• 한국버섯학회지
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• 제17권3호
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• pp.136-143
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• 2019

국내 자생하는 한입버섯의 생리활성 및 항염 활성을 분석하기 위하여 열수 및 70% 에탄올, 70% 메탄올 추출물에 대한 DPPH 라디컬 소거능, 아질산염 소거능, 총 폴리페놀 함량, NO 생성률 및 세포생존율을 분석하였다. 본 연구결과 한입버섯의 열수추출물과 70% 메탄올 추출물에 비하여 70% 에탄올추출물의 DPPH 라디컬 소거능(76.1%), 아질산염 소거능(29.5%), 총 폴리페놀 함량(9.17 mg GAE/g)이 가장 높았다. 또한 총 폴리페놀 함량과 DPPH 라디컬 소거능 및 아질산염 소거능과는 유의적인 상관관계를 나타냈다. NO 생성률은 70%에탄올 추출물이 37.8%로 가장 낮아 NO 저해효능이 우수하였으며, 세포생존율은 81.4%로 열수추출물이 가장 높아 열수추출에 의한 세포독성이 낮은 것으로 나타났다. 또한 베타글루칸 함량은 37.8%로 측정되었다. 이와 같은 연구결과는 우리 주위에서 쉽게 접할 수 있는 야생버섯 중의 하나인 한입 버섯이 건강기능성 식품 등의 개발 가능성이 큰 버섯임을 보여주고 있으며, 일반인들도 차의 형태로 우려 마실 경우 여러 질병들에 대한 예방학적 차원에서 효과를 볼 수 있으리라 기대된다.

• Journal of Ginseng Research
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• 제43권3호
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• pp.377-384
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• 2019

Background: Inflammation is widespread in the clinical pathology and closely associated to the progress of many diseases. Triterpenoid saponins as a key group of active ingredients in Panax notoginseng (Burk.) F.H. Chen were demonstrated to show antiinflammatory effects. However, the chemical structures of saponins in the leaves and stems of Panax notoginseng (PNLS) are still not fully clear. Herein, the isolation, purification and further evaluation of the antiinflammatory activity of dammarane-type triterpenoid saponins from PNLS were conducted. Methods: Silica gel and reversed-phase C8 column chromatography were used. Furthermore, preparative HPLC was used as a final purification technique to obtain minor saponins with high purities. MS, NMR experiments, and chemical methods were used in the structural identifications. The antiinflammatory activities of the isolated saponins were assessed by measuring the nitric oxide production in RAW 264.7 cells stimulated by lipopolysaccharides. Real-time reverse transcription polymerase chain reaction was used to measure the gene expressions of inflammation-related gene. Results: Eight new minor dammarane-type triterpene oligoglycosides, namely notoginsenosides LK1-LK8 (1-8) were obtained from PNLS, along with seven known ones. Among the isolated saponins, gypenoside IX significantly suppressed the nitric oxide production and inflammatory cytokines including tumor necrosis $factor-{\alpha}$, interleukin 10, interferon-inducible protein 10 and $interleukin-1{\beta}$. Conclusion: The eight saponins may enrich and expand the chemical library of saponins in Panax genus. Moreover, it is reported for the first time that gypenoside IX showed moderate antiinflammatory activity.

• 한국응용과학기술학회지
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• 제38권5호
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• pp.1302-1313
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• 2021

본 연구는 가미수미원(백하수오 10 g,; 당귀 6 g; 구기자 8 g; 화피 6 g; 선태 3 g; 홍화 2 g)에 방기(Sinomenium acutum R.) 4 g의 첨가 여부에 따라 각각 열수추출물은 GS±SC(DW), 80% 에탄올 추출물은 GS±SC(EtOH)로 표기 하였다. 각 추출물은 항산화를 측정하기 위해 다음과 같은 연구를 수행하였다. HPLC를 통한 방기 지표물질 함량측정, 중금속 검사 기준 측정, ABTS/DPPH 라디칼 소거능 측정, SOD 유사 활성능 측정, ROS 및 NO 생성량을 측정하였다. 방기 지표물질 중 sinomenine의 함량은 열수 추출물에서, decursin은 에탄올 추출물에서 더 높게 확인되었다. 중금속 측정 결과 기준치 적합으로 확인되었다. ABTS/DPPD 라디칼 소거능을 측정한 결과 GS+SC(EtOH)의 소거능이 가장 높게 나왔으며, 상대적으로 열수보다 에탄올 추출이 라디칼 소거율이 높게 확인되었다. 그러나 SOD의 경우 열수 추출에서 상대적으로 유사 활성능이 높게 확인되었다. ROS 생성의 경우 열수 추출물이 에탄올 추출에 비해 유의하게 감소시켰으며, NO의 생성은 에탄올 추출물이 상대적으로 더 유의하게 감소시키는 것을 확인하였다. 본 연구 결과에서 GS+SC(EtOH)이 라디컬 소거능 및 NO 감소를 높게 나타낸 것으로 확인되어 산화적스트레스 후, 생성된 활성산소의 제거에 좀 더 영향을 미치는 것으로 판단된다. 향 후 지속적인 효능 심화 연구를 통해 조성물을 활용한 화장품 및 기능성 원료의 소재로서의 가능성이 높을 것으로 사료된다.

• 한국식품영양학회지
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• 제33권6호
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• pp.692-701
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• 2020

Centella asiatica (CA) has been widely used as herbal plants. It is a valuable resources. The aim of the present study was to evaluate physiological activities of solvent extracts from CA cultivated in Chungju, Korea (Good tiger care). After preparing water (cold-water, CA-WE; hot-water, CA-HWE) and EtOH extracts (50% EtOH, CA-50E; 70% EtOH, CA-70E), their total polyphenol, flavonoid, and triterpenoid contents, and anti-oxidant activities, and anti-inflammatory activities were examined and compared. CA-70E showed the most potent anti-oxidant activity based on ABTS radical scavenging activity and reducing ability assays, while CA-50E and CA-70E showed the highest DPPH radical scavenging activity. Among major triterpenoid glycosides present in CA, madecassoside and asiaticoside contents were found to be the highest in CA-70E, and madecassic acid and asiatic acid were the highest in CA-50E. In LPS-stimulated RAW 264.7 cells, CA-70E showed the highest TNF-α inhibitory activity, although CA-50E and CA-70E similarly inhibited nitric oxide production. Ethanol extracts significantly inhibited IL-6 and IL-8 production more than water extracts using TNF-α/IFN-γ-stimulated HaCaT cells, indicating its better inhibitory against skin inflammation. Therefore, Chungju-CA EtOH extract, especially 70% EtOH extract, has high physiologically active ingredients and potent anti-oxidant and anti-inflammatory activities, suggesting its industrial application as a functional material.

• Journal of Ginseng Research
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• 제45권1호
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• pp.48-57
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• 2021

Background: Ginsenoside Rh2 is well known for many pharmacological activities, such as anticancer, antidiabetes, antiinflammatory, and antiobesity properties. Glycosyltransferases (GTs) are ubiquitous enzymes present in nature and are widely used for the synthesis of oligosaccharides, polysaccharides, glycoconjugates, and novel derivatives. We aimed to synthesize new ginsenosides from Rh2 using the recombinant GT enzyme and investigate its cytotoxicity with diverse cell lines. Methods: We have used a GT gene with 1,224-bp gene sequence cloned from Lactobacillus rhamnosus (LRGT) and then expressed in Escherichia coli BL21 (DE3). The recombinant GT protein was purified and demonstrated to transform Rh2 into two novel ginsenosides, and they were characterized by nuclear magnetic resonance (NMR) techniques and evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2-5-diphenyltetrazolium bromide assay. Results: Two novel ginsenosides with an additional glucopyranosyl (6→1) and two additional glucopyranosyl (6→1) linked with the C-3 position of the substrate Rh2 were synthesized, respectively. Cell viability assay in the lung cancer (A549) cell line showed that glucosyl ginsenoside Rh2 inhibited cell viability more potently than ginsenoside Rg3 and Rh2 at a concentration of 10 μM. Furthermore, glucosyl ginsenoside Rh2 did not exhibit any cytotoxic effect in murine macrophage cells (RAW264.7), mouse embryo fibroblasts cells (3T3-L1), and skin cells (B16BL6) at a concentration of 10 μM compared with ginsenoside Rh2 and Rg3. Conclusion: This is the first report on the synthesis of two novel ginsenosides, namely, glucosyl ginsenoside Rh2 and diglucosyl ginsenoside Rh2 from Rh2 by using recombinant GT isolated from L. rhamnosus. Moreover, diglucosyl ginsenoside Rh2 might be a new candidate for treatment of inflammation, obesity, and skin whiting, and especially for anticancer.

• 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.

• 한방재활의학과학회지
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• 제31권1호
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• pp.33-46
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• 2021

Objectives The aim of this study is to evaluate anti-inflammatory and anti-arthritic effects of Sogyunghwalhyel-tang-gamibang (SGHHTGB) in cell and animal models and also to suggest one of putative mechanisms underlying its anti-arthritic effects. Methods Enzyme-linked immunosorbent assay was applied to measure the concentrations of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6 and prostaglandin E2 (PGE2) in culture medium and blood serum and nitric oxide (NO) was assayed by Griess reagent. The expressions of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) were analyzed by Western blot method. Results In a cell model using RAW264.7 macrophages stimulated with the endotoxin lipopolysaccharide (LPS), the drug, at its non-cytotoxic concentrations, inhibited the production of the pro-inflammatory cytokine TNF-α, IL-1β and IL-6. In addition, it suppressed the expression of the inflammatory enzyme iNOS and COX-2, and reduced the synthesis of the enzyme product NO (as stable nitrite) and PGE2 in activated macrophages. Meanwhile, in an animal model using rheumatic arthritis (RA) mice induced with injection of type II collagen antibody (CAb) and LPS, the drug improved clinical symptom of arthritis and reduced paw thickness and inflammatory cell infiltration. In blood of RA mice, the drug reduced serum levels of TNF-α, IL-1β, IL-6, nitrite, and PGE2, all inflammatory mediators produced by activated macrophages. Conclusions SGHHTGB may ameliorate CAb and LPS-induced RA in mice, presumably by inactivating macrophages that are capable of initiating joint inflammation by producing pro-inflammatory cytokines and expressing inflammatory enzymes.

• 대한한의학방제학회지
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• 제29권2호
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• pp.81-91
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• 2021

Objectives : This study was conducted to evaluate the efficacy of a complex mixture of natural substances of ginseng and baeknyeoncho on the arthritic rats. Methods : In vitro experiments were conducted to ensure the stability of the complex. After setting toxicity and concentration by MTT assay, the antioxidant effect was measured through DPPH and ABTS radical scavenging activity. To confirm the anti-inflammatory effects of the complex, levels of nitric oxide (NO) and pro-inflammatory cytokines (IL-1β, TNF-α) were measured in LPS-treated macrophage cell lines (RAW264.7). We injected monosodium iodoacetate (MIA) 50 μl (60 mg/ml) into knee joints of rats to induce osteoarthritis. The rats were divided into three groups (normal (n=5), control (n=5), and OR (n=5) group). The control group consumed 2 mg/kg of physiological saline once a day for 4 weeks, and the OR group was mixed at a concentration of 416.5 mg/kg of Baengnyeoncho (O) and 208.25 mg/kg of red ginseng (R) and ingested 1 mL each 5 days a week. Results : This complex increased the DPPH and ABTS radical scavenging rate. The complex decreased NO production and pro-inflammatory cytokine production of macrophages. In the OR group, the secretion of cytokine in serum was decreased. In histopathological examination, the joint tissue of the composite showed less damage to the synovial membrane, cartilage, and fibrous tissue than the control group. Conclusions : As a result of this study, natural complexes have antioxidant, anti-inflammatory and cartilage protection effects. Therefore, we expect the complex to be effective in treating osteoarthritis.

• 한국식품과학회지
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• 제54권2호
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• pp.147-154
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• 2022

본 연구에서는 몽골 야생 베리 추출물의 총 페놀성 화합물, 플라보노이드, 그리고 안토시아닌 함량과 항산화 효과를 측정하였다. 전구 염증매개 사이토카인의 유전자 발현에 미치는 영향을 살펴본 결과, LBE와 BBE의 전처리가 LPS 자극에 의해 증가된 TNF-α, IL-1β, iNOS, 및 COX-2의 mRNA 발현의 농도를 유의적으로 감소시키며 항염증 효능을 보여주었다. 또한 LBE와 BBE는 NOX2의 발현 억제를 통하여 활성산소의 생성을 저해시킴으로 세포내 염증성 산화적 스트레스를 감소시킴을 확인하였다. 몽골야생 블루베리에서는 6종의 안토시아닌이 검출되었으며, 야생 링곤베리에서는 1종의 안토시아닌이 검출되었다. 본 연구결과를 토대로 몽골의 야생 베리가 강력한 항산화 및 항염증을 통한 육식위주의 식단을 소비하는 몽골인들의 산화적 스트레스 및 만성 염증성 질환을 줄여주는 식품으로의 가능성을 확인하였다.

• 현장농수산연구지
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• 제23권1호
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• pp.31-36
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• 2021

고 문헌에 기록된 부자의 가공수치 포제법으로 숙부자로 가공하고 인삼, 생강, 감초의 배합으로 폐암세포와 대식세포 독성에 관하여 실험한 결과 1번째 실험군 인삼과 숙부자 처리군에서 100 ㎍/ml 농도에서 독성에 영향을 주기 시작하여 1000 ㎍/ml 농도에서 더 많은 영향을 주었다. 그 중 대식세포 보다는 A549 폐암세포에서 더 많은 세포독성에 영향을 주었다. 그리고 2번째 실험군에 추가로 생강이 배합된 처리군에서는 1000 ㎍/ml 농도부터 세포독성이 시작 되었으며 대식세포 보다는 A549 폐암세포에 세포독성에 더 많은 영향을 주었으며 마지막 3번째 실험군 생강과 감초가 추가된 실험군에서는 100 ㎍/ml 농도에서 A549 폐암세포에 독성이 시작 되었지만 대식세포에는 영향이 없었으며, 1000 ㎍/ml 농도부터 대식세포와 A549 폐암세포에 영향을 주었다. 결론적으로 독성이 강한 부자를 사용함에 있어서는 반드시 숙부자로 가공하여 활용 하여야 하며 인삼, 생강, 감초의 배합으로 정상세포인 대식세포 독성에는 적은 영향과 특정 폐암세포가 더 많은 세포독성이 있음을 확인할 수 있었다. 앞으로도 인삼, 생강, 감초의 복합추출물이 지속적인 연구를 통해 항암활성의 효능을 검증하고 추가적인 연구가 필요할 것이라 사료된다.