및 치료에 오랫동안 사용되어 왔다. 최근 연구에서 쑥부쟁이 잎 추출물이 항산화 및 항염증 효과가 있는 것으로 알려져 있지만, 정확한 효능 평가에 관한 연구는 여전히 미비한 실정이다. 본 연구에서는 RAW 264.7 대식세포에서 쑥부쟁이 잎 에탄올 추출물(EEAY)의 항산화 효능이 항염증 효능과 연관이 있는지의 여부를 조사하였다. 본 연구의 결과에 의하면, EEAY는 $H_2O_2$ 처리에 의한 RAW 264.7 세포의 세포 독성을 유의적으로 억제시켰으며, 이는 Nrf2 및 HO-1의 발현 증가와 관련이 있음을 보여 주었다. 또한 EEAY는 $H_2O_2$에 의한 apoptosis를 유의적으로 억제하였으며, 이는 caspase-3의 활성 억제에 따른 PARP의 분해 차단과 연관성이 있었다. 그리고 EEAY는 대표적인 항 염증성 사이토카인인 IL-10의 발현 및 생산을 증가시켰으며, 이는 전사 및 번역 수준에서의 TLR-4 및 Myd88 발현 증가와 관련이 있었다. 아울러 EEAY는 LPS에 의한 염증성 매개인자인 NO의 생성 증가를 현저히 억제하였으며, EEAY에 의한 NO 생성의 억제 효과는 HO-1 유도제인 hemin에 의해 더욱 증가되었다. 따라서 본 연구의 결과는 EEAY에 의한 산화적 및 염증성 스트레스에 대한 RAW 264.7 대식세포의 보호 효과에 최소한 Nrf2/HO-1 신호 경로의 활성화가 관여할 가능성을 보여주었다.

• 대한한의학방제학회지
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• 제27권1호
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• pp.17-29
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• 2019

Inflammatory and oxidative stimuli play a critical role not only in the process of transforming normal cells into cancer cells, but also in the proliferation process of cancer cells. Sipyukmiryukieum (SYMRKU), a traditional Korean herb-combined remedy, is composed of 16 kinds of herbal medicines, which were recorded for "Ongjeo" treatment in "Dongeuibogam". In this study, we investigated the inhibitory effect of SYMRKU against inflammatory and oxidative responses in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Our results showed that SYMRKU significantly inhibited LPS-induced secretion of pro-inflammatory mediators including nitric oxide (NO) and prostaglandin $E_2$ without showing any significant cytotoxicity. Consistent with these results, SYMRKU down-regulated LPS-induced expression of their regulatory enzymes such as inducible NO synthase and cyclooxygenase-2. SYMRKU also inhibited LPS-induced production and expression of pro-inflammatory cytokines such as tumor necrosis factor-${\alpha}$, interleukin (IL)-$1{\beta}$ and IL-6. In addition, SYMRKU significantly reduced the production of reactive oxygen species by LPS and showed a strong, which was associated with induction of nuclear factor erythroid 2-related factor 2 and heme oxygenase-1 expression. Although further studies are needed to fully understand the anti-inflammatory effects associated with the antioxidant capacity of SYMRKU, the findings of the current study suggest that SYMRKU may have potential benefits by inhibiting the onset and/or treatment of inflammatory and/or oxidative diseases.

• 생명과학회지
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• 제31권2호
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• pp.158-163
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• 2021

본 연구에서는 풋사과의 열수추출물(GAHW)과 에탄올추출물(GAE), 그리고 애사과 열수추출물(UAHW)을 제조하고 LPS로 활성화된 RAW264.7세포주를 이용하여 이들의 항염증 활성을 연구하였다. 모든 추출물이 세포생존율에는 영향을 미치지 않고 nitric oxide (NO) 생산을 저해하였으며, iNOS의 발현도 감소시켰다. 반면, UAHW만이 COX-2의 발현을 저해하였다. 또한 모든 추출물이 모든 MAPKs의 인산화를 감소시켰다. 모든 추출물이 ROS의 생산을 농도의존적으로 감소시켰으며, GAHW와 GAE에 의해 HO-1의 발현이 증가됨을 확인하였다. 또한, 사과 flavonoid인 phloretin과 phloridzin의 항염증 활성을 연구하였다. Phloretin은 농도의존적으로 NO의 생산을 저해하였으나, phloridzin은 NO 생산에 아무 영향이 없었다. 또한, phloretin은 iNOS와 COX-2 단백질의 발현을 감소시킨 반면, phloridzin은 두 단백질 발현에 영향을 주지 못하였다. 따라서, 이러한 연구결과는 사과 추출물은 MAPK 경로와 HO-1 경로를 조절함으로써 항염증 활성을 가지며, phloretin이 사과의 항염증 활성을 담당하는 파이토케미칼의 하나가 될 수 있음을 제시한다.

• Biomolecules & Therapeutics
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• 제29권2호
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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.

• 생약학회지
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• 제51권4호
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• pp.244-254
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• 2020

Atractylodes macrocephala is a perennial herb and is a member of the Compositae family. This plant is known to contain various bioactive constituents indicating anti-inflammatory, neuroprotective, anti-oxidant, immunological enhancement, and gastroprotective effects. In this investigation, we isolated four compounds with similar chemical structures from A. macrocephala, and evaluated their anti-inflammatory effects. Among the four compounds, compound 2(atractylenolide II) showed the second-best inhibitory effect on the lipopolysaccharide(LPS)-induced production of nitric oxide in RAW264.7 macrophages and BV2 microglial cells. Compound 2 also inhibited the LPS-induced the production of prostaglandin E2(PGE2), and the expression of inducible nitric oxide synthase(iNOS) and cyclooxygenase(COX)-2 proteins in both cells. In addition, compound 2 suppressed the production of pro-inflammatory cytokines including interleukin(IL)-1β, IL-6, and tumor necrosis factor(TNF)-α. These inhibitory effects were contributed by inactivation of nuclear factor kappa B(NF-κB) and mitogen-activated protein kinases(MAPKs) pathways by treatment with compound 2. This compound did not induce the expression of heme oxygenase(HO)-1 protein indicating that the anti-inflammatory effect of compound 2 was independent with HO-1 protein. Taken together, these results suggested that atractylenolide II can be a candidate material to treat inflammatory diseases.

• 한방재활의학과학회지
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• 제31권1호
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• pp.17-32
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• 2021

Objectives The present study was designed to find out the therapeutic effects and possible underlying mechanism of Buja-tang, a herbal complex formula on experimental monosodium iodoacetate (MIA)-induced osteoarthritis. Methods Osteoarthritis models were created via intra-joint injection of MIA (50 μL with 80 mg/mL) in rats. Rats were divided into five groups and each group consisted of seven. Normal group was not injected MIA and did a normal diet. Control group injected MIA and received distilled water. Indo injected MIA and oral administration of 5 mg/kg of indomethacin. BJTL injected MIA and oral administration of 100 mg/kg of Buja-tang. BJTH injected MIA and oral administration of 200 mg/kg of Buja-tang. We analyzed weight-bearing ability of hind paws, oxidative stress related factor, antioxidant protein, inflammatory protein, inflammatory messenger and cytokine in joint tissue. Pathological observation of knee cartilage tissue structures was also performed with hematoxylin & eosin and safranin-O chromosomes. Results Weight-bearing ability of hind paws showed a tendency to reduce pain. The incidence of nicotinamide adenine dinucleotide phosphate oxidase and p22phox in articular tissue was significantly reduced, and the incidence of nuclear factor-erythroid 2-related factor 2 and heme oxygenase-1 and superoxide dismutases was significantly increased. The incidence of phosphorylated inhibitor of κBα, nuclear factor-kappa B p65, inducible nitric oxide synthase, cyclooxygenase-2, tumor necrosis factor alpha, interleukin (IL)-6, and IL-1β decreased significantly. In pathological observation, cartilage tissue damaged by MIAs in biopsy has significantly recovered from Buja-tang administration. Conclusions Buja-tang has anti-inflammation, antioxidation and pain relief effects. So this is thought to inhibit the progress of osteoarthritis in rat caused by the MIA.

• Journal of Ginseng Research
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• 제45권2호
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• pp.344-353
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• 2021

Background: Korean Red ginseng extract (KRGE) has beneficial effects on the cardiovascular system by improving endothelial cell function. However, its pharmacological effect on endothelial cell senescence has not been clearly elucidated. Therefore, we examined the effect and molecular mechanism of KRGE on the senescence of human umbilical vein endothelial cells (HUVECs). Methods: HUVECs were grown in normal or KRGE-supplemented medium. Furthermore, they were transfected with heme oxygenase-1 (HO-1) gene or treated with its inhibitor, a NF-κB inhibitor, and a miR-155-5p mimic or inhibitor. Senescence-associated characteristics of endothelial cells were determined by biochemical and immunohistochemical analyses. Results: Treatment of HUVECs with KRGE resulted in delayed onset and progression of senescence-associated characteristics, such as increased lysosomal acidic β-galactosidase and decreased telomerase activity, angiogenic dysfunction, and abnormal cell morphology. KRGE preserved the levels of anti-senescent factors, such as eNOS-derived NO, MnSOD, and cyclins D and A: however, it decreased the levels of senescence-promoting factors, such as ROS, activated NF-κB, endothelial cell inflammation, and p21 expression. The beneficial effects of KRGE were due to the induction of HO-1 and the inhibition of NF-κB-dependent biogenesis of miR-155-5p that led to the downregulation of eNOS. Moreover, treatment with inhibitors of HO-1, NF-κB, and miR-155-5p abolished the anti-senescence effects of KRGE. Conclusion: KRGE delayed or prevented HUVEC senescence through a signaling cascade involving the induction of HO-1, the inhibition of NF-κB-dependent miR-155-5p biogenesis, and the maintenance of the eNOS/NO axis activity, suggesting that it may protect against vascular diseases associated with endothelial senescence.