• Nutrition Research and Practice
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• 제10권2호
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• pp.131-138
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• 2016

BACKGROUND/OBJECTIVES: Citrus and its peels have been used in Asian folk medicine due to abundant flavonoids and usage of citrus peels, which are byproducts from juice and/or jam processing, may be a good strategy. Therefore, the aim of this study was to examine antioxidant and anti-inflammatory effects of bioconversion of Jeju Hallabong tangor (Citrus kiyomi ${\times}$ ponkan; CKP) peels with cytolase (CKP-C) in RAW 264.7 cells. MATERIALS/METHODS: Glycosides of CKP were converted into aglycosides with cytolase treatment. RAW 264.7 cells were pre-treated with 0, 100, or $200{\mu}g/ml$ of citrus peel extracts for 4 h, followed by stimulation with $1{\mu}g/ml$ lipopolysaccharide (LPS) for 8 h. Cell viability, DPPH radical scavenging activity, nitric oxide (NO), and prostagladin $E_2$ ($PGE_2$) production were examined. Real time-PCR and western immunoblotting assay were performed for detection of mRNA and/or protein expression of pro-inflammatory mediators and cytokines, respectively. RESULTS: HPLC analysis showed that treatment of CKP with cytolase resulted in decreased flavanone rutinoside forms (narirutin and hesperidin) and increased flavanone aglycoside forms (naringenin and hesperetin). DPPH scavenging activities were observed in a dose-dependent manner for all of the citrus peel extracts and CKP-C was more potent than intact CKP. All of the citrus peel extracts decreased NO production by inducible nitric oxide synthase (iNOS) activity and $PGE_2$ production by COX-2. Higher dose of CKP and all CKP-C groups significantly decreased mRNA and protein expression of LPS-stimulated iNOS. Only $200{\mu}g/ml$ of CKP-C markedly decreased mRNA and protein expression of cyclooxygenase-2 in LPS-stimulated RAW 264.7 cells. Both 100 and $200{\mu}g/ml$ of CKP-C notably inhibited mRNA levels of $interleukin-1{\beta}$ ($IL-1{\beta}$) and IL-6, whereas $200{\mu}g/ml$ CKP-C significantly inhibited mRNA levels of $TNF-{\alpha}$. CONCLUSIONS: This result suggests that bioconversion of citrus peels with cytolase may enrich aglycoside flavanones of citrus peels and provide more potent functional food materials for prevention of chronic diseases attributable to oxidation and inflammation by increasing radical scavenging activity and suppressing pro-inflammatory mediators and cytokines.

• 대한한의학방제학회지
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• 제28권4호
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• pp.339-349
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• 2020

Objectives : Wiryeong-tang (WRT) is a traditional herbal medicine used to treat kidney-related diseases. However, the anti-inflammatory and anti-gastritis effect of Wiryeong-tang was not well known. Therefore, we experimented to confirmed the anti-inflammatory and anti-gastritis effects of Wiryeong-tang. Methods : The RAW 264.7 cells were pre treated with Wiryeong-tang mix soft extract (WRT-mse; 50, 100 and 200 ㎍/mL) for 1 hrs, and then incubated with lipopolysaccharide (LPS; 500 ng/mL). Cell viability was measured by the MTT method, and nitric oxide (NO) was measured with griess reagent. In addition, pro-inflammatory cytokines were measured by enzyme-linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR). For anti-gastritis effect in vivo, acute gastritis was induced using 150 mM HCl/60% ethanol used ICR mice. WRT-mse (133 mg/kg) was pre treated for 3 days and then treated with 150 mM HCl/60% ethanol 1 hrs later. Then gastritis was observed and inflammatory cytokines in the gastric tissue was measured. Results : The 8 marker components of the WRT-mse were determined by simultaneous analysis using HPLC. WRT-mse was not toxic and inhibited pro-inflammatory cytokines such as IL-1β, IL-6 and TNF-α at NO production, protein and mRNA levels. Also, it was confirmed that WRT-mse improved bleeding and edema in gastritis, and suppresses inflammatory cytokines. Conclusion : In summary, our results suggest that the treatment of the WRT-mse reduced and improved the 150 mM HCl/60% ethanol induced acute gastritis and the inflammation caused by LPS stimulation in RAW 264.7 cells. Therefore, this study may provide useful drug or clinical evidence for WRT-mse to prevent inflammation.

• Journal of Nutrition and Health
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• 제40권7호
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• pp.624-629
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• 2007

생체 내 (in vivo) 실험에서 톳 추출물을 2주 동안 격일로 0, 50, 500 mg/kg B.W.의 농도로 마우스에 경구투여한 후 비장세포 증식능 및 복강 대식세포에서 분비하는 cytokine ($IL-1{\beta}$, IL-6, $TNF-{\alpha})$ 생성량을 검색한 결과 50 mg/kg B.W.과 500 mg/kg B.W.의 농도에서 Con A나 LPS로 자극 시 대조군에 비해 높은 비장세포 증식능을 보였고 특히 50 mg/kg b.w. 농도로 투여한 군에서 비장세포 증식능이 최대를 나타내는 것을 알 수 있었다. 복강 대식세포에 의한 cytokine 생성량을 측정한 결과 LPS 첨가시 $IL-1{\beta}$의 분비량은 2주 경구 투여 시에 50 mg/kg B.W.의 농도군에서 가장 높은 생성량을 보였다. 이는 LPS를 투여한 군과 투여하지 않은 군에서 모두 동일한 경향을 보였다. IL-6 분비량은 LPS 첨가시 50 mg/kg B.W.의 농도 투여군에서 가장 높은 생성을 나타냈다. $TNF-{\alpha}$의 경우는 유의적인 차이를 보이지 않았다. 이상의 결과로 톳 열수 추출물은 대식세포의 활성화에 작용하여 사이토카인 생성을 증가시키는데 영향을 미칠 수 있는 것으로 예측된다. 따라서 톳 추출물은 마우스 면역계의 조절기전에 작용하여 비장세포와 대식세포의 활성화를 유도함으로써 면역세포 활성을 직접적으로 촉진시키거나 또는 이와 관련된 다른 면역반응에 영향을 미침으로서 면역 활성에 효과적으로 작용할 가능성이 있으리라 사료된다.

• KSBB Journal
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• 제23권2호
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• pp.164-168
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• 2008

본 연구를 통하여 요즘 기능성 측면에서 관심을 끌고 있는 참죽나무 잎의 열수추출물과 유기용매 추출물을 이용하여 항산화능과 면연증강능을 확인하였다. 열수 추출물의 총 폴리페놀 함량은 46.5-59.6 mg/100 g으로 유기용맹 추출물보다 그 수치가 더 높았다. 열수 추출물과 유기용맹 추출물의 항산화 활성은 2,2-diphenyl-picryl-hydraryl (DPPH) radical 소거활성으로 6-33%로 기준물질보다 낮은 값을 나타내었다. 참죽나무 잎 추출물의 면역증강능을 확인하기 위하여 박테리아 감염에 대항하는 숙주방어체계에 필요한 NO 생성능을 확인하였다. 열수 추출물은 마크로파지 세포주인 RWA 264.7의 NO 생성을 증가시켰다. 그러나 유기용매추출물은 별다른 변화를 가져오기 않았다. 항염효과를 증명하기 위하여 인지질다당류 (LPS) 유발 NO생성에 미치는 영향을 조사하여, NO 생성을 억제하는 것을 증명하였다. 즉, 면역증강능의 확인 실험을 통하여 참죽나무 잎의 열수추출 분획에 대식세포의 활성을 증가시키는 성분이 포함되어 있고, 유기용매 추출 분획에서는 대식세포의 활성을 감소시키는 성분이 포함되어 있는 것으로 판단된다. 이상과 같은 결과를 미루어 보았을 때, 참죽이 한 가지 식물인데도 불구하고 유효 성분에 대한 추출 방법에 따라 상이한 면역 증강 및 항균, 항염 작용을 보이는 것으로 나타났다.

• 대한한의학회지
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• 제30권4호
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• pp.13-27
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• 2009

Objectives: Trans-cinnamaldehyde (TCA) is the main component of Cinnamomi Ramulus and it has been reported that TCA inhibits inflammatory responses in various cell types. Inflammation-mediated neurological disorders induce the activation of macrophages such as microglia in brain, and these activated macrophages release various inflammation-related molecules, which can be neurotoxic if overproduced. In this study, we evaluated gene expression profiles using gene chip microarrays in lipopolysaccharide (LPS)-stimulated BV-2 cells to investigate the antiinflammatory effect of TCA on inflammatory responses in brain microglia. Methods: A negative control group was cultured in normal medium and a positive control group was stimulated with $1{\mu}g/ml$ in the absence of TCA. TCA group was pretreated with $10{\mu}g/ml$ before $1{\mu}g/ml$ LPS stimulation. The oligonucleotide microarray analysis was performed to obtain the expression profiles of 28,853 genes using gene chip mouse gene 1.0 ST array in this study. Results: In positive control group, 1522 probe sets were up-regulated in the condition of the cutoff value of 1.5-fold change and 341 genes with Unigene ID were retrieved. In TCA group, 590 probe sets were down-regulated from among 1522 probe sets and 33 genes with Unigene ID were retrieved, which included 6 inflammation-related genes. We found out that Id3 gene is associated with transforming growth factor-${\beta}$ (TGF-${\beta}$) signaling pathway and Klra8 gene is related to natural killer cell-mediated cytotoxicity pathway. Conclusions: The results mean that TCA inhibits inflammatory responses through down-regulating the expressions of inflammation-related genes in LPS-stimulated BV-2 cells.

• 동의생리병리학회지
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• 제24권2호
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• pp.310-318
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• 2010

Dioscorea bulbifera is one of the traditional medicinal herb. It commonly used in the treatment of hematemesis, epistaxis, tuberculous cervical lymphadenitis, laryngitis, acute infectious disease in East Asia. In the present study, we have demonstrated the anti-inflammatory effects of Dioscorea bulbifera MeOH extract (DBME) in macrophage cell line. To investigate mechanism of the anti-inflammatory activity, we examined the effects of the lipopolysaccaride (LPS)-induced production of nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$), pro-inflammatory cytokines and expression of inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), p-inhibitory ${\kappa}B{\alpha}$ (p-$I{\kappa}B{\alpha}$), and nuclear factor-${\kappa}B$ (NF-${\kappa}B$) in a murine macrophage cell line RAW 264.7. The RAW 264.7 cells were cultured in DMEM + serum medium for 24 hrs. After serum starvation for 24 hrs, the cells were treated with DBME 0.03, 0.10, 0.30 mg/$m{\ell}$ for 1 h, followed by stimulation with LPS (1 ${\mu}g/m{\ell}$) for activation of immune response. After treatment, cell viability was measured by MTT assay, and NO production was monitored by measuring the nitrite content in culture medium. The protein band of iNOS, COX-2, p-$I{\kappa}B{\alpha}$, and NF-${\kappa}B$ was determined by immunoblot analysis and levels of cytokine were analyzed by sandwich immunoassays. There were three experimental groups: carrageenan, DBME 0.3, 1.0 g/kg. Rats were administrated either carrageenan (40% PEG) or carrageenan + DBME (0.3, 1.0 g/kg body weight) for 4 days (p.o.). To induce acute paw edema, rats were injected 1% carrageenan (100 ${\mu}{\ell}$/rat, dissolved in sterilized saline). The effect of DBME in the carrageenan-induced rat paw edema. As results, DBME has an inhibitory effect on the production of NO, PGE2, TNF-${\alpha}$, IL-$1{\beta}$ and IL-6 and on the expression of iNOS, COX-2, p-$I{\kappa}B{\alpha}$ and translocation of NF-${\kappa}B$ to nuclear from cytosol. In addition, DBME effectively inhibited the increases of paw edema induced by carrageenan treatment in vivo. These results suggest that DBME can inhibit production of pro-inflammatory mediators and might be a useful source for treatment of acute inflammatory disease.

• 생명과학회지
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• 제27권10호
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• pp.1130-1136
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• 2017

본 연구에서는 장수풍뎅이 유충 추출물을 이용하여 알레르기 및 염증 반응에 미치는 효과를 확인하였다. 장수풍뎅이 유충 추출물의 항알레르기와 항염증 효능은 Compound 48/80에 의해 활성화된 비만세포(RBL-2H3)와 lipopolysaccharide(LPS)로 활성화된 대식세포(Raw 264.7)로부터 분비 또는 발현되는 ${\beta}-hexosaminidase$, $TNF-{\alpha}$, IL-4, IL-6, COX-2, nitric oxide, 및 iNOS를 측정하여 관찰하였다. 비만세포에 장수풍뎅이 유충 추출물과 Compound 48/80을 함께 처리한 경우 비만세포의 탈과립에 의해서 분비되는 ${\beta}-hexosaminidase$의 양이 현저히 감소하였으며 $TNF-{\alpha}$, IL-4, COX-2의 발현 또한 효과적으로 감소시킴을 확인할 수 있었다. 또한 LPS에 의해 활성화된 대식세포(Raw 264.7)는 장수풍뎅이 유충 추출물 처리에 의해 염증반응인자인 IL-6와 nitric oxide의 분비량이 현저히 감소함을 확인할 수 있었다. 이상의 결과는 장수풍뎅이 유충 추출물이 알레르기와 염증반응의 원인 물질인 ${\beta}-hexosaminidase$ 억제 효과뿐만 아니라, 염증 cytokine의 발현을 저해하는 것으로 보아 알레르기와 염증 질환의 예방과 치료에 효과적으로 이용될 수 있음을 확인할 수 있었다.

• 동의생리병리학회지
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• 제34권6호
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• pp.348-354
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• 2020

Oryeong-san (ORS) is a traditional Korean herbal medicine widely used for renal associated diseases, composed of five medicine herbs; Atractylodes japonica Koidzumi, Cinnamomum cassia Presl, Polyporus umbellatus Fries, Poria cocos Wolf and Alisma orientale Juzepzuk. We studied to improve the convenience of intake and portability by developing modernized dosage forms, and examined the effect on anti-inflammation of ORS. In order to develop the tablet formulation of ORS (ORS-F), the tablets were evaluated on the basis of physical characteristics include diameter, thickness, weight variation, hardness, friability and disintegration. To analyze the marker components of ORS-F, eight index markers from five herbal medicines were chosen. And the method using high performance liquid chromatography (HPLC) with diode-array detector method was established for the simultaneous analysis. The biological activities were examined the effect of ORS-F on pro-inflammation mediated by LPS-stimulation. The production of nitric oxide (NO) and cytokines were determined by reacting cultured medium with griess reagent and enzyme-linked immunosorbent assay (ELISA). The expression of cyclooxygenase-2 (COX-2) and inducible NO synthase (iNOS) were investigated by Western blot and RT-PCR. The anti-oxidant activities of OJS-F increased markedly, in a dose-dependent manner. and, The total phenolic compound and flavonoids contents of OJS-F were 10.20±0.09 ㎍/㎎ and 12.86±0.86 ㎍/㎎. OJS-F which is LPS has diminished in the LPS-induced release of inflammatory mediators (NO, iNOS, COX2 and PGE2) and pro-inflammatory cytokines (TNF-α, IL-6 and IL-1β) from the RAW264.7 macrophages. Therefore, the developed formulation for tablet of ORS-F provide efficiency and usability, and indicated effect of anti-inflammation.

• Tuberculosis and Respiratory Diseases
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• 제44권2호
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• pp.379-390
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• 1997

연구배경 : 급성호흡곤란증후군의 병인론은 확실치 않으나 일반적으로호중구와 호중구에서 생산된 매개물들이 중심적 역할을 하는 것으로 알려져 있다. 그러나 지속적이며 심한 호중구강소증에도 불구하고 실제 임상에서 급성 호흡증후군이 발생한 경우가 상당수 보고되어 있으며 특히 환자의 병리조직학적 검사상에서도 침윤이 없었다는 잠을 고려해볼때 급성호흡곤란증후군의 병리기전에 호중구외의 다른 효과세포가 작용하고 있음을 알 수 있다. 따라서 저자들은 백서에 약물을 투여하여 호중구를 거의 제거한다음 호중구 감소증하에서 내독소에 의한 급성폐손상의 발병기전을 알아 보고자하였다. 방 법 : 웅성 Sprague-Dawley률 정상 대조군과 (내독소군), cyclophosphamide (CPA)를 주입하여 호중구를 결핍시킨군 (CPA군)으로 분류하였다. LPS 5mg/kg를 미부정맥을 통하여 주입하여 급성폐손상를 유발시킨후 3시간 및 6시간째에 급성폐손상지표로서 기관지폐포세척액내 단백질 농도를 측정하였고 병리기전을 이해하기 위하여 기관지폐포세척액내 염증 세포의 변화를 관찰하고 TNF-alpha와 IL-6를 측정하였으며 기관지폐포세척액내 염증세포의 과산화수소 분비능을 각군간에서 비교하였다. 결 과 : Cyclophosphamide (CPA) 투여후 말초혈액내 백혈구수의 변화CPA투여후 전체 백혈구와 호중구수가 각각 95%이상 감소하였다. 기관지폐포세척액내의 전체및 감별 세포수의 변화내 독소군에서는 총세포수가 3시간 및 6시간째 모두 정상군에 비해 매우 유의하게 상승하였으나 (p < 0.01) CPA군에서는 세포종가를 관찰할수 없었다. 내독소군에서 대식세포수와 호중구수는 모두 증가를 보였으나 (p < 0.05), CPA군에서는 대조군과 비교시 대식세포의 수와 구성비는 차이가 없었고 호중구의 수가 매우 감소하여 (p < 0.01), 대식세포가 99.7%이상을 차지하였다. 기관지폐포세척액내의 단백질 농도의 변화 : 내독소군에서는 3시간째부터 유의한 증가를 보였고 (p < 0.05), 6시간째에는 더욱 증가하였다 (p < 0.01). CPA군 역시 3시간째에 단백질농도가 대조군에 비해 유의하게 상승되었으나 (p < 0.05) 내독소군과는 차이가 없었으며, 6시간째에는 내독소군에 비해 유의하게 감소하였으나 (p < 0.05), 대조군에비해서는 여전히 높은 값을 보였다 (p < 0.05). 기관지폐포세척액내의 cytokine농도의 변화 종양괴사인자와 IL-6는 내독소군 및 CPA군 모두에서 대조군에 비해 유의하게 증가되었고 내독소군과 CPA군간의 차이는 없었다. 기관지폐포세척액내 염증세포의 과산화수소 분비능 측정 각군에서 상태에서는 서로 유의한 차이가 없었으나 zymosan으로 자극한 상태에서는 모두 자극전에 비해 유의한 차이로 증가하였으며 내독소군의 경우 약 89.0%로서 가장 큰 증가를 보였고 (p < 0.0008), CPA군도 42.85로서 내독소군에 비해서는 그 정도가 다소 작았으냐 (p = 0.033) 대조군에 비해서는 역시 큰 증가를 보였다 (p = 0.003). 결 론 : 호중구 결핍상태의 급성폐손상은 단핵구, 그 중에서도 기존의 폐포대식세포의 활성화에 있는 것으로 생각된다.

• BMB Reports
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• 제48권5호
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• pp.239-240
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• 2015

Dysregulation of cytokine expression causes inflammatory diseases or chronic infection conditions. We have identified that Tat-activating regulatory DNA-binding protein-43 (TDP-43) is involved in cytokine RNA processing in order to promote an optimal immune response. The interaction of TDP-43 with spliceosomal components from the Cajal body leads to the formation of a novel sub-nuclear body called the Interleukin (IL)-6 and IL-10 Splicing Activating Compartment (InSAC). TDP-43 binds to the IL-6 and IL-10 RNAs in a sequence-dependent manner. In cell-based studies, we observed that lipopoly-saccharide (LPS) stimulation induces the formation of the InSAC through TDP-43 ubiquitination, thereby influencing the processing and expression levels of IL-6 RNA. Moreover, TDP-43 knockdown in vivo results in a decrease in IL-6 production and its RNA splicing and stability. Thus, these findings demonstrate that the InSAC is linked to the activation and modulation of the immune response. [BMB Reports 2015; 48(5): 239-240]