• Journal of Applied Biological Chemistry
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• 제65권2호
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• pp.113-120
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• 2022

구슬말(Nostoc commune Vaucher ex Bornet & Flahault)은 이형세포를 갖는 특징으로 다른 목들과 구분되는 남조류의 일종으로 극지방에서 열대지역까지 광범위한 지역에 분포하며 단세포가 연결되어 형성된 수많은 trichome들이 점액질에 둘러 쌓인 형태로 커다란 군체를 형성한다. 주로 토양, 암반, 잔디 위 등에 서식한다고 알려져 있으나 흔히 관찰되지 않기 때문에 현재 연구가 거의 없는 실정이다. 이에 본 연구에서는 토양 남조류인 N. commune HCW0811을 분리 및 동정하였으며 항염증 활성을 조사 하고자 하였다. 그 결과 N. commune HCW0811는 LPS로 유도된 RAW 264.7세포에서 80%이상의 세포 생존율을 나타내었으며 NO, PGE2 및 TNF-α, IL-6, IL-1β의 생성을 효과적으로 억제하였다. 또한 western blot assay를 통해 iNOS, COX-2 및 MAP kinase (p38, ERK1/2, JNK)와 NF-κB 세포내 신호전달 경로에서의 단백질 발현을 조사한 결과 이들의 발현이 유의하게 억제됨을 확인하였다. 본 연구에서는 이러한 결과를 근거하여 HCW0811가 다양한 염증 인자를 표적으로 하는 피부 면역 질환을 포함한 염증성 질환의 예방과 치료를 위한 항염증 기능성 화장품 및 식품소재로의 개발가능성을 제시한다.

• 생명과학회지
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• 제32권7호
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• pp.491-500
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• 2022

HK표고버섯균사체(HK shiitake mushroom mycelium, HKSMM)는 간 건강 개별 인정 건강기능식품이다. LPS로 활성화된 RAW 264.7 세포에서 HKSMM50 (HKSMM의 50% ethanol 수용액 추출물)의 항염증효과를 연구하였다. AHCC는 positive control로 사용하였다. LPS로 활성화된 RAW 264.7 세포에 HKSMM50 및 AHCC를 처리(0, 20, 100, 500 ㎍/ml)하고 24시간 배양하여 배양물의 염증 관련 인자는 ELISA kits로, 세포에 함유된 iNOS와 COX-2 protein 발현은 Western blotting으로 측정하였다. HKSMM50는 LPS 처리에 비해 농도 의존적으로 NF-κB 함량을 낮추었고, iNOS와 COX-2 protein 발현을 억제하여 NO와 PGE₂ 함량을 낮추었다. 더불어 HKSMM50는 LPS 처리에 비해 IL-1β, TNF-α, IL-4 및 IL-6의 함량을 낮추었으나 SOD와 CAT의 활성은 증가시켰다. AHCC도 HKSSM50 처리와 비슷한 효과를 나타내었다. 이 결과는 HKSMM50이 LPS로 활성화된 RAW 264.7 세포에서 NF-κB 신호전달을 억제하여 항염증효과를 나타내었으며, HKSMM은 면역기능증진에 도움을 줄 수 있는 건강기능식품원료로 사용할 수 있을 것이다.

• 대한한방부인과학회지
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• 제34권3호
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• pp.29-48
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• 2021

Objectives: Banchong-san (BCS) is a herbal formula composed of 13 korean medicinal herbs and is traditionally used to treat inflammatory diseases and pain. The object of this study was to research the antioxidant, anti-inflammatory, antipruritic and antimicrobial effects of the BCS in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Methods: In this experiment, effects of BCS on the following four were measured as follows: (1) Anti-oxidative effects were evaluated by 1,1-diphenyl-2-picryl-hydrazyl (DPPH) Radical scavenging activity, 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) Radical scavenging activity. (2) Anti-inflammatory effects were evaluated by the production amount of Reactive oxygen species (ROS), Nitric oxide (NO), Interleukin-1β (IL-1β), Interleukin 6 (IL-6), tumor necrosis factor-α (TNF-α), Prostaglandin E2 (PGE₂), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2)(the previous two are "mRNA"), extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein kinases (p38), inhibitor of nuclear factor kappa B (IκBα), nuclear factor kappa B (NF-κB) (the previous five are "Protein") in LPS-Stimulated RAW 264.7 cells. (3)Antipruritic effects were evaluated by the production amount of histamine, Leukotriene B4 (LTB4), LeukotrieneC4 (LTC4) Levels in phorbol 12-myristate 13-acetate(PMA)/ionomycin-stimulated MC/9 mast cell. (4) Anti-microbial effects were evaluated by the growth suppression of Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and Aspergillus niger. Results: The following results were obtained through each measurement: (1) DPPH Radical Scavenging Activity, ABTS Radical Scavenging Activity evoked a significant concentration-dependent increase. (2) ROS, NO, IL-1β, IL-6, TNF-α, PGE₂ production amount, iNOS, COX-2 mRNA expression were significantly reduced in the BCS extraction group compared with the control group and significantly decreased the amount of ERK, JNK, p38, NF-κB Protein expression. The amount of IκB-α Protein Expression have increased significantly. (3) The amounts of histamine, LTB4, LTC4 were significantly decreased. (4) The antibacterial efficacy, BCS inhibited the growth of Escherichia coli, Pseudomonas aeruginosa at concentrations of 5 ㎍/ml, but did not suppress the growth of staphylococcus aureus and aspergillus niger. Conclusions: The experimental results show that BCS has anti-oxidant, anti-inflammatory, antipruritic and antimicrobial properties.

• 대한본초학회지
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• 제28권2호
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• pp.67-73
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• 2013

Objective : Lithospermum Erythrorhizon (LE) has been used as an anti-bacterial and anti-inflammatory agent. However, it is unclear that LE aqueous extract could show the anti-inflammatory effects in RAW 264.7cells. The purpose of this study was to investigate the anti-inflammatory effect of aqueous extract from LE on lipopolysaccharide (LPS) - induced inflammatory response. Methods : To measure out the cytotoxicity of LE, we performed the MTT assay. To evaluate the anti-inflammatory effects of LE, we examined the inflammatory mediators such as nitric oxide (NO), prostaglandin E2 ($PGE_2$) and pro-inflammatory cytokines (tumor necrosis factor (TNF)-${\alpha}$, interleukin, (IL)-$1{\beta}$ and (IL)-6) on RAW 264.7 cells. We also examined molecular mechanisms such as mitogen-activated protein kinases (MAPKs) and nuclear factor-B (NF-${\kappa}B$) activation by western blot. Results : Aqueous Extract from LE itself did not have any cytotoxic effect in RAW 264.7 cells. Aqueous extract from LE inhibited LPS-induced productions of inflammatory mediators such as NO, $PGE_2$, and pro-inflammatory cytokines including TNF-${\alpha}$, IL-$1{\beta}$ and IL-6 in RAW 264.7cells. In addition, LE inhibited the phosphorylation of p38 kinases (p38), c-Jun $NH_2$-terminal kinase (JNK), and NF-${\kappa}B$ activation in RAW 264.7 cells. Conclusion : LE down-regulated LPS-induced production of inflammatory mediators through the inhibition of p38, JNK and NF-${\kappa}B$ activation. Taken together, these results could provide the evidence for the anti-inflammatory effects of LE. Therefore, LE may be a novel target in the management of inflammation and help to support a potential strategy for prevention and therapy of inflammatory diseases.

• 대한본초학회지
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• 제28권3호
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• pp.53-60
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• 2013

Objectives : DojukSan is known to be effective for treating a urinary diseases and stomatitis. However, there has been a lack of studies regarding the effects of Dojuksan on the inflammatory activities and effector inflammatory disease mechanism about macrophage before is not known. To elucidate the molecular mechanisms of Dojuksan water extract (DJS) on pharmacological and biochemical actions in inflammation, we examined the effect of DJS on pro-inflammatory mediators in lipopolysaccharide (LPS)-stimulated macrophages. Methods : In the present study, pro-inflammatory cytokine production was determined by performing enzyme-linked immunosorbent assay, reverse transcription polymerase chain reaction, and western blot analysis to measure the activation of MAPKs. Cells were treated with 200 ng/mL of LPS 1 h prior to the addition of DJS. Cell viability was measured by MTS assay. The investigation focused on whether DJS inhibited nitric oxide (NO) and prostaglandin E2 ($PGE_2$) productions, as well as the expressions of inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin-6 (IL-6) and mitogen-activated protein kinases (MAPKs) in LPS-stimulated RAW 264.7 cells. Results : We found that DJS inhibited LPS-induced NO, $PGE_2$ and IL-6 productions as well as the expressions of iNOS and COX-2. Furthermore, DJS suppressed the LPS-induced phosphorylation of p38 MAPK and c-Jun NH2-protein kinase (JNK). Conclusions : These results suggest that DJS has inhibitory effects on LPS-induced $PGE_2$, NO, and IL-6 production, as well as the expressions of iNOS and COX-2 in the murine macrophage. These inhibitory effects occur through blockades on the MAPKs phosphorylation.

• 대한본초학회지
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• 제28권1호
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• pp.43-50
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• 2013

Objectives : Hwanggeumjakyak-tang (huangqin shaoyao tang, HJT) has been used to treat acute enteritis in traditional oriental medicine. However, there has been a lack of studies regarding the effects of HJT on the inflammatory activities and effector inflammatory disease mechanism about macrophage before is not known. So we examined the effect of HJT water extract on pro-inflammatory mediators in lipopolysaccharide (LPS) - stimulated mouse macrophage, RAW 264.7 cells. Methods : Cells were treated with 2 ug/mL of LPS 1 h prior to the addition of HJT. Cell viability was measured by MTS assay. The production of nitric oxide (NO) was determined by reacting cultured medium with Griess reagent. The expression of cyclooxygenase-2 (COX-2), inducible NO synthase (iNOS) and mitogen-activated protein kinases (MAPKs) was investigated by Western blot, RT-PCR. The content of level of cytokines (prostaglandin (PG) $E_2$, interleukin (IL)-6, IL-12, Tumor necrosis factor-alpha (TNF-${\alpha}$) and monocyte chemoattractant protein-1 (MCP-1)) in media from LPS-stimulated Raw 264.7 cells was analyed by ELISA kit. Results : HJT inhibited the production of NO, $PGE_2$, IL-6 as well as the expressions of iNOS, COX-2 but did not inhibit the production of IL-12, TNF-${\alpha}$, MCP-1 in the murine macrophage, RAW 264.7 cells. HJT also had suppression effects of LPS-induced MAPKs activation Conclusion : These results suggest that HJT has an anti-inflammatory therapeutic potential, which may result from inhibition of MAPK phosphorylation, thereby decreasing the expression of pro-inflammatory genes.

• 대한본초학회지
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• 제32권4호
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• pp.25-32
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• 2017

Objectives : Chungsimyeonja-eum (CSYJE; Qingxinlianzi-yin in Chinese; Seishinrenshi-in in Japanese), a traditional herbal medicine, has been used for treating mouth dryness, hyperuresis. This study was designed to determine preservation period of CSYJE. We investigated the stability and biological activity of CSYJE depending on the preservation temperature and periods. Methods : CSYJE decoction was preserved for 0-6 or 12 months at room temperature (RT, $23{\pm}1^{\circ}C$) or refrigeration ($4^{\circ}C$). To evaluate the stability of CSYJE decoction, pH and dissolved solids content were measured. High-performance liquid chromatography (HPLC) analysis was performed to determine marker compounds-liquiritin apioside, liquiritin, baicalin, wogonoside, and glycyrrhizin-in CSYJE. To determine anti-inflammatory effect of CSYJE, prostaglandin $E_2$ ($PGE_2$) was measured in LPS-stimulated RAW 264.7 macrophages. Results : Both at RT and $4^{\circ}C$, pH was decreased depending on the preservation periods. There was no changes in dissolved solids content depending on the preservation temperature and periods. Both at RT and $4^{\circ}C$, the contents of liquiritin apioside and liquiritin were slightly increased at 1 month of storage. The level of baicalin was decreased time-dependently and the disappearance rate at RT is larger than at $4^{\circ}C$. CSYJE inhibited $PGE_2$ production in LPS-stimulated RAW 264.7 cells and maintained inhibitory effect by 12 months both at RT and $4^{\circ}C$. Conclusions : Based on the disappearance rate of the baicalin in CSYJE, the preservation period is recommended within 8 months for RT and 12 months for $4^{\circ}C$.

• 대한본초학회지
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• 제34권1호
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• pp.51-57
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• 2019

Objectives : Nypa fruticans Wurmb. (NF) have been used as a traditional medicine to treat inflammatory diseases in East-South Asia. However, it is largely undiscovered whether NF water extract could exhibit anti-inflammatory activities against tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$)-induced inflammatory responses on human keratinocytes, HaCaT cells. Therefore, this study was aimed to investigate the anti-inflammatory activity of NF water extract on TNF-${\alpha}$-induced inflammatory responses in HaCaT cells. Methods : To investigate the anti-inflammatory activites of NF water extract in HaCaT cells, the inflammatory model of HaCaT cells was established under a suitable concentration (10 ng/ml) of human TNF-${\alpha}$ (hTNF-${\alpha}$). HaCaT keratinocyte cells were pre-treated with NF water extract for 1 h, and then stimulated with hTNF-${\alpha}$. Then, the cells were harvested to measure the inflammatory mediators such as inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2 and prostaglandin $E_2$ ($PGE_2$), and pro-inflammatory cytokine including TNF-${\alpha}$ and interleukin (IL)-6. In addition, we examined the inhibitory mechanisms of NF, mitogen activated protein kinases (MAPKs) and inhibitory kappa B alpha ($I{\kappa}-B{\alpha}$) Results : The treatment of NF inhibited the hTNF-${\alpha}$-induced elevation of iNOS, COX-2, and $PGE_2$ in HaCaT cells. In addition, NF treatment inhibited the hTNF-${\alpha}$-induced elevation of TNF-${\alpha}$ and IL-6. Furthermore, NF treatment inhibited the activation of MAPKs but not degradation of $I{\kappa}-B{\alpha}$. Conclusions : Taken together, our result suggest that treatment of NF could inhibit the hTNF-${\alpha}$-induced inflammatory responses via deactivation of MAPKs in HaCaT cells. This study could suggest that NF could be a beneficial agent to prevent skin damage or inflammation.

• 대한본초학회지
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• 제33권6호
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• pp.35-42
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• 2018

Objectives : The aim of this study was to investigate whether the extract of Coptidis Rhizoma inhibits inflammation in chronic cold stress (CCS)-exposed mice or not. Methods : Coptidis Rhizoma extract (CRE) was made by reflux with distilled water. Male ICR mice (7 weeks old) were divided randomly into 5 groups: (1) control, (2) CCS, (3) CCS+CRE 100 mg/kg, (4) CCS+CRE 300 mg/kg, (5) CCS+CRE 1,000 mg/kg groups. Mice were orally administered once a day for 14 days starting from 1 day before CCS. Group (2)-(5) were exposed to CCS conditions that maintained at $4^{\circ}C$ for 2 h once a day for 14 days. The levels of serum cortisol and hypothalamic prostaglandin E1 (PGE1) and PGE2 were measured by enzyme-linked immunosorbent assay kit. The expression levels of several pro-inflammatory factors like heat shock protein 70 (HSP70), c-fos, and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) were measured by western blot analysis in mouse hypothalamus. Results : Oral administration of CRE 1,000 mg/kg significantly suppressed the increase of serum cortisol levels in mice exposed to CCS. CCS-exposed mice had significantly increased the expression of HSP70, c-fos, and NF-kB in hypothalamus, while CRE treatment significantly attenuated the elevation of these pro-inflammatory factors. The ratio of PGE2/PGE1 was also higher in CCS-exposed mice than control group. CRE treatment significantly reduced the increase of PGE2/PGE1 ratio induced by CCS. Conclusion : These findings suggest that Coptidis Rhizoma may work as a potential agent to modulate inflammatory responses under the condition of cold adaptation formed by CCS.

• 대한본초학회지
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• 제34권6호
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• pp.57-62
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• 2019

Objective : Gastritis is a major complication of gastrointestinal disease. Lonicera japonica is used in folk medicine to treat different diseases such as exopathogenic wind-heat, epidemic febrile diseases, sores, carbuncles and some infectious diseases. Therefore, this study examined the effects of Lonicera japonica water extract (LJE) on HCl/ethano-linduced acute gastric ulceration and anti-oxidants properties. Methods : LC-ESI-IT-TOF MS was employed for rapid identification of major compound from LJE. The antioxidant activities were evaluated through total polyphenol and flavonoid contents and radical scavenging assays and superoxide dismutase (SOD)-like activity. SD rats were randomly divided into five different groups including the normal group, ulcer group, positive group (20 kg/mg of omeprazole, ip), and experimental groups (100 kg/mg and 500 kg/mg of LJE, ip). Results : 4,5-Dicaffeoyl quinic acid, loganic acid, secologanic acid, sweroside, loganin, vogeloside were identified based on the detection of the molecular ion with those of literature data. The LJE was possessed free radical scavenging activities such as DPPH (IC₅₀=189.7 ㎍/㎖), ABTS (IC₅₀=164.5 ㎍/㎖), and SOD-like activity (IC₅₀=405.02 ㎍/㎖). Macroscopic and histological analyses showed LJE treated group were significantly reduced to an extent that it allowed leukocytes penetration of the gastric walls compared with the ulcer group. In addition, an ulcer inhibition rate and prostaglandin E₂ levels were increased in rats treated with LJE. Conclusion : The present study has demonstrated the antioxidantive and gastroprotective effect of LJE, these findings suggested that LJE has the potential for use in treatment of gastric disorders.