• Journal of Microbiology and Biotechnology
• /
• 제19권11호
• /
• pp.1401-1407
• /
• 2009

Many Lactobacillus strains have been promoted as good probiotics for the prevention and treatment of diseases. We engineered recombinant Lactobacillus casei, producing biologically active canine granulocyte macrophage colony stimulating factor (cGM-CSF), and investigated its possibility as a good probiotic agent for dogs. Expression of the cGM-CSF protein in the recombinant Lactobacillus was confirmed by SDS-PAGE and Western blotting methods. For the in vivo study, 18 Beagle puppies of 7 weeks of age were divided into three groups; the control group was fed only on a regular diet and the two treatment groups were fed on a diet supplemented with either $1\times10^9$ colony forming units (CFU)/day of L. casei or L. casei expressing cGM-CSF protein for 7 weeks. Body weight was measured, and fecal and blood samples were collected from the dogs during the experiment for the measurement of hematology, fecal immunoglobulin (Ig)A and IgG, circulating IgA and IgG, and canine corona virus (CCV)-specific IgG. There were no differences in body weights among the groups, but monocyte counts in hematology and serum IgA were higher in the group receiving L. casei expressing cGM-CSF than in the other two groups. After the administration of CCV vaccine, CCV-specific IgG in serum increased more in the group supplemented with L. casei expressing cGM-CSF than the other two groups. This study shows that a dietary L. casei expressing cGM-CSF enhances specific immune functions at both the mucosal and systemic levels in puppies.

• 한국산학기술학회논문지
• /
• 제16권1호
• /
• pp.365-370
• /
• 2015

GM-CSF는 중요한 조혈모세포 성장인자로서 면역요법에서 중요한 기능을 한다. 본 연구의 목적은 GM-CSF가 돼지 처녀생식배아의 발달과 세포 수 및 착상관련 유전자의 발현에 관한 영향을 평가하는 것이다. 본 연구에서 돼지 처녀 활성화 배아는 GM-CSF가 5, 10, 20 ng/ml 존재 하에서 7일 동안 배양하여 배 반포의 형성율과 전 세포 수 그리고 유전자 발현을 평가하였다. 그 결과 단백질이 없는 배양액에 20 ng/ml의 GM-CSF를 첨가 하였을 때 배 반포의 형성 율이 유의적으로 증가하였으며 배반포의 세포 수 또한 GM-CSF 를 첨가한 배양액에서 증가 하였다. GM-CSF는 처녀생식 배 반포에서 interleukin-6의 mRNA 발현을 증가 시켰으나, LIF 수용체 mRNA 발현에는 영향을 주지 않는다는 것을 real time RT-PCR로 밝혀내었다. 이 결과로 GM-CSF 성분이 확인된 배양액에서 돼지 배아의 체외 발달 과 생존력을 강화 시켰음을 시사하고 있다.

• 동의생리병리학회지
• /
• 제28권1호
• /
• pp.29-34
• /
• 2014

Bone homeostasis is maintained by balance between bone resorbing-osteoclasts and bone forming-osteoblasts. Excessive osteoclastic bone resorption plays a critical role in bone destruction in pathological bone diseases such as osteoporosis, rheumatoid arthritis, and periodontal disease. Many compounds derived from natural products have pharmacological applications and have therapeutic value for treating or preventing several bone diseases characterized by excessive bone resorption. To discover new compounds that can act as anti-resorptive agents, we screened for natural compounds that regulate osteclast differentiation, and found that water extract of Ziziphus Jujuba Mill (WEZJ) has inhibitory effects on osteoclast differentiation. In this study, WEZJ clearly inhibits the osteoclast differentiation in the presence of receptor activator of nuclear factor kB (RANKL), macrophage colony-stimulating factor (M-CSF) without cytoxicity by blocking activation of nuclear factor of activated T cells (NFAT)c1, and c-Fos. In signaling pathway, the phosphorylation of Akt, p38, c-Jun N-terminal kinases (JNK), extracellular signal-regulated kinases (ERK) and the expression of osteoclast-associated receptor (OSCAR), tartrate-resistant acid phosphates (TRAP), Integrin av, Integrin b3, Cathepsin K are suppressed, too. These result suggest that WEZJ may have therapeutic value for treating or preventing several bone diseases characterized by excessive bone destruction.

• 한방안이비인후피부과학회지
• /
• 제25권3호
• /
• pp.1-12
• /
• 2012

Objectives : Fagopyrum esculentum(FE) has been used for removal of inflammation of internal organs and treatment of sore and ulcer by heat toxin in Korean herbal medicines. In this study, To investigated the protective effect of FE on allergic response, we determined whether FE inhibits allergic response. Methods : The effect of FE was analyzed by ELISA, RT-PCR and Western blot in RBL-2H3 cells. We investigated cell viability, ${\beta}$-hexosaminidase, as a marker of degranulation, cytokne, and intracellular ROS and MAPK and NF-${\kappa}B$ signaling. Results : We found that FE suppressed ${\beta}$-hexosaminidase release, the production of IL-4 and TNF-${\alpha}$ and intracellular ROS level in RBL-2H3 by the anti-DNP IgE plus DNP-HSA stimulation. FE also significantly inhibited cytokine mRNA expressions, such as IL-$1{\beta}$, IL-2, IL-3, IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$ and GM-CSF in RBL-2H3. In addition, PF suppressed the phospholyation of ERK1/2, JNK1/2, p38 and $I{\kappa}B{\alpha}$ and NF-${\kappa}B$ signal transduction pathway. Conclusions : Our results indicate that FE protects against allergic response and exerts an anti-inflammatory effect through the inhibition of degranulation and production of cytokines and ROS via the suppression MAPK and NF-${\kappa}B$ of signal transduction. Abbrevations : FE, Fagopyrum esculentum; RBL-2H3, rat basophilic leukemia cell line; ROS, reactive oxygen species; MAPK, Mitogen-activated protein kinase; $NF{\kappa}B$, nuclear factor ${\kappa}B$; $TNF{\alpha}$, Tumor necrosis factor alpha; GM-CSF, Granulocyte macrophage colony-stimulating factor; ERK, extracellular-signal-regulated kinase; JNK, c-Jun NH2-terminal kinase; p38, p38 MAP kinase; $I{\kappa}B{\alpha}$, inhibitory-kappa B alpha.

• Applied Microscopy
• /
• 제39권3호
• /
• pp.253-260
• /
• 2009

상처치유는 growth factor와 cytokine들의 복합적인 작용에 의해 일어나는 과정이다. GM-CSF는 호중구와 대식세포를 분화시키고 각질세포를 증식시키는 다면발현성 사이토카인으로 여러 세포에서 재조합되어 사용되고 있다. (주)핸슨바이오텍에서 개발한 rice cell 유래의 재조합 rrhGM-CSF는 다른 세포에서 유래된 GM-CSF들보다 탄수화물 측쇄가 많은 것이 특징이다. rrhGM-CSF의 창상치유효과를 알아보기 위하여 햄스터에 만성 당뇨를 유도하여 상처치유가 어려운 동물모델을 만든 후, 등쪽에 인위적으로 피부 전 층을 제거하여 창상을 유발하였다. 실험군에는 rrhGM-CSF가 포함된 연고를 적용하고, 대조군에는 base cream만을 적용하여 재상피화되는 과정과 피부조직의 구조 및 PCNA 반응을 통한 세포증식 정도를 비교하였다. 육안 및 조직학적 관찰 결과, rrhGM-CSF를 적용한 그룹의 재상피화 속도가 더 빨랐으며, PCNA 면역조직화학적 조사에서도 증식하는 각질세포의 수가 대조군보다 상대적으로 많은 것으로 나타남으로써, rrhGM-CSF에 의한 피부상처 치유 효과를 관찰할 수 있었다. 이러한 결과들로 미루어 보아 벼세포 유래의 rrhGM-CSF는 국소적인 피부 창상 치유에 효과적인 것으로 보이며, rrhGM-CSF의 풍부한 당화 사슬은 이 단백질이 상처부위에 안정된 상태로 기능을 나타냄으로써 효과를 오랫동안 지속시킬 수 있었던 것으로 사료된다.

• 대한본초학회지
• /
• 제37권3호
• /
• pp.9-19
• /
• 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.

• 생명과학회지
• /
• 제30권3호
• /
• pp.267-277
• /
• 2020

이 연구는 설치류에서 양파추출물의 효능을 평가하기 위하여 실시하였다. 실험동물은 대조군(control)과 양파추출물 투여군(ACE)으로 나누어서 진행하였다. ACE그룹은 대조군에 비해 적혈구(RBC), 헤모글로빈(HGB), 헤마토크릿(Hct) 수준이 약간 증가하는 것을 보였다(p<0.05). 반면, 헤모글로빈, 단핵구, 림프구 및 호중구는 대조군과 비교하였을 경우 유의미한 변화(p<0.05)가 없었다. GOT (glutamic oxaloacetic transaminase) 및 GPT (glutamic pyruvic transaminase) 수준을 분석한 결과 대조군에 비해 ACE그룹이 유의적으로 감소하였다(p<0.05). 혈당, 총단백질, HDL-콜레스테롤은 ACE 그룹에서 약간 높았고, 트리글리세라이드, 총콜레스테롤 수치는 대조군에 비해 ACE그룹에서 더 낮았다(p<0.05). 간 조직과 혈액의 면역과 염증에 관련된 사이토카인(인터루킨 1알파(IL-1α), 인터루킨 1베타(IL-1β), 종양괴사인자알파(TNF-α), 인터루킨 6(IL-6), 인터루킨 10(IL-10), 인터페론감마(INF-γ), 인터루킨 18(IL-18), 인터루킨 2(IL-2), 과립구 대식구 집락자극인자(GM-CSF)) 수준은 모두 정상 범위 내에 있는 것으로 확인되었다. 본 연구결과는 고농축 식이성 양파추출물은 혈액학적 지표와 면역 기능에 독성이 없다는 것을 보여주는 기초데이터로 활용할 수 있을 것이다.

• Maxillofacial Plastic and Reconstructive Surgery
• /
• 제38권
• /
• pp.48.1-48.8
• /
• 2016

Background: This study investigates the effect of alendronate-treated osteoblasts, as well as the effect of low-level laser therapy (LLLT) on the alendronate-treated osteoblasts. Bisphosphonate decreases the osteoblastic activity. Various treatment modalities are used to enhance the bisphosphonate-treated osteoblasts; however, there were no cell culture studies conducted using a low-level laser. Methods: Human fetal osteoblastic (hFOB 1.19) cells were treated with $50{\mu}M$ alendronate. Then, they were irradiated with a $1.2J/cm^2$ low-level Ga-Al-As laser (${\lambda}=808{\pm}3nm$, 80 mW, and 80 mA; spot size, $1 cm^2$; NDLux, Seoul, Korea). The cell survivability was measured with the MTT assay. The three cytokines of osteoblasts, receptor activator of nuclear factor ${\kappa}B$ ligand (RANKL), osteoprotegerin (OPG), and macrophage colony-stimulating factor (M-CSF) were analyzed. Results: In the cells treated with alendronate at concentrations of $50{\mu}M$ and higher, cell survivability significantly decreased after 48 h (p < 0.05). After the applications of low-level laser on alendronate-treated cells, cell survivability significantly increased at 72 h (p < 0.05). The expressions of OPG, RANKL, and M-CSF have decreased via the alendronate. The RANKL and M-CSF expressions have increased, but the OPG was not significantly affected by the LLLT. Conclusions: The LLLT does not affect the OPG expression in the hFOB cell line, but it may increase the RANKL and M-CSF expressions, thereby resulting in positive effects on osteoclastogenesis and bone remodeling.

• 대한한방내과학회지
• /
• 제23권1호
• /
• pp.15-23
• /
• 2002

Objective : We aimed to identify the dose-dependent inhibitory effects of Sabaek-san(瀉白散) and Sabaeksan plus Sasam(Adenophorae Radix) 瀉白散加沙蔘) on the mRNA expressions of Interleukin(IL)-6, IL-8 and granulocyte macrophage colony stimulating factor(GM-CSF) involved in the asthma model. Materials and Methods : Through this study, BEAS-2B cell lines, human epithelial cells were used. These cells were stimulated by tumor necrosis factor(TNF)-${\alpha}$, IL-1${\beta}$ and histamine for artificial inflammatory expression. ${\beta}$-action messenger RNA(mRNA) was used for standards. After each 24hours of Sabaeksan and Sabaeksan plus Sasam treatment, total cellular RNAs were collected by treating RNA zol directly on living cells, Then the transcriptional activities of IL-6, 8 and GM-CSF were measured by RT-PCR with electrophoresis, Results : The mRNA expressions of IL-6 are significantly inhibited compared to those of controlled group at 40 and 100ug/ml of Sabaeksan extract and $100{\mu}g/ml$ of Sabaeksan plus Sasam extract (p<0.05). The mRNA expressions of IL-8 are significantly inhibited compared to that of controlled group at 2.40 and 100 ug/ml of Sabaeksan extract and $40.100{\mu}g/ml$ of Sabaeksan plus Sasam extract(p<0.05) THe mRNA expressions of GM-CSF are significantly inhibited compared to those of the controlled group at $100{\mu}g/ml$ of Sabaeksan extract adn $40.100{\mu}g/ml$ of Sabaeksan plus Sasam extract.(p<0.05) Conclusions : This study shows that Sabaeksan and Sabaeksan plus Sasam have dose-dependent inhibitory effects on the mRNA expressions of IL-6, IL-8 and GM-CSF in human epithelial cells. Therefore, these types of herb medicine may inhibit the inflammatory process of asthma. Advanced studies are required to investigate the mechanisms of inhibition by herb medicine in the asthma model.

• 대한한방내과학회지
• /
• 제22권3호
• /
• pp.415-422
• /
• 2001

Objectives: We aimed to identify the dose-dependent inhibitory effects of Yukmijihwang-tang-Hap-Sabaek-san(YMHSB) and Root cortex of Morus alba L.(RCM) on the mRNA expression of Interieukin(IL)-6, IL-S, granulocyte macrophage colony stimulating factor(GM-CSF) involved in the asthma model. Methods: In this study BEAS-2B cell lines, human epithelial cells, were used. These cells were stimulated by tumor necrosis $factor(TNF)-{\alpha},\;IL-1{\beta}$ and histamine for artificial inflammatory expression. ${\beta}-actin$ messenger RNA(mRNA) was used for the internal standard. After each 24 hours of the YMHSB and RCM treatment, total cellular RNAs were collected by treating RNA zol directly on the living cells. Then the transcriptional activities of IL-6, 8 and GM-CSF were measured by RT-PCR with electrophoresis. Results: In the YMHSB study, the mRNA expression of GM-CSF and IL-8 is significantly inhibited compared to that of control group. But the mRNA expression of IL-6 is not significantly inhibited. In the RCM study, the mRNA expression of GM-CSF and IL-S is significantly inhibited compared to that of control group. But the mRNA expression of IL-6 is not significantly inhibited. Conclusions: This study shows that YMHSB and RCM have dose-dependent inhibitory effects on the mRNA expression of IL-S and GMCSF in human epithelial cells. So these herbal medicines may inhibit the inflammatory process of asthma. Advanced studies are required to investigate the mechanisms of inhibition by herbal medicine in the asthma model.