• Restorative Dentistry and Endodontics
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• 제25권4호
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• pp.606-618
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• 2000

Dental pulp infection is most commonly caused by extensive dental caries, and some bacterial species invade root canals; bacterial components and products are thought to be associated with the pathogenesis of periapical periodontitis. A principle driving force behind pulpal disease response appears to lie in the host immune system's to bacteria and their products. We examined the production of interleukin $1{\beta}$ (IL-$1{\beta}$) and tumor necrosis factor ${\alpha}$(TNF-${\alpha}$) from human peripheral mononuclear cells, lymphocytes and monocytes stimulated by heat-killed Acitnobacillus actinomycetemcomitans (ATCC 29523), Porphyromonas gingivalis (ATCC 33277) and Prevotella intermedia (ATCC 25611), and also by their sonicated bacterial extracts (SBE), respectively. The effects of three strains of heat-killed bacteria and their SBEs on the morphology of cultured blood cell lines HL-60 (KCLB 10240) and J774A.1 (KCLB 40067) were observed under the inverted microscope. Ultrastructural changes of J774A.1 exposed to heat-killed P. intermedia and its SBE were investigated using transmission electron microscopy. Production of IL-$1{\beta}$ was reduced in human peripheral mononuclear cells after stimulation by sonic bacterial extracts of A. actinomycetemcomitans, P. gingivalis, and P. intermedia. Heat-killed and sonic extract of P. gingivalis inhibited the production of TNF-${\alpha}$ in peripheral mononuclear cells. Production of TNF-${\alpha}$ was inhibited in peripheral monocytes after stimulation by sonic extracts of A. actinomycetemcomitans, P. gingivalis, and P. intermedia. HL-60 and J 774A.1 cells showed granular degeneration after treatment with heat-killed and sonic extracts of A. actinomycetemcomitans, P. gingivalis, and P. intermedia Chromatin margination and shrinkage were observed in 774A.1 treated with heat-killed P. intermedia. Cell wall structure and organelles were destroyed and vacuoles were formed in cytoplasm in J774A.1 treated with P. intermedia sonic extract. These results suggest that A actinomycetemcomitans, P gingivalis and P intermedia may have an important role in the formation and progression of pulpal diseases via both modulation of production of IL-$1{\beta}$ and TNF-${\alpha}$ from blood mononuclear cells and cytopathic effects.

• Journal of Microbiology and Biotechnology
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• 제16권10호
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• pp.1544-1553
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• 2006

The halophilic enterobacteria, Enterobacteria cancerogenus, was isolated from the intestines of the fusiform fish (Trachurus japonicus) to yield a protein-like material termed PLM-f74. PLM-f74 was characterized by strong inhibition ratios to angiogenesis (82.8% at the concentration of $18.5{\mu}g/ml$) and elevated antioxidative capacities with low toxicity. The PLM-f74 is a glycoprotein comprised of saccharides and amino acids. PLM-f74 inhibited cell adhesion that non-activated U937 monocytic cell adhesion to HUVECs activated with $IL-1{\beta}$ by 78.0%, and the adherence of U937 cells treated with the PLM-f74 and stimulated with $IL-1{\beta}$ to unstimulated HUVECs decreased by 102%. When both cell types were pretreated with PLM-f74, the adhesion of U937 cells to $IL-1{\beta}$-stimulated HUVECs was completely suppressed by 121% at a concentration of $18.5{\mu}g/ml$. PLM-f74 blocked signal pathways from VEGFR2, PI3K, ${\beta}$-catenin, and VE-cadherin to NF-kB, based on western bolt analysis. It also inhibited IL-l-stimulated HUVEC expression of the adhesion molecules, ICAM-l by 40%, VCAM-l by 60%, and E-selectin by 70% at the same concentration noted above. New anti-angiogenic and anti-cell adhesion materials showing elevated antioxidative capacities, and non-toxicity may be expected from these results.

• 생명과학회지
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• 제21권8호
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• pp.1176-1182
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• 2011

본 연구는 고지방식이로 유도된 비만쥐를 대상으로 저지방식이와 중강도 운동이 비장세포의 전염증성 사이토카인 발현에 미치는 영향을 보고자 하였다. 4 주령의 C57BL/6 마우스를 대상으로 고지방식이(45% fat)를 5 주간 처치하여 비만을 유도한 후, 저지방식이(10% fat)로 전환한 후 통제군(운동미실시, n=10)과 운동군(중강도운동실시, n=10)으로 나누어 8 주간 트레드밀을 이용하여 중강도 운동을 주 5회, 1 일 30-60분 실시하였다. 운동종료후 비장세포를 분리하여 Concanavalin A (10 ug/ml)로 24시간 자극 후 전염증성 사이토카인인 IL-1${\beta}$, IL-6, TNF-${\alpha}$를 Bio-Plex를 활용하여 측정하였다. 자료분석은 독립 t 검증을 실시 하였으며, 유의수준은 p<0.05 수준으로 하였다. 비장세포에서 발현된 전염증성 사이토카인인 IL-1${\beta}$, IL-6, TNF-${\alpha}$는 저지방식이를 단독처치한 통제군에 비하여 중강도 운동을 병행실시한 운동군에서 유의하게 감소한 결과(p<0.01)를 나타내어 고지방식이로 유도된 비만쥐에서 저지방식이의 단독처치보다 중강도 운동의 병행 처치가 전염증 사이토카인 감소에 긍정적인 영향을 미치는 것으로 사료된다.

• 한국식품영양학회지
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• 제29권3호
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• pp.431-437
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• 2016

본 연구는 전통약재로 주로 사용되는 구절초에서 분리한 다당류(CZPS)가 선천 및 적응면역에서 중추적인 역할을 수행하는 대식세포의 활성화에 미치는 영향에 관하여 관찰하였다. 구절초에서 분리한 다당류를 마우스 유래 대식세포주인 RAW 264.7 cell에 처리하였을 때, iNOS의 발현을 조절하여 NO의 생성이 증가되었으며, 또한 면역활성 물질인 TNF-${\alpha}$, IL-$1{\beta}$ 및 IL-6 등의 cytokine의 분비능이 증가되었으며, 이러한 면역활성 매개자인 NO 및 cytokine의 증가의 원인에 관한 정확한 면역세포내 신호전달에 관하여 알아본 결과, CZPS 처리는 MAPKs(ERK, p38)의 인산화를 촉진시키며, 이로 인한 전사인자인 NF-${\kappa}B$의 인산화를 증가시켜, 대식세포의 활성화를 유도시키는 것으로 관찰되었다.

• Tuberculosis and Respiratory Diseases
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• 제44권4호
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• pp.729-741
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• 1997

연구배경 : 저자들은 최근 결핵균의 자극에 의해 peripheral blood monocytes (PBM)에서 IL-$1{\beta}$와 $TNF{\alpha}$가 생성되고, 기도상피세포에서는 IL-8과 RANTES와 같은 chemokine이 생성된다는 사실을 밝힌 바 있다. 이와같은 사실은 기도상피세포가 IL-8과 RANTES와 같은 chemokine을 분비함으로써 결핵의 병인에 능동적으로 참여함을 시사한다고 하겠다. 기도상피세포에서 발현하는 chemokine들이 결핵의 병태생리에 관여한다면, 폐결핵을 일으키는 가진 균주인 H37Rv와 폐결핵을 일으키지 않는 균주인 H37Ra로 자극하였을때 기도상피세포에서 분비되는 chemokine의 성상이 다를 것이라는 가정이 가능하다. 실제로 독성을 가지고 있는 Erdman 균주와 독성을 가지고 있지 않는 H37Ra의 lipoarabinomannan(LAM) 의 구조가 다르고, 이 LAM으로 대식세포를 자극하였을때 H37Ra LAM은 대식세포에서 $TNF{\alpha}$를 생성시키지만 H37Rv LAM은 $TNF{\alpha}$를 생성시키지 못하고 이와같은 차이가 결핵균에 대한 생체의 방어기전을 설명한다는 보고가 있다. 본 연구는 폐결핵의 병태생리에서 기도상피세포의 역할을 규명하고자, 독성이 있는 균주인 H37Rv와 독성이 없는 균주인 H37Ra를 사용하여 결핵균의 독성여부에 따른 기도상피세포에서 분비되는 chemokine의 차이를 연구하고자 하였다. 방 법 : 정상인에서 채취한 말초혈액에서 말초혈액 단핵세포(PBM)를 분리하여, sonicated H37Rv, H37Ra($5{\times}10^5$ bacilli, A TCC) 또는 LPS($10{\mu}l/ml$)로 자극하면서 24시간 동안 배양하였다. 또한 PBM에 대한 interferon gamma($IFN{\gamma}$)의 영향을 평가하고자 PBM를 결핵균으로 자극하기 한 시간전에 $IFN{\gamma}$(10ng/ml, R & D) 로 전처치한 군과 $IFN{\gamma}$ 전처치 없이 자극한 군을 두었다. 24시간후 상층액을 채취하여 상층액의 일부는 $-70^{\circ}C$에서 보관하였고 이후 ELISA kit(R & D)를 이용하여 $TNF{\alpha}$와 $IL-1{\beta}$의 농도를 측정하였다. 배양된 PBM의 상충액(1 : 2 희석액 )으로 배양된 A549 세포를 다시 자극하면서 24시간 동안 배양하였다. 24시간 배양후, 상층액은 IL-8, RANTES의 측정을 위하여 $-70^{\circ}C$에서 보관하고, 남아있는 A549 세포에서 tRNA를 추출하였다. 배양 상층액에서는 ELISA kit(R & D)를 이용하여 IL-8, RANTES의 농도를 측정하였고, 추출한 tRNA는 Northern blot analysis를 이용하여 IL-8, RANTES의 mRNA의 양을 측정 정량하였다. 결 과 : 말초혈액 단핵세포를 LPS, H37Rv, 또는 H37Ra로 자극하였을 때 대조군에 바하여 $TNF{\alpha}$와 IL-$1{\beta}$의 생성이 읖미 있게 증가하였다. 독성이 없는 H37Ra로 자극하였을 때가 독성이 있는 H37Rv로 자극하였을 때보다 $TNF{\alpha}$와 IL-$1{\beta}$의 생성이 많았으나 통계적인 유의성은 없었다. $IFN{\gamma}$로 전처치하였을 때 $TNF{\alpha}$와 IL-$1{\beta}$의 생성이 전처치하지 않은 군에 비해 증가하였으나 H37Rv군과 H37Ra군 사이에 통계적으로 유의한 차이는 없었다. A549 세포를 결핵균이나 LPS로 자극한 말초혈액 단핵세포 배양액으로 자극하였을 때, RANTES와 IL-8 의 유전자 발현이 대조군에 비하여 의미있게 증가하였다(p<0.001). 또한 H37Ra 배양액으로 자극한 군에서 H37Rv 배양액으로 자극한 군에 비하여 RANTES와 IL-8의 유전자 발현이 의미있게 증가하였다 (p<0.05). A549 세포를 결핵균이나 LPS로 자극한 말초혈액 단핵세포 배양액으로 자극하였을 때 RANTES와 IL-8의 생성이 대조군에 비하여 의미있게 증가하였다. 그러나 유전자 발현과는 달리, H37Ra 배양액으로 자극한 군에서 H37Rv 배양액으로 자극한 군에 비하여 RANTES와 IL-8의 생성이 증가하였지만 통계적인 유의성은 없었다. 결 론 : 기도상피세포는 폐결핵에서 RANTES와 IL-8과 같은 강력한 chemokine을 분비하여 염증반응을 증폭시킴으로써 폐결핵의 병인에 능동적으로 참여함을 추정할 수 있었다. 그러나 말초혈액 단핵세포에서의 $TNF{\alpha}$와 IL-$1{\beta}$의 생성은 결핵균주의 독성여부에 따라 차이가 없었고, A549 세포에서도 RANTES와 IL-8의 유전자 발현은 H37Ra 에서 유의하게 증가하였으나 RANTES와 IL-8의 생성은 유의한 차이가 없어 결핵균의 독성 여부가 균주에 따른 생체의 면역반응즉 chemokine 생성의 차이 때문이라는 본 연구의 가설을 증명할 수는 없었다.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2019년도 춘계학술대회
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• pp.94-94
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• 2019

In this study, we elucidated the anti-inflammatory mechanisms of leaves extracts from Rodgersia podophylla (RPL) in RAW264.7 cells. RP-L significantly inhibited the production of the proinflammatory mediators such as NO, iNOS, IL-$1{\beta}$ and IL-6 in LPS-stimulated RAW264.7 cells. RPL increased HO-1 expression in RAW264.7 cells, and the inhibition of HO-1 by ZnPP reduced the inhibitory effect of RPL against LPS-induced NO production in RAW264.7 cells. Inhibition of p38, ROS and $GSK3{\beta}$ attenuated RPL-mediated HO-1 expression. Inhibition of ROS inhibited p38 phosphorylation and $GSK3{\beta}$ expression induced by RPL. In addition, inhibition of $GSK3{\beta}$ blocked RPL-mediated p38 phosphorylation. RPL induced nuclear accumulation of Nrf2, and Inhibition of p38, ROS and $GSK3{\beta}$ abolished RPL-mediated nuclear accumulation of Nrf2. Furthermore, RPL blocked LPS-induced degradation of $I{\kappa}B-{\alpha}$ and nuclear accumulation of p65. RP-L also attenuated LPS-induced phosphorylation of ERK1/2 and p38. Our results suggest that RPL exerts potential antiinflammatory activity by activating ROS/$GSK3{\beta}$/p38/Nrf2/HO-1 signaling and inhibiting NF-${\kappa}B$ and MAPK signaling in RAW264.7 cells. These findings suggest that RPL may have great potential for the development of anti-inflammatory drug.

• Korean Journal of Acupuncture
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• 제29권1호
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• pp.37-46
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• 2012

Objectives : The gnarl of Pinus densiflora, called Songjeol in Korea, has been used as a medicinal herb for the treatment of inflammatory-related diseases such as arthralgia, myalgia and bruise. However, the molecular actions and mechanisms have not been clearly investigated. The aim of this study was to clarify the anti-inflammatory activity of Pinus densiflora gnarl pharmacopuncture (PDGP) in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Methods : Cytotoxicity was assessed by XTT assay. The amount of nitric oxide (NO) production was determined by nitrite assay. The mRNA expressions of interleukin-$1{\beta}$ (IL-$1{\beta}$), interleukin-6 (IL-6), cyclooxygenase-2 (COX-2) and heme oxygenase-1 (HO-1) were analyzed by RT-PCR. Reactive oxidative species (ROS) generation was measured using the fluorescence microscopy. In addition, inducible nitric oxide synthase (iNOS) and redox factor-1 (Ref-1) protein expressions were detected by Western blotting. Results : PDGP inhibited NO production and ROS generation in LPS-stimulated RAW264.7 cells. At the mRNA level, PDGP suppressed IL-$1{\beta}$, IL-6 and COX-2 expression. On the other hand, PDGP induced HO-1 mRNA expression. Furthermore, PDGP suppressed iNOS and Ref-1 protein expression. Conclusions : This result suggests that PDGP can act as a suppressor agent on NO and iNOS through induction of HO-1, and play an useful role in blocking inflammatory responses.

• Biomolecules & Therapeutics
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• 제21권5호
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• pp.398-404
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• 2013

During a screening program to search the anticolitic herbal medicines, 80% ethanol extract of the rhizome of Anemarrhena asphodeloides (AA) was found to potently inhibit the expression of proinflammatory cytokines TNF-${\alpha}$ and IL-1${\beta}$, as well as the activation of NF-${\kappa}B$ in LPS-stimulated colonic macrophages, followed by that of the rhizome of C. chinensis (CC). AA also potently inhibited TNBS-induced colitic markers, shortening of the colon and increase of macroscopic score, myeloperoxidase activity, TNF-${\alpha}$, IL-1${\beta}$, and IL-6, in mice. The synergistic effect of CC against the anticolitic effect of AA was investigated. CC synergistically inhibited the anticolitic effect of AA. AC-mix (AA+CC, 1:1) potently inhibited them. AC-mix also inhibited the activation of NF-${\kappa}B$, as well as the expression of TNF-${\alpha}$, IL-1${\beta}$, IL-6, iNOS and COX-2. The effects of AC-mix against oxazolone-induced colitis were investigated in mice. AC-mix also potently inhibited oxazolone-induced inflammatory markers, colon shortening, macroscopic score, myeloperoxidase activity, NF-${\kappa}B$ activation and proinflammatory cytokines. Overall, the anti-colitic effect of AC-mix was superior to that of mesalazine. Based on these findings, AC-mix may improve colitis by inhibiting NF-${\kappa}B$ activation.

• 동의생리병리학회지
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• 제31권4호
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• pp.220-225
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• 2017

Lithospermi Radix (LR) is known to have an anti-inflammatory effect. However, the mechanisms are not well known. In this study, LPS-induced mouse RAW 264.7 macrophage cells were treated with LR to investigate the time-dependent inflammation response of LR. RAW 264.7 cells were treated with various concentrations of LR for 24 hours, followed by MTS assay. Cell viability was increased at all experimental concentrations. The mRNA expression levels of $IL-1{\beta}$, $TNF-{\alpha}$ and iNOS were increased by treatment of RAW 264.7 cells with LR at a concentration of $200{\mu}g/ml$ for 6 hours and 24 hours. Treatment of LR with $200{\mu}g/ml$ concentration for 6 hours promoted mRNA expression levels of $IL-1{\beta}$, $TNF-{\alpha}$ and iNOS in LPS-induced RAW 264.7 cells. However, $IL-1{\beta}$, $TNF-{\alpha}$ and iNOS mRNA expression was suppressed by treatment of LR with $200{\mu}g/ml$ concentration for 24 hours in LPS-induced RAW 264.7 cells. These results suggest that the effect on inflammation of LR is promptly promoted and then to rapidly alleviate the inflammatory reaction. This study proposes that the time-dependent activities of herbal medicine is a very important factor in analyzing the anti-inflammatory effect of various herbal medicines including LR.

• 한방재활의학과학회지
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• 제23권4호
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• pp.39-57
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• 2013

Objectives The purpose of this study is to prove the effect of Kyejakjimotangkami(KMK) on osteoarthritis. Methods We checked antioxidant activity and measured production of $IL-1{\beta}$, IL-6, TNF-${\alpha}$ in RAW 264.7 cell after treat by KMK. Then we measured hind paw weight of Wister Rat with arthritis induced by MIA after KMK oral administration, checked Prostaglandin E2, IL-$1{\beta}$, IL-6, TNF-${\alpha}$, Osteocalcin, TIMP-1, MMP-9, LTB-4 in serum, ran histopathological test and ${\mu}CT$-arthrography. Results 1. DPPH radical Scavenging was increased depend on concentration of KMK ethanol extract in RAW 264.7 cell. 2. Production of NO was significantly decreased by KMK ethanol extract on concentration of $200{\mu}g/ml$ in RAW 264.7 cell. 3. Production of IL-$1{\beta}$ was significantly decreased by KMK ethanol extract on concentration of $200{\mu}g/ml$. And Production of IL-6, TNF-${\alpha}$ were significantly decreased KMK ethanol extract of every concentration in RAW 264.7 cell. 4. Result of checking hind paw weight when administered KMK ethanol extract to Wister Rat with arthritis induced by MIA was significantly higher than control group and similar to normal group. 5. Production of Prostaglandin E2, IL-$1{\beta}$, Osteocalcin, TIMP-1, MMP-9 and LTB-4 in serum was significantly decreased by KMK ethanol extract after administerd to Wister Rat with arthritis induced by MIA. 6. In Hematoxylin & Eosin staining and Safranin-O staining, we could find inflammation of synovial cell, infiltration of macrophage and granulocyte and degeneration of cartilage and bone were decreased in comparison with control group. 7. When checked cartilage volume to examine degree of cartilage degeneration using ${\mu}CT$-arthrography, volume of cartilage was increased in comparison with control group. Conclusions Comparison of the results for this study showed that KMK ethanol extract have anti-inflammatory effectiveness and can protect cartilage and bone. So we expect that KMK can be used as a effective drugs for osteoarthritis.