• 대한한의학방제학회지
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• 제20권2호
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• pp.83-92
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• 2012

본 연구를 통해서, 한(寒) 처방의 대표 처방인 황련해독탕(黃連解毒湯)과 열(熱) 처방의 대표 처방인 건강부자탕(乾薑附子湯) 모두 항염 효능을 확인할 수 있었으나, 그 작용 기전에 있어 뚜렷한 차이를 나타내었다. 이러한 차이는 한의학의 기본 이론인 한열에 대한 개념에 대한 연구의 초석이 될 수 있기를 바라며, 각 개별 약물의 효능 및 다른 처방들과 다른 기전적 실험이 추가적으로 필요할 것을 보인다.

• Biomolecules & Therapeutics
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• 제26권2호
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• pp.146-156
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• 2018

Spermidine is a naturally occurring polyamine compound that has recently emerged with anti-aging properties and suppresses inflammation and oxidation. However, its mechanisms of action on anti-inflammatory and antioxidant effects have not been fully elucidated. In this study, the potential of spermidine for reducing pro-inflammatory and oxidative effects in lipopolysaccharide (LPS)-stimulated macrophages and zebrafish was explored. Our data indicate that spermidine significantly inhibited the production of pro-inflammatory mediators such as nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$), and cytokines including tumor necrosis $factor-{\alpha}$ and $interleukin-1{\beta}$ in RAW 264.7 macrophages without any significant cytotoxicity. The protective effects of spermidine accompanied by a marked suppression in their regulatory gene expression at the transcription levels. Spermidine also attenuated the nuclear translocation of $NF-{\kappa}B$ p65 subunit and reduced LPS-induced intracellular accumulation of reactive oxygen species (ROS) in RAW 264.7 macrophages. Moreover, spermidine prevented the LPS-induced NO production and ROS accumulation in zebrafish larvae and was found to be associated with a diminished recruitment of neutrophils and macrophages. Although more work is needed to fully understand the critical role of spermidine on the inhibition of inflammation-associated migration of immune cells, our findings clearly demonstrate that spermidine may be a potential therapeutic intervention for the treatment of inflammatory and oxidative disorders.

• Molecules and Cells
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• 제41권8호
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• pp.742-752
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• 2018

Parkinson's disease (PD) is a neurodegenerative disease characterized by progressive degeneration of dopaminergic (DAergic) neurons, particularly in the substantia nigra (SN). Although circadian dysfunction has been suggested as one of the pathophysiological risk factors for PD, the exact molecular link between the circadian clock and PD remains largely unclear. We have recently demonstrated that $REV-ERB{\alpha}$, a circadian nuclear receptor, serves as a key molecular link between the circadian and DAergic systems. It competitively cooperates with NURR1, another nuclear receptor required for the optimal development and function of DA neurons, to control DAergic gene transcription. Considering our previous findings, we hypothesize that $REV-ERB{\alpha}$ may have a role in the onset and/or progression of PD. In the present study, we therefore aimed to elucidate whether genetic abrogation of $REV-ERB{\alpha}$ affects PD-related phenotypes in a mouse model of PD produced by a unilateral injection of 6-hydroxydopamine (6-OHDA) into the dorsal striatum. $REV-ERB{\alpha}$ deficiency significantly exacerbated 6-OHDA-induced motor deficits as well as DAergic neuronal loss in the vertebral midbrain including the SN and the ventral tegmental area. The exacerbated DAergic degeneration likely involves neuroinflammation-mediated neurotoxicity. The $REV-erb{\alpha}$ knockout mice showed prolonged microglial activation in the SN along with the over-production of interleukin $1{\beta}$, a pro-inflammatory cytokine, in response to 6-OHDA. In conclusion, the present study demonstrates for the first time that genetic abrogation of $REV-ERB{\alpha}$ can increase vulnerability of DAergic neurons to neurotoxic insults, such as 6-OHDA, thereby implying that its normal function may be beneficial for maintaining DAergic neuron populations during PD progression.

• 대한예방한의학회지
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• 제16권3호
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• pp.139-153
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• 2012

Objectives : NK cells are spontaneously cytotoxic lymphocytes. These are not only important parts in the first line of defence against bacterial and viral infections of outside, but they may also play a critical role in chronic viral diseases. NK cells kill their targets spontaneously, without the need for prior sensitization and class I MHC restriction by the regulation of cytolytic functions and secretion of a variety of cytokines, such as interleukin-12(IL-12), MCP-1, IL-6, TNF-${\alpha}$, IFN-${\gamma}$. In addition, macrophage and NK cells cooperate through the production of cell mediates. These cooperation and modulation are one of major factors to prevent for evading immune surveillance of cancer. Hence, it could be assumed that if any candidate to enhance activities of macrophage and NK cell, it is considered as a potentially useful agents against cancer. Methods : In our study, to investigate effect of fermented soybean extracts by Lactic acid bacteria (SFE, soybean fermented extracts) work on intestinal immune cell to maintain general immune modulating and anti-cancer activity. We analyzed NK cytotoxicity assay and gene expressions of cytokine related with macrophage and NK cell activity. Results : In vitro experiment, SFE was verified as safety material for cell toxicicty to tumor cell strain without any toxicity of tumor growth inhibition and various cell strain. Effects of macrophage activity stimulating directly by SFE measured induced cytokine. The studies showed that IL-12 production by stimulation of SFE depended on concentration from 0.16mg/mL to 0.63mg/mL with non toxicity to cell, and it was the best activity at 0.63mg/mL. Besides, the effective concentration of SFE producing TNF-${\alpha}$ is similar to IL-12, but it was the best activity at 1.25mg/mL. The level of MCP-1, IL-6 and IFN-${\gamma}$ depended on concentration from 0.16mg/mL to 10mg/mL, IFN-${\gamma}$ showed the best activity at the effective concentration of 0.63mg/mL. With the result of NK cell activity measurement, the spleen cell of mouse injected SFE had 1.5 times higher killing effect than non injected cell. Conclusions : The result of this studies is that Soybean fermetated extracts(SFE) has possibility to immune aided material for the function not only inhibition of microbial infection to macrophage but also activity of adaption immune and cellular immune system.

• 한국버섯학회지
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• 제19권3호
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• pp.176-183
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• 2021

본 연구는 국내 참나무 생육지에서 채집된 송로버섯의 일종인 T. himalayense 자실체 추출물(TH)의 항염증 활성을 확인하였다. LPS로 유도된 RAW 264.7 대식세포를 대상으로 한 염증 억제 실험에서 TH는 100 ㎍/ml 이하의 농도에서 세포독성을 보이지 않았으며 LPS에 의해 증가된 NO와 PGE₂의 생성을 농도 의존적으로 억제하였다. Western blot 분석 결과로 볼 때, TH의 항염증 활성은 iNOS와 COX-2 유전자의 발현억제에 의해 NO와 PGE₂의 생성이 감소된 것으로부터 유발된 것임을 입증한다. 또한, TH가 대식세포에 의해 생산 및 분비되는 cytokine IL-1β와 IL-6의 생성을 효과적으로 저해함을 확인하였다. T. himalayense는 다양한 염증 매개체들의 과발현을 효과적으로 억제하며, 이를 표적으로 하는 항염증 관련 식품 및 의약품에 다양하게 활용 될 수 있을 것으로 기대된다. 추후 염증 예방 물질로 적용 될 수 있는 가능성을 높이기 위하여 염증성 작용 기전에 대한 자세한 연구가 필요할 것으로 사료된다.

• Biomolecules & Therapeutics
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• 제27권4호
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• pp.414-422
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• 2019

There is accumulating evidence that microRNAs are emerging as pivotal regulators in the development and progression of neuropathic pain. MicroRNA-15a/16 (miR-15a/16) have been reported to play an important role in various diseases and inflammation response processes. However, whether miR-15a/16 participates in the regulation of neuroinflammation and neuropathic pain development remains unknown. In this study, we established a mouse model of neuropathic pain by chronic constriction injury (CCI) of the sciatic nerves. Our results showed that both miR-15a and miR-16 expression was significantly upregulated in the spinal cord of CCI rats. Downregulation of the expression of miR-15a and miR-16 by intrathecal injection of a specific inhibitor significantly attenuated the mechanical allodynia and thermal hyperalgesia of CCI rats. Furthermore, inhibition of miR-15a and miR-16 downregulated the expression of interleukin-$1{\beta}$ and tumor-necrosis factor-${\alpha}$ in the spinal cord of CCI rats. Bioinformatic analysis predicted that G protein-coupled receptor kinase 2 (GRK2), an important regulator in neuropathic pain and inflammation, was a potential target gene of miR-15a and miR-16. Inhibition of miR-15a and miR-16 markedly increased the expression of GRK2 while downregulating the activation of p38 mitogen-activated protein kinase and $NF-{\kappa}B$ in CCI rats. Notably, the silencing of GRK2 significantly reversed the inhibitory effects of miR-15a/16 inhibition in neuropathic pain. In conclusion, our results suggest that inhibition of miR-15a/16 expression alleviates neuropathic pain development by targeting GRK2. These findings provide novel insights into the molecular pathogenesis of neuropathic pain and suggest potential therapeutic targets for preventing neuropathic pain development.

• Animal Bioscience
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• 제34권7호
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• pp.1169-1180
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• 2021

Objective: This research aimed to study the effects of Artemisia argyi flavonoids (AAF) supplemented in diets on the growth performance and immune function of broiler chickens challenged with lipopolysaccharide (LPS). Methods: A total of one hundred and ninety-two 1-d-old broiler chicks were assigned into 4 treatment groups, which were, respectively, fed a basal diet (control), fed a diet with 750 mg/kg AAF, fed a basal diet, and challenged with LPS, fed a diet with 750 mg/kg AAF, and challenged with LPS. Each treatment had six pens with 8 chicks per pen. On days 14, 16, 18, 20 (stress phase I) and 28, 30, 32, 34 (stress phase II), broilers were injected with LPS (500 ㎍/kg body weight) or an equivalent amount of saline. Results: The results demonstrated that dietary AAF significantly improved the body weight (d 21) and alleviated the decrease of average daily gain in broilers challenged with LPS on d 21 and d 35 (p<0.05). Dietary AAF increased bursa fabricius index, and dramatically attenuated the elevation of spleen index caused by LPS on d 35 (p<0.05). Furthermore, serum interleukin-6 (IL-6) concentration decreased with AAF supplementation on d 21 (p<0.05). Diet treatment and LPS challenge exhibited a significant interaction for the concentration of IL-1β (d 21) and IL-6 (d 35) in serum (p<0.05). Additionally, AAF supplementation mitigated the increase of IL-1β, IL-6 in liver and spleen induced by LPS on d 21 and 35 (p<0.05). This study also showed that AAF supplementation significantly reduced the expression of IL-1β (d 21) and nuclear transcription factor kappa-B p65 (d 21 and 35) in liver (p<0.05), and dietary AAF and LPS treatment exhibited significant interaction for the gene expression of IL-6 (d 21), toll like receptor 4 (d 35) and myeloid differentiation factor 88 (d 35) in spleen (p<0.05). Conclusion: In conclusion, AAF could be used as a potential natural immunomodulator to improve growth performance and alleviate immune stress in broilers challenged with LPS.

• 대한본초학회지
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• 제37권3호
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• pp.9-19
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• 2022

Objectives : Angelicae Gigantis Radix (AG) is a plant of the Ranunculus family. AG have been reported to have various pharmacological effects on human health which include uterine growth promotion, anti-inflammatory, analgesic, and immune enhancement. However, research on dermatitis disease is insufficient. Therefore, we investigated the effects of AG on tumor necrosis factor-α (TNF-α)/interferon-γ (IFN-γ) stimulated HaCaT cell. Methods : To investigate the effect of AG on HaCaT cell, HaCaT cells were pre-treated with AG for 1 hour and then stimulated with TNF-α/IFN-γ. After 24 hours, media and cells were harvested to analyze the inflammatory mediators. Concentration of human interleukin-1beta (IL-1β), monocyte chemoattractant protein-1 (MCP-1), granulocyte-macrophage colony-stimulating factor (GM-CSF), and TNF-α in the media were assessed by ELISA. mRNA expression of human thymus and activation-regulated chemokine (TARC), IL-6, and IL-8 were analyzed by RT-PCR. Additionally, the mechanisms of mitogen-activated protein kinases (MAPKs) and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling pathway were investigated by Western blot. Results : The treatment of AG inhibited gene expression levels of IL-6, IL-8, and TARC and protein expression levels of IL-1β, MCP-1, and GM-CSF. Also, AG significantly reduced extracellular signal-regulated kinase (ERK) phosphorylation and NF-κB translocation in TNF-α/IFN-γ stimulated HaCaT cell. Conclusions : Taken together, these results demonstrate that AG can alleviate inflammatory diseases such as atopic dermatitis by regulating the expression of inflammatory cytokines. Also, it suggest that AG may a promising candidate drug for the treatment of inflammatory disease such as atopic dermatitis.

• 농업생명과학연구
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• 제50권2호
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• pp.139-150
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• 2016

말뼈추출물은 다양한 골질환의 예방과 치료에 탁월한 효능이 있다고 이전에 보고되었다. 하지만 말뼈추출물의 다른 약리학적 효능에 대해서는 아직 자세히 밝혀지지 않고있다. 본 연구에서는 말뼈추출물이 중요한 항산화 인자인 hemeoxygenase-1(HO-1)의 발현을 상승시킬 수 있는지, 만약 발현이 증가한다면 HO-1의 상향 조절이 대식세포에서 항염증 효과를 매개할 수 있는지에 관하여 조사하였다. 이를 위해서 nitric oxide(NO) 농도측정, 세포 생존능 측정, DPPH 라디칼 소거능 검사를 시행하였다. 또한 염증성 사이토카인 유전자 발현과 단백질 발현을 측정하기 위해 real time PCR과 Western blotting을 시행하였다. 말뼈추출물은 lipopolysaccharide(LPS, 0.1㎍/ml)로 자극한 대식세포주인 RAW264.7 세포에서 어떠한 세포독성 없이 NO의 생성을 유의성 있게 억제하였으며 inducible nitric oxide(iNOS)와 cyclooxygenase 2(COX-2)의 발현을 억제하였다. 뿐만 아니라 말뼈 추출물은 염증성 사이토카인인 tumor necrosis factor(TNF)-α와 interleukin(IL)-1β의 발현을 억제하였으며 ERK, JNK 및 p-38 MAPK의 단백질 인산화를 억제하였다. 그리고 말뼈추출물은 HO-1과 NF-E2-related factor-2(Nrf-2) 의 발현을 증가시켰고 이것은 말뼈추출물이 가지고 있는 항 염증효과를 매개할 수 있는 것으로 보인다. 즉, 말뼈추출물이 HO-1의 발현을 상향 조절한 반면 ERK1/2의 신호전달 경로에 손상을 주는 것으로 확인되었으며 이러한 말뼈추출물의 효과가 최종적으로 세포손상과 세포의 과산화 자극으로부터 세포를 보호 할 수 있는 것으로 사료된다.

• 한국어병학회지
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• 제36권2호
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• pp.337-348
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• 2023

본 연구에서는 넙치의 해독 과정에서 amprolium hydrochloride의 영향을 평가하기 위해 수행되었다. 이전 연구에서 보고된 amprolium의 LD₅₀ 값을 이용하여 두 가지 실험을 진행하였다. 첫 번째는 30마리의 넙치를 5개의 대조군 및 실험군으로 나누었고 4, 8, 16, 32 mg/kg 용량의 amprolium을 근육 내 주사 투여하였다. 주사 후 8, 24, 48 시간에 간과 신장을 적출하여 약물 대사 효소와 전염증성 사이토카인 유전자의 발현을 분석하였다. 32 mg/kg 용량의 실험군에서 IL-1β mRNA의 높은 발현을 확인하였고, CYP1A는 이와 반대의 결과를 보였으며, 간에서 UGT와 GST mRNA의 발현은 유의하게 감소하는 것을 확인하였다. 또한 신장에서 amprolium 주사 투여 후 약물 대사 효소와 사이토카인 유전자의 억제가 관찰되었다. 또 다른 실험에서는 4, 8, 16, 32 mg/kg과 60, 80, 100, 120 mg/kg의 용량을 설정하여 근육 내 주사 투여하였다. 주사를 완료하고 6일 후 간을 적출하여 유전자의 발현을 확인하였다. IL-1β의 발현은 4 mg/kg 용량 실험군에서 유의적으로 매우 높은 발현을 보였다. GST의 mRNA 발현 또한 4 mg/kg 용량 실험군에서 높은 발현을 보였다. 결론적으로 우리의 결과는 amprolium이 가축 산업의 가장 안전한 합성 항콕시듐 약물 중 하나로 간주되지만 넙치의 간접 또는 직접적인 물리적 또는 생물학적 독성을 유발하는 것으로 판단된다.