• 한국식품과학회지
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• 제46권3호
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• pp.375-383
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• 2014

본 연구는 복분자 완숙과 50% 에탄올 추출물을 이용하여 인간제대정맥내피세포주(HUVEC)에서 복분자 추출물이 NO의 생성에 미치는 영향과 ACE 단백질 발현을 조사하였으며, 본태성 고혈압 흰쥐(SHR)에서 혈압강하 효과 및 혈압의 조절과 관련 깊은 인자들(renin, AT1-R, ET-1, IL-6)의 유전자 발현 그리고 혈관조직의 변화를 조사하였다. 그 결과 복분자 완숙과 50% 에탄올 추출물은 HUVEC 세포주에서 NO의 생성을 촉진하였으며 혈관수축활성을 가지는 ACE 단백질의 발현을 억제하였고 본태성 고혈압 흰쥐에게 복분자 완숙과 에탄올 50% 추출물을 8주간 투여한 결과 혈액 내 ACE의 활성이 억제되었으며 renin 및 혈관 수축인자인 ET-1 유전자의 발현이 억제되고 염증관련 유전자인 IL-6의 발현이 억제됨을 확인하였다. 그리고 신세동맥의 두께 및 심근 호산성세포수를 감소시켜 혈관내피세포의 손상을 보호하며 심장의 병변을 경감시키는 효과가 있음을 확인하였다. 본 연구 결과 복분자 완숙과 추출물은 혈압 조절과 연관된 단백질 및 유전자의 발현과 활성을 조절하고, 혈관의 두께 및 심근의 변성을 감소시켜 혈압 강하에 도움이 되는 것으로 나타났으며 향후 심혈관계 질환을 개선시킬 수 있는 식품소재로서의 가능성이 있음을 확인하였다.

• 한국식품영양과학회지
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• 제44권1호
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• pp.7-13
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• 2015

본 연구에서는 아로니아 열매 추출물(AF-H)의 항염증 활성을 조사하기 위하여 LPS 자극에 의해 유도된 RAW 264.7 macrophage의 염증반응에서 AF-H의 염증매개인자 및 염증성 사이토카인 분비 억제활성과 이에 관련된 세포 내 작용기전 해석을 수행하였다. LPS($1{\mu}g/mL$)로 RAW 264.7 세포를 24시간 자극하는 염증모델에서 세포독성을 나타내지 않는 안전한 농도의 AF-H($0{\sim}500{\mu}g/mL$)를 LPS 처리 12시간 전에 처리하여 NO 및 PGE2의 분비 억제활성을 측정하였다. 그 결과 AF-H 처리에 의해 NO와 PGE2의 생성이 처리 농도에 의존하여 유의하게 억제되었으며, 이들 염증매개인자의 생합성 효소인 iNOS 및 COX-2의 세포 내 발현도 현저하게 억제되는 것으로 관찰되었다. 또한 AF-H의 처리에 의해 염증성 사이토카인인 $TNF-{\alpha}$와 IL-6의 분비도 유의하게 억제되는 것으로 확인하였다. 이러한 AF-H에 의한 항염증 활성의 세포 내 기전을 해석하기 위하여 LPS 자극에 의해 유도되는 MAPK와 $NF-{\kappa}B$ 전사인자의 활성화에 대한 억제 효과를 조사하였다. 그 결과 AF-H는 MAPK의 인산화에는 별다른 영향을 미치지 않고 $NF-{\kappa}B$의 활성화($I{\kappa}B$ 인산화)를 효과적으로 억제하는 것으로 확인되었다. 한편 LPS에 의한 in vivo 패혈증 모델에서 AF-H에 의한 패혈증 억제활성을 측정한 결과 비록 통계학적으로 유의하지는 않으나 AF-H 투여에 의해 생존율과 50% 사망률의 연장 효과가 관찰되었다. 이들 결과를 종합해 보면 아로니아 열매 열수추출물은 $NF-{\kappa}B$의 활성화 억제를 통해 NO, PGE2, $TNF-{\alpha}$ 및 IL-6 등의 염증매개인자와 사이토카인의 생성을 억제하는 항염증 활성을 지니는 것으로 확인되었다.

• 대한한의학방제학회지
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• 제26권4호
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• pp.295-306
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• 2018

Schisandra chinensis (SC) and Lycium chinense (LC) were widely distributed in Asia and the fruit has been used traditionally for medicinal herbs. The processing method was solid-state fermentation using Aspergillus oryzae for 48 h after stir-frying treatment at $220^{\circ}C$ for 12 min. In this study, in vitro the anti-inflammatory effect and in vivo hangover reduction were compared to unprocessed SC and LC water extract. Anti-inflammatory effects have been evaluated in pro-inflammatory mediators which were secreted by lipopolysaccharide (LPS)-induced RAW 264.7 macrophages. Nitric oxide (NO) was determined using Griess reaction. Proinflammatory cytokines such as tumor necrosis factor $(TNF)-{\alpha}$ and interleukin $(IL)-1{\beta}$ were measured by enzyme-linked immunosorbent assays (ELISA). Alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) activities were compared to processed SC or LC and mixtures thereof (1:1). In vivo study was compared to hangover relief in alcohol-fed mice. After administering a mixture of SC and LC (300 mg/kg) water extract (1:1), mice were fed 3 g/kg of ethanol. Serum was collected at 1, 3, and 5 h intervals to analyze ethanol and acetaldehyde levels using a colorimetric assay kit. The processed SC and LC water extracts compared to raw materials significantly inhibited LPS-induced NO and inflammatory cytokine production in RAW 264.7 cells. The results of the hangover mouse model are also consistent with anti-inflammatory effects. These results suggest that processed SC and LC extracts may be functional materials for the treatment of inflammation and hangover.

• 치위생과학회지
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• 제19권2호
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• pp.133-140
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• 2019

Background: The light-emitting diode (LED) curing light used is presumed to be safe. However, the scientific basis for this is unclear, and the safety of LED curing light is still controversial. The purpose of this study was to investigate the effect of LED curing light irradiation according to the conditions applied for the polymerization of composite resins in dental clinic on the cell viability and inflammatory response in Raw264.7 macrophages and to confirm the stability of LED curing light. Methods: Cell viability and cell morphology of Raw264.7 macrophages treated with 100 ng/ml of lipopolysaccharide (LPS) or/and LED curing light with a wavelength of 440~490 nm for 20 seconds were confirmed by methylthiazolydiphenyl-tetrazolium bromide assay and microscopic observation. The production of nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) was confirmed by NO assay and $PGE_2$ enzyme-linked immunosorbent assay kit. Expression of interleukin $(IL)-1{\beta}$ and tumor necrosis factor $(TNF)-{\alpha}$ in total RNA and protein was confirmed by reverse transcription polymerase chain reaction and Western blot analysis. Results: The LED curing light did not affect the viability and morphology of normal Raw264.7 cells but affected the cell viability and induced cytotoxicity in the inflammation-induced Raw264.7 cells by LPS. The irradiation of the LED curing light did not progress to the inflammatory state in the inflammation-induced Raw264.7 macrophage. However, LED curing light irradiation in normal Raw264.7 cells induced an increase in NO and $PGE_2$ production and mRNA and protein expression of $(IL)-1{\beta}$ and $(TNF)-{\alpha}$, indicating that it is possible to induce the inflammatory state. Conclusion: The irradiation of LED curing light in RAW264.7 macrophage may induce an excessive inflammatory reaction and damage oral tissues. Therefore, it is necessary to limit the long-term irradiation which is inappropriate when applying LED curing light in a dental clinic.

• Korean Journal of Acupuncture
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• 제36권1호
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• pp.63-73
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• 2019

Objectives : This study was designed to investigate the effects of Alismatis Rhizoma (AR) pharmacopuncture at BL23 on acute renal failure induced by cisplatin. Methods : The Sprague-Dawley rats were divided into Normal group (no injection of cisplatin and no treatment), Control group (cisplatin injection without treatment), Acu group (needling at BL23 after cisplatin injection), AR-PA1 group (treated with $0.3571mg/kg/20{\mu}l$ of AR pharmacopuncture at BL23 after cisplatin injection), and AR-PA2 group (treated with $1.7855mg/kg/20{\mu}l$ of AR pharmacopuncture at BL23 after cisplatin injection). Each treatment was given once daily for 8 days. Changes in body weight, kidney weight, tumor necrosis factor-alpha ($TNF-{\alpha}$), interleukin-6 (IL-6), Cu-Zn superoxide dismutase (Cu-Zn SOD), glutathione peroxidase (GPx), serum blood urea nitrogen (serum BUN), and serum creatinine were observed. Results : Body weight was significantly increased in AR-PA1 on $4^{th}$ and $6^{th}$ days and AR-PA2 on $2^{nd}$ day. $TNF-{\alpha}$ was significantly decreased in Acu, AR-PA1 and AR-PA2 groups. Cu-Zn SOD was significantly increased in AR-PA2 group. GPx was significantly increased in AR-PA1 and AR-PA2 groups. But kidney weight, IL-6, serum BUN and serum creatinine were not significantly changed in any groups compared to control group. Conclusions : In acute renal failure induced by cisplatin, AR pharmacopuncture has a mitigating effect on the inflammatory reaction related to the increase of $TNF-{\alpha}$ in the kidney tissue and a protective effect on the oxidative stress of the kidney tissue. However it is unlikely to restore the glomerular function or inhibit the renal swelling.

• 한국약용작물학회지
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• 제27권4호
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• pp.247-258
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• 2019

Background: Astragali radix (A) and Lithospermi radix (L) have long been used as traditional medicines due to their known anti-inflammatory effects. This study aimed at evaluating, their optimal mixing ratio and their functional compounds by investigating the inhibitory effects of mixed extracts of A and L and their active compounds on matrix metalloproteinases (MMPs). Methods and Results: A and L extracts were obtained by extraction at $80^{\circ}C$ using 50% and 70% fermented alcohol, respectively, and then mixed at a ratio of 5 : 5, 6 : 4, 7 : 3 and 8 : 2 (w/w). The activities of MMP-1, MMP-3, and MMP-13 were evaluated in interleukin-1beta ($IL-1{\beta}$)-induced SW1353 cells. The extract mixtures showed synergistic inhibitory effects on MMP-3 and MMP-13, higher than the effects of the individual A and L extracts. The 7 : 3 mixture (ALM16) showed the most effective MMPs inhibitory activity, while among the active ingredients, calycosin-7-O-${\beta}$-D-glucoside and lithospermic acid exhibited excellent MMPs inhibitory activity. Additionally, an HPLC method was established for simultaneous quantification of the effective components of the extract mixtures, and validated by measuring the linearity, precision and accuracy of the limit of detection (LOD) and limit of quantification (LOQ). Conclusions: ALM16 showed the most effective MMPs inhibitory activity. Calycosin-O-${\beta}$-D-glucoside, calycosin and lithospermic acid were identified as useful candidates, as they were the major functional compounds in the MMP inhibitory activity. Summarily, ALM16 might be a highly effective in osteoarthritis management, owing to its because it exhibits a protective effect on cartilage via excellent inhibition of MMPs.

• 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.

• 대한한방부인과학회지
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• 제32권2호
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• pp.1-17
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• 2019

Objectives: This study was performed to identify the anti-inflammatory effects of Salvia miltiorrhizae radix Water extract (SMW) on lipopolysaccharide (LPS) induced inflammation. Methods: RAW 264.7 cells were treated with 500 ng/ml of LPS. SMW (0.1, 0.25, 0.5 mg/ml) was treated 1 h prior to LPS. Cell viability was measured by MTT assay. Levels of nitric oxide (NO) were measured with Griess reagent and pro-inflammatory cytokines were measured by enzyme-linked immunosorbent assay (ELISA) and real-time polymerase chain reaction (PCR). We also examined molecular mechanisms such as mitogen-activated protein kinases (MAPKs) and nuclear factor-kappa B ($NF-{\kappa}B$) activation by western blot. In addition, we observed mice survival rate after LPS and examined their cytokine levels of serum and liver tissue. Results: SMW itself did not have cytotoxic effects in RAW 264.7 cells less than 0.5 mg/ml. SMW treatment inhibited the production of NO, and interleukin $(IL)-1{\beta}$ which is pro-inflammatory cytokine. And SMW treatment inhibited the LPS-induced activation of MAPKs such as extracellular signal-regulated kinase1/2 (ERK1/2), p38 kinases (p38), c-Jun NH2-terminal kinase (JNK) and $NF-{\kappa}B$. In addition, it also showed reducing the level of $IL-1{\beta}$ on the serum and liver tissue of mice. Also, death of LPS-induced mice was inhibited by SMW. Conclusions: The result suggests that treatment of SMW could reduce the LPS-induced inflammation. Thereby, SMW could be used as a protective agent against inflammation. Also, this study could give a clinical basis that SMW could be a drug or agent to prevent inflammatory diseases.

• Biomolecules & Therapeutics
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• 제29권6호
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• pp.685-696
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• 2021

In this study, we investigated the inhibitory effect of 5-aminolevulinic acid (ALA), a heme precursor, on inflammatory and oxidative stress activated by lipopolysaccharide (LPS) in RAW 264.7 macrophages by estimating nitric oxide (NO), prostaglandin E2 (PGE2), cytokines, and reactive oxygen species (ROS). We also evaluated the molecular mechanisms through analysis of the expression of their regulatory genes, and further evaluated the anti-inflammatory and antioxidant efficacy of ALA against LPS in the zebrafish model. Our results indicated that ALA treatment significantly attenuated the LPS-induced release of pro-inflammatory mediators including NO and PGE2, which was associated with decreased inducible NO synthase and cyclooxygenase-2 expression. ALA also inhibited the LPS-induced expression of pro-inflammatory cytokines, such as tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6, reducing their extracellular secretion. Additionally, ALA abolished ROS generation, improved the mitochondrial mass, and enhanced the expression of heme oxygenase-1 (HO-1) and the activation of nuclear translocation of nuclear factor-E2-related factor 2 (Nrf2) in LPS-stimulated RAW 264.7 macrophages. However, zinc protoporphyrin, a specific inhibitor of HO-1, reversed the ALA-mediated inhibition of pro-inflammatory cytokines production and activation of mitochondrial function in LPS-treated RAW 264.7 macrophages. Furthermore, ALA significantly abolished the expression of LPS-induced pro-inflammatory mediators and cytokines, and showed strong protective effects against NO and ROS production in zebrafish larvae. In conclusion, our findings suggest that ALA exerts LPS-induced anti-inflammatory and antioxidant effects by upregulating the Nrf2/HO-1 signaling pathway, and that ALA can be a potential functional agent to prevent inflammatory and oxidative damage.

• 치위생과학회지
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• 제20권4호
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• pp.221-229
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• 2020

Background: The purpose of this study was to investigate the anti-oral microbial activity and anti-inflammatory effects of rosmarinic acid (RA) in lipopolysaccharide (LPS)-stimulated MC3T3-E1 osteoblastic cells on a titanium (Ti) surface during osseointegration, and to confirm the possibility of using RA as a safe natural substance for the control of peri-implantitis (PI) in Ti-based dental implants. Methods: A disk diffusion test was conducted to confirm the antimicrobial activity of RA against oral microorganisms. In order to confirm the anti-inflammatory effects of RA, inflammatory conditions were induced with 100 ng/ml of LPS in MC3T3-E1 osteoblastic cells on the Ti surface treated with or without 14 ㎍/ml of RA. The production of nitric oxide (NO) and prostaglandin E2 (PGE2) in LPS-stimulated MC3T3-E1 osteoblastic cells on the Ti surface was confirmed using an NO assay kit and PGE2 enzyme-linked immunosorbent assay kit. Reverse transcription polymerase chain reaction and western blot analysis were performed to confirm the expression of interleukin (IL)-1β and tumor necrosis factor (TNF)-α in total RNA and protein. Results: RA showed weak antimicrobial effects against Streptococcus mutans and Escherichia coli, but no antimicrobial activity against the bacteria Aggregatibacter actinomycetemcomitans and the fungus Candida albicans. RA reduced the production of pro-inflammatory mediators, NO and PGE2, and proinflammatory cytokines, TNF-α and IL-1β, in LPS-stimulated MC3T3-E1 osteoblastic cells on the Ti surface at the protein and mRNA levels. Conclusion: RA not only has anti-oral microbial activity, but also anti-inflammatory effects in LPS-stimulated MC3T3-E1 osteoblasts on the Ti surface, therefore, it can be used as a safe functional substance derived from plants for the prevention and control of PI for successful Ti-based implants.