• 대한한방소아과학회지
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• 제28권2호
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• pp.23-39
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• 2014

Objectives PRAL (Prunus armniaca Linne Var) is a herbal formula in Oriental Medicine, known for its anti-inflammatory and anti-allergenic properties. However, its mechanism of action and the cellular targets have not yet been found enough. The purpose of this study is to investigate the effects of PRAL on Th2 cytokines expression in MC/9 mast cells. Methods The effect of PRAL was analyzed by ELISA, Real-time PCR, Western blot in MC/9 mast cells. mRNA levels of GM-CSF, IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$ were analyzed with Real-time PCR. Levels of IL-13, MIP-$1{\alpha}$ were measured using enzyme-linked immunosorbent assays (ELISA). NFAT, AP-1 and NF-${\kappa}B$ p65 were examined by Western blot analysis. Results PRAL inhibited GM-CSF, IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$ mRNA expression in a dose dependent manner. GM-CSF, IL-4, IL-5 mRNA expression were inhibited significantly in comparison to DNP-IgE control group at concentration of 100 ${\mu}g/ml$ and IL-6, IL-13, TNF-${\alpha}$ mRNA expression were inhibited at concentration of 50 ${\mu}g/ml$, 100 ${\mu}g/ml$. PRAL also inhibited the IL-13, MIP-$1{\alpha}$ production significantly in comparison to DNP-IgE control group in a dose dependent manner. IL-13 production was inhibited at a concentration of 200 ${\mu}g/ml$, 400 ${\mu}g/ml$ and MIP-$1{\alpha}$ was inhibited at a concentration of 100 ${\mu}g/ml$, 200 ${\mu}g/ml$, 400 ${\mu}g/ml$. Western blot analysis of transcription factors involving Th2 cytokines expression revealed prominent decrease of the mast cell specific transcription factors including NFAT-1, c-Jun as well as NF-${\kappa}B$ p65 but not NFAT-2 and c-Fos. Conclusion These results indicate that PRAL has the effect of suppressing Th2 cytokines production in the MC/9 mast cells. These data represent that PRAL potentiates therapeutic activities to the allergic disease by regulating Th2 cytokines in the MC/9 mast cells.

• 한국식품영양과학회지
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• 제34권8호
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• pp.1188-1194
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• 2005

Streptozotocin으로 당뇨를 유발시킨 생쥐에서 fructan(치커리이뉼린과 치커리이뉼린올리고당, 프럭토올리고당)이 혈당, 인슐린과 소장 및 신장의 이당류분해효소 활성 및 면역능에 미 치는 영향을 조사한 결과는 다음과 같이 요약할 수 있다. 혈당은 당뇨군들이 정상군보다 유의하게 높았으며, 치커리이뉼린군의 혈당이 당뇨대조군에 비하여 유의하게 낮았다. Fructan은 혈장 인슐린 농도에는 영향을 미치지 않았다. 소장점막의 maltase와 sucrase활성은 당뇨군들이 정상군에 비하여 유의적으로 높았으나, 당뇨군 중에서 치커리이뉼린군이 가장 낮은 경향을 보였다. 신장 maltase 활성은 정상군과 당뇨군이 유사한 수준이었으나, 신장 sucrase와 lactase 활성은 정상군에 비하여 당뇨군에서 유의하게 높은 수준을 보였으며, 치커리이뉼린군이 당뇨대조군에 비하여 유의하게 낮은 수준을 보였다. LPS에 의한 비장세포증식에서는 당뇨대조군에 비하여 치커리이뉼린올리고당군이 유의 적으로 증가하여 정상군과 비슷한 수준을 보였다. ConA에의한 비장세포증식은 군간 유의적인 차이를 보이지 않았으나 LPS에 의한 증식 결과와 비슷한 경향을 보였다. 비장세포로부터의 IL-2의 분비는 당뇨군들이 정상군에 비하여 낮았으나, 당뇨군간에는 유의한 차이가 없었다. 결론적으로 당뇨 생쥐에서 프럭토올리고당, 치커리이뉼린올리고당, 치커리이뉼린을 비교한 결과, 혈당 강하에 미치는 효과가 치커리이뉼린에서 유의하게 나타났으며, 비장세포 증식능은 정상군에 비하여 당뇨군에서 감소하는 경향을 보였으나, 치커리이뉼린올리고당 섭취군이 당뇨대조군에 비해 비장세포 증식능이 유의하게 증가하여 면역능이 개선되는 효과를 보였다.

• International Journal of Oral Biology
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• 제34권1호
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• pp.1-6
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• 2009

The purpose of the present study was to examine the role of peripheral nitric oxide (NO) pathways in the onset of interleukin (IL)-1$\beta$-induced mechanical allodynia in the orofacial area. Experiments were carried out on male Sprague-Dawley rats weighing 230-280 gm and surgical procedures were performed under pentobarbital sodium (40 mg/kg, i.p.). Under anesthesia, a polyethylene tube (PE10) was implanted into the subcutaneous area of one vibrissa pad, which enabled the injection of IL-1$\beta$ or other chemicals. We subcutaneously injected 50 ${\mu}L$ of IL-1$\beta$ into a vibrissa pad through the implanted polyethylene tube with a 100 ${\mu}L$ Hamilton syringe. After the administration of 0.01, 0.1, 1, or 10 pg of IL-1$\beta$, withdrawal behavioral responses were examined. The subcutaneous injection of saline had no effects on the air-puff thresholds. Following the subcutaneous injection of 0.01, 0.1, 1, or 10 pg of IL-1$\beta$, the threshold of air puffs decreased significantly to 12 $\pm$ 3, 7 $\pm$ 2, 5 $\pm$ 1, or 5 $\pm$ 1 psi, respectively, in a dose dependent manner. Pretreatment with L-NAME, a nitric oxide synthase (NOS) inhibitor, blocked IL-1$\beta$-induced mechanical allodynia. However, neither D-NAME, an inactive isomer of L-NAME, nor vehicle affected the IL-1$\beta$-induced mechanical allodynia. Subcutaneous injection of IL-1$\beta$ increased the number of c-fos-like immunoreactive neurons, whereas pretreatment with L-NAME decreased this number, in the trigeminal caudal nucleus. These results suggest that pro-inflammatory cytokines and NO are important contributors to the pathogenesis of persistent and exaggerated IL-1$\beta$-induced pain states. Based on these observations, peripheral application of NOS inhibitors may be of therapeutic value in treating pain disorders in the clinic.

• 한방재활의학과학회지
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• 제25권1호
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• pp.27-44
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• 2015

Objectives This study was carried out to find the effects of Gami-cheongyulsaseub-tang (hereinafter referred to GCST) on the inhibition of zymosan-induced pain in rats and collagen II-induced arthritis (CIA) in DBA/1J mouse. Methods As an acute inflammatory pain model, peripheral inflammation was induced by intraplantar injection of zymosan into the right hind paw in rats and then the hyperalgesia and pain regulating factors in spinal cord were analyzed. As a chronic inflammation model, the mixture of collagen II and complete Freund's adjuvant was treated into mice to establish rheumatoid arthritis and then body weight, thickness of hind paw, pathological change of spleen, immunological rheumatoid factor (IgG1, IgG2a, IgG2b, IgM and anti-collagen II), pro-inflammatory cytokines, and bone injury were analyzed. Results In the acute inflammatory pain model, GCST significantly inhibited the thermal and mechanical hyperalgesia and the pain regulating factors, including Fos, CD11b, PKA and PKC, in the spinal cord with a dose-dependent manner. In the chronic rheumatoid arthritis model, GCST administration decreased arthritic index and paw edema as compared with CIA control group. In particular, GCST reduced significantly the serum levels of total IgG2a, IgG2b, IgM, and specific anti-collagen II, but not total IgG1. GCST also resulted in the attenuation of bone injury and spleen enlargement/adhesion in CIA mice. Moreover, the secretion of pro-inflammatory cytokines TNF-${\alpha}$ and IL-$1{\beta}$ in CIA mice was significantly reduced by GCST in a dose-dependent manner. Conclusions Comparison of the results in this study showed that GCST had anti-nociceptive and immunomodulatory effects. These data imply that GCST can be used as an effective drug for not only rheumatoid arthritic pain but also other auto-immune diseases.

• Journal of Acupuncture Research
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• 제17권2호
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• pp.169-186
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• 2000

To study anti-cancer effect and molecular biological mechanism of bee venom for aqua-acupuncture, the effects of bee venom on cell viability, apoptosis, and cell cycle were analyzed using MTT assay, tryphan blue assay, [3H]thymidine release assay, flow cytometric analysis, activity of caspase-3 protease activity assay, and immunocytometric analysis of PCNA. To explore whether anti-cancer effects of bee venom are associated with the transcriptional control of gene expression, quantitative RT-PCR analysis of apoptosis- and cell cycle-related genes was performed. The obtained results are summarized as follows: 1. The MTT assay demonstrated that cell viability was decreased by bee venom in a dose-dependant manner. 2. Significant induction of apoptosis was identified using tryphan blue assay, [$^3H$]thymidine release assay, and flow cytometric analysis of sub $G_1$ fraction. 3. In analysis of caspase-3 protease activity, the activity had increased significantly, in a dose-dependant manner. 4. Quantitative RT-PCR analysis of the apoptosis-related genes showed that Bcl-2 and $Bcl-X_L$ were down-regulated whereas Bax was up-regulated by bee venom treatment. 5. In flow cytometric analysis of cell cycle and immunocytometric analysis of PCNA expression, cell numbers of $G_1$ phase was increased by a dose-dependant manner. 6. In quantitative RT-PCR analysis of the cell cycle-related genes, p21, p27, and p57 were increased, while Cyclin D1, CDK4, c-Myc, c-Fos, and Histone H3 were decreased. In contrast, there were no remarkable changes in expression levels of CDC2 and c-Jun.

• Clinical and Experimental Reproductive Medicine
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• 제30권4호
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• pp.309-316
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• 2003

연구목적: 스테로이드 계통의 에스트로겐 호르몬은 막 수용체와 결합하고 DNA에 부착되어, 자궁조직에서 발현되는 많은 유전자들의 발현 양상을 조절하는 것으로 알려져 있다. 본 연구에서는 난소를 제거한 생쥐 모델을 이용하여 에스트로겐에 의해 조절되는 전사 관련 유전자(transcription factor)들을 동정하고, early up-regulation gene으로 확인된 Fos related antigen (Fra1과 Fra2) 유전자의 발현 양상을 RT-PCR과 면역염색방법으로 살펴보았다. 연구재료 및 방법: 난소 절제술을 시행한 생쥐에 에스트로겐을 피하주사하고 2, 4, 6, 12시간 간격으로 자궁조직을 적출하였다. 대조군으로는 sesame oil만을 주사한 후 2시간째에 수획한 자궁조직을 사용하였으며, 시간대별로 채취한 자궁조직(n=4)에서 RT-PCR을 수행하였다. RT-PCR을 통해 early response gene으로 확인된 Fra1과 Fra2에 대한 에스트로겐의 영향을 살펴보기 위해 estrogen receptor antagonist인 ICI 182, 780을 주사하여 유전자 발현 양상의 변화를 살펴보았다. 또한, 자궁조직내에서의 단백질 발현 부위를 관찰하기 위해 면역조직화학염색을 실시하였다. 결 과: 생쥐 자궁조직에서 에스트로겐에 의해 발현 양상의 변화가 확인된 유전자는 early up-regulation genes (CREB2, Fra-1, 2, GATA5), late up-regulation gene (E2F1), no response genes (CREB1, ATF1, GLI3, E2F3), down-regulation genes (GLI2, E2F5, GATA-2, 3, 6) 등으로 구분할 수 있었다. 그 중 early up-regulation genes에 해당하는 Fra1과 Fra2 유전자는 ICI 182, 780에 의해 그 발현이 유의하게 감소되는 것을 확인하였다(p < 0.01). 이들 단백질은 생쥐 자궁조직의 상피세포층, 기질층, 근육층에서 고루 발현되었으며, 특히 근육층에서 강한 염색정도를 관찰할 수 있었다. 결 론: 이상의 결과를 통해 Fra1과 Fra2 유전자의 발현은 에스트로겐에 의해 조절됨을 알 수 있었으며, 이들의 강한 발현이 자궁조직의 근육층에서 관찰되어 이들의 기능에 대한 연구가 필요할 것으로 생각된다.

• 한국식품과학회지
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• 제32권2호
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• pp.277-283
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• 2000

콩나물의 성장은 콩의 수침 및 주수시 용존오존농도 0.3 ppm으로 처리하였을때가 가장 양호하였다. 콩의 수침시 용존오존농도 0.3 ppm으로 30분간 처리하여 5일간 재배한 콩나물의 배축신장도는 무처리보다 19%가 높았으며 시루당의 총중량은 27%가 증가하였다. 반면에 뿌리의 중량은 27%가 감소하였다. 그러나, 수침시와 주수시 처리와 수침시 처리 사이의 성장도 차이는 뚜렷하지 않았다. Carbendazim, captan, diazinon, fenthion, dichlorvos 및 chlorpyrifos 등 6종의 농약을 $4.75{\sim}8.35\;ppm$으로 오염시킨 콩의 수침과정 및 5일간 재배중의 잔류농약의 자연분해율은 $85{\sim}99%$였으나 주수시 0.3 ppm의 오존수로 처리하였을 때는 $89{\sim}100%$, 수침시와 주수시에 처리하였을 때는 $94{\sim}100%$이였다. 또 농약별 오존수에 의한 분해율은 captan>dichlorvos>fenthion>carbendazim>diazinon>chlorpyrifos순이였다.

• 한국전통의학지
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• 제15권1호
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• pp.113-127
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• 2006

Objectives: This study was produced to examine the effects of moxibustion that had been played important role to traditional oriental medical treatment on disease. Recently, it was reported that moxi-tar which is generated in the process of moxibustion as burning combustibles decreased nitric oxide(NO) and inducible NO synthase (iNOS) generation in cellular experiments. Methods: Carrageenan-induced arthritis rat model was used to test the effect of moxi-tar as a chronic pain model. Diluted moxi-tar was single injected in several acupoints or combined with electroacupuncture (l ms, 2 Hz, and 2 mA) into contralateral ST36 acupoint for 30 min to assess the synergic effects. After the treatment, behavioral tests measuring stepping force were periodically conducted during the next 12 hours. Endogenous NO and iNOS, cyclooxygenase-2 (COX-2), and c-Fos protein expression in the spinal cord were examined on a rat model of carrageenan-induced arthritis. Results : After the induction of arthritis, rats subsequently showed a reduced stepping force of the affected limb for at least the next 4 days. The reduced stepping force of the limb was presumably due to a painful knee, since oral injection of indomethacin produced temporary improvement of weight bearing. Maxi-tar produced significant improvement of stepping force of the hindlimb affected by the arthritis lasting at least 9 hours. The magnitude of this improvement was equivalent to that obtained after an oral injection of 3 mg/kg of indomethacin and this improvement of stepping force was interpreted as an analgesic effect. Maxi-tar produced the improvement of stepping force of the affected hindlimb in a dose-dependent manner. Both NO production and iNOS, COX-2 protein expression increased by arthritis were suppressed by maxi-tar. Moxi-tar on combination with electroacupuncture (EA) produced more powerful and longer lasting improvement of stepping force of the hindlimb affected by the arthritis than either moxi-tar or EA did. Conclusion : The present study suggest that maxi-tar produces a potent analgesic effect on the chronic knee arthritis pain model in the rat and that moxi-tar-induced analgesia modulate endogenous NO through the suppression of iNOS/COX-2 protein expression.

• 대한임상검사과학회지
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• 제51권3호
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• pp.323-328
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• 2019

결핵균에 의한 만성 세균성 감염인 결핵은 결핵균의 특성, 숙주의 면역상태와 유전적 감수성의 차이에 의해 잠복성과 활동성으로의 진행정도에 차이가 있다. 결핵균에 대한 숙주 방어 기전은 주로 대식세포, T 세포 및 수지상 세포 사이의 상호 작용에 기인한다. CD44는 결핵균에 감염되면 활성 T 세포에서 발현되며 림프구 이동을 조절한다. 또한 CD44는 ECM에 대한 백혈구의 부착을 매개하여 대식세포, CD4+ T cell 등을 폐로 불러모으는 역할을 한다. 따라서, CD44 유전자의 다형성은 결핵균에 대한 숙주세포의 면역기전 저하를 유발할 수 있다. 이 연구의 목적은 CD44 유전자의 유전자 다형성이 결핵의 감수성에 영향을 미치는지 조사하는 것이다. 결핵균과 CD44의 연관성에 대하여 대한 한국 협회 자원의 443명의 cases와 3228명의 control을 이용하여 CD44 유전자의 237개의 SNP를 분석하였다. 이 중 17개의 SNP가 결핵과 통계적으로 유의한 관련성을 보였다. 가장 유의성 있는 SNP는 rs75137824였다(OR=0.231, CI: 1.51~3.56, $P=1.3{\times}10^{-4}$). 또한 결핵 발병에 유의성이 있는 SNP중 rs10488809의 경우는 전사인자 JUND 및 FOS에 결합하는 부위로써 CD44 유전자 발현에 영향을 줄 수 있는 것으로 확인할 수 있었다. 이러한 결과는 결핵 발병이 CD44 발현 차이에 의한 숙주 면역반응에 차이에 의해서 감수성의 차이가 있을 수 있음을 나타 낼 수 있다. 이번 연구 결과는 결핵균 감염에 대한 숙주면역의 유전적 차이가 결핵 진행정도의 차이를 유발할 수 있다는 유전적 배경에 대한 기반을 마련해 줄 수 있을 것이라고 기대한다.

• Animal Bioscience
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• 제35권7호
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• pp.964-974
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• 2022

Objective: The highly pathogenic avian influenza virus (HPAIV) is a threat to the poultry industry and economy and remains a potential source of pandemic infection in humans. Antiviral genes are considered a potential factor for studies on HPAIV resistance. Therefore, in this study, we investigated gene expression related to the mitogen-activated protein kinase (MAPK) signaling pathway by comparing non-infected, HPAI-infected resistant, and susceptible Ri chicken lines. Methods: Resistant (Mx/A; BF2/B21) and susceptible Ri chickens (Mx/G; BF2/B13) were selected by genotyping the Mx and BF2 genes. Then, the tracheal tissues of non-infected and HPAIV H5N1 infected chickens were collected for RNA sequencing. Results: A gene set overlapping test between the analyzed differentially expressed genes (DEGs) and functionally categorized genes was performed, including biological processes of the gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathways. A total of 1,794 DEGs were observed between control and H5N1-infected resistant Ri chickens, 432 DEGs between control and infected susceptible Ri chickens, and 1,202 DEGs between infected susceptible and infected resistant Ri chickens. The expression levels of MAPK signaling pathway-related genes (including MyD88, NF-κB, AP-1, c-fos, Jun, JunD, MAX, c-Myc), cytokines (IL-1β, IL-6, IL-8), type I interferons (IFN-α, IFN-β), and IFN-stimulated genes (Mx1, CCL19, OASL, and PRK) were higher in H5N1-infected than in non-infected resistant Ri chickens. MyD88, Jun, JunD, MAX, cytokines, chemokines, IFNs, and IFN-stimulated expressed genes were higher in resistant-infected than in susceptible-infected Ri chickens. Conclusion: Resistant Ri chickens showed higher antiviral activity compared to susceptible Ri chickens, and H5N1-infected resistant Ri chickens had immune responses and antiviral activity (cytokines, chemokines, interferons, and IFN-stimulated genes), which may have been induced through the MAPK signaling pathway in response to H5N1 infection.