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Inhibitory effects of Broussonetia kazinoki twig extract on allergic inflammatory reactions in TNF-𝛼/IFN-𝛾-stimulated HaCaT and IgE-sensitized RBL-2H3 cells (TNF-𝛼/IFN-𝛾로 자극된 HaCaT 및 IgE로 감작된 RBL-2H3 세포에서 닥나무 가지 추출물의 알러지 염증반응 억제 효과)

  • Won-Bin Bae;Eun-Hye Kim;Min-Ju Kim;Seun-Ah Yang
    • Food Science and Preservation
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    • v.31 no.2
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    • pp.307-314
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    • 2024
  • Broussonetia kazinoki twig extract (BKT) is recognized for its antioxidant and anti-cancer effects and natural whitening properties. So, it is used as a raw material for cosmetics. B. kazinoki twig is also an edible raw material. B. kazinoki has been used in Asia for paper production and oriental medicine, has anti-diabetic effects, and contains various flavonoids and alkaloids. In this study, to evaluate the efficacy of BKT on allergic skin inflammatory responses, we investigated its effects on factors related to skin inflammation in HaCaT keratinocytes and allergic responses in RBL-2H3 cells. There was no cytotoxicity of the 70% ethanol extract against HaCaT and RBL-2H3 cells. In HaCaT cells, stimulation with tumor necrosis factor-alpha (TNF-𝛼) and interferon-gamma (IFN-𝛾) increased the production of several chemokines, including thymus and activation-regulated chemokine (TARC), macrophage-derived chemokine (MDC), and regulated on activation, normal T cell expressed and secreted (RANTES). However, it was observed that this elevation was notably mitigated in a concentration-dependent manner upon treatment with BKT. Furthermore, BKT treatment demonstrated a significant reduction of 𝛽-hexosaminidase and inflammatory cytokines TNF-𝛼 and IL-4 in IgE-sensitized RBL-2H3 cells. Thus, it is expected that BKT can be used as a natural cosmetic and food ingredient that effectively suppresses allergic inflammatory reactions.

Intranasal and intraperitoneal immunization against Brucella infection using niosome and mannosylated niosomes containing Brucella recombinant trigger factor/Bp26/Omp31 chimeric protein in a mouse model

  • Fahimeh Sharif;Razieh Nazari;Mahdi Fasihi-Ramandi;Ramezan Ali Taheri;Mohsen Zargar
    • Clinical and Experimental Vaccine Research
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    • v.13 no.3
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    • pp.232-241
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    • 2024
  • Purpose: Brucellosis, a zoonotic infectious disease, is a worldwide health issue affecting animals and humans. No effective human vaccine and the complications caused by the use of animal vaccines are among the factors that have prevented the eradication of the disease worldwide. However, bio-engineering technologies have paved the way for designing new targeted and highly efficacious vaccines. In this regard, the study aimed to evaluate immunity induced by mannosylated niosome containing Brucella recombinant trigger factor/Bp26/Omp31 (rTBO) chimeric protein in a mouse model. Materials and Methods: rTBO as chimeric antigen (Ag) was expressed in Escherichia coli BL21 (DE3) and, after purification, loaded on niosome and mannosylated niosome. The characteristics of the nanoparticles were assessed. The mice were immunized using rTBO, niosome, and mannosylated niosome-rTBO in intranasal and intraperitoneal routes. Serum antibodies (immunoglobulin [Ig]A, IgG, IgG1, and IgG2a) and splenocyte cytokines (interferon-gamma, interleukin [IL]-4, and IL-12) were evaluated in immunized mice. Finally, immunized mice were challenged by B. melitensis and B. abortus. A high antibody level was produced by niosomal antigen (Nio-Ag) and mannosylated noisomal antigen (Nio-Man-Ag) compared to the control after 10, 24, and 38 days of immunization. The IgG2a/IgG1 titer ratio for Nio-Man-Ag was 1.2 and 1.1 in intraperitoneal and intranasal methods and lower than one in free Ag and Nio-Ag. Cytokine production was significantly higher in the immunized animal with Ag-loaded nanoparticles than in the negative control group (p<0.05). Moreover, cytokine and antibody levels were significantly higher in the injection than in the inhalation method (p<0.05). Results: The combination of mannosylated noisome and rTBO chimeric proteins stimulate the cellular and humoral immune response and produce cytokines, playing a role in developing the protective acquired immune response in the Brucella infectious model. Also, the intraperitoneal route resulted in a successful enhancement of cytokines production more than intranasal administration. Conclusion: Designing an effective vaccine candidate against Brucella that selectively induces cellular and humoral immune response can be done by selecting a suitable nanoniosome formulation as an immunoadjuvant and recombinant protein as an immune response-stimulating Ag.

Lymphocyte Proportion and Cytokines from the Bone Marrow of Patients with Far-Advanced Pulmonary Tuberculosis with Peripheral Lymphocytopenia (말초혈액의 림프구감소증을 동반한 중증폐결핵 환자들에서 골수 내의 림프구 분획과 사이토카인 소견)

  • An, Chang Hyeok;Kyung, Sun Yong;Lim, Young Hee;Park, Gye Young;Park, Jung Woong;Jeong, Sung Hwan;Ahn, Jeong Yeal
    • Tuberculosis and Respiratory Diseases
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    • v.55 no.5
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    • pp.449-458
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    • 2003
  • Background : The poor prognostic factors of far-advanced pulmonary tuberculosis(FAPTB) are lymphocytopenia in the peripheral blood(PB)(< $1,000/mm^3$) and $T_4$-cell count ${\leq}500/mm^3$. However, the cause of PB lymphocytopenia in FAPTB is unclear. The aim of this study was to analyze the lymphocyte proportion and cytokines of the bone marrow(BM) in FAPTB patients with peripheral lymphocytopenia in order to clarify whether the limiting step of the lymphocytopenia occurs in production, differentiation, or circulation. Methods : This study included patients with FAPTB between August 1999 and August 2002 who visited Gachon Medical School Gil Medical Center. The exclusion criteria were old age(${\geq}65years$), cachexia or a low body weight, shock, hematologic diseases, or BM involvement of tuberculosis. The distributions of cells in PB and BM were analyzed and compared to the control group. The interleukin(IL)-2, IL-7, IL-10, TNF-${\alpha}$, Interferon-${\gamma}$, and TGF-${\beta}$ levels in the BM were measured by ELISA. Result : Thirteen patients(male : female=9:4) were included and the mean age was $42{\pm}12$years. The proportion and count of the lymphocytes in the PB were significantly lower in the FAPTB group ($7.4{\pm}3.0%$, $694{\pm}255/mm^3$ vs. $17.5{\pm}5.8%$, $1,377{\pm}436/mm^3$, each p=0.0001 and 0.002). The proportion of immature lymphocyte in the BM showed a decreasing trend in the FAPTB group($9{\pm}4%$ vs. $12{\pm}3%$, p=0.138). The IL-2($26.0{\pm}29.1$ vs. $112.2{\pm}42.4pg/mL$, p=0.001) and IL-10($3.4{\pm}4.7$ vs. $12.0{\pm}8.0pg/mL$, p=0.031) levels in the BM were significantly lower in the FAPTB group than those in control. The levels of the other cytokines in FAPTB group and control were similar. Conclusion : These results suggest that the cause of lymphocytopenia in PB is associated with a abnormality IL-2 and IL-10 production in the BM. More study will be needed to define the mechanism of a decreased reservoir in BM.

Inhibitory Effect of Cell Differentiation against 3T3-L1 Pre-Adipocytes and Angiotensin Converting Enzyme (ACE) Activity of Ice Plant (Mesembryanthemum crystallinum) (아이스플랜트(Mesembryanthemum crystallinum)의 3T3-L1 전지방세포 분화 및 Angiotensin-Converting Enzyme(ACE) 활성 억제)

  • Kang, Seung Mi;Kim, Seon Jeong;Nam, Sanghae
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.8
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    • pp.1012-1017
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    • 2017
  • Mesembryanthemum crystallinum (Family: Aizoaceae) is an annual plant consisting of ice crystal-shaped bladder cells, which is responsible for its common name ice plant. This study investigated biological activities according to general components and extraction solvent in order to examine the functionality of ice plant. The total content of free amino acids was 32.57 mg/g, including 4.64 mg/g of L-alanine as the most abundant and 2.60 mg/g of ${\gamma}$-aminobutyric acid. Regarding angiotensin converting enzyme inhibitory activities of solvent fractions of ice plant, ethyl acetate fraction and chloroform fraction showed activities of $33.17{\pm}3.20{\sim}88.19{\pm}3.20%$ and $23.72{\pm}2.89{\sim}86.78{\pm}2.24%$, respectively, similar to $Captopril^{(R)}$ ($19.51{\pm}3.44{\sim}84.72{\pm}1.06%$) and $Enalapril^{(R)}$ ($24.93{\pm}1.12{\sim}91.32{\pm}3.62%$) as positive control groups. Regarding inhibition of lipid droplet production in 3T3-L1 preadipocytes by ice plant, anti-adipogenic activities were $53.00{\pm}0.45{\sim}65.75{\pm}0.31%$ and $44.16{\pm}0.29{\sim}63.32{\pm}0.36%$ in the ethyl acetate fraction and butanol fraction, respectively, showing the lowest lipid droplet production. The chloroform fraction and hexane fraction showed activities of $38.33{\pm}0.09{\sim}56.55{\pm}0.50%$ and $31.17{\pm}0.50{\sim}55.10{\pm}1.93%$, respectively, whereas the water fraction showed activity of $26.32{\pm}2.27{\sim}49.48{\pm}0.05%$. Therefore, all solvent fractions inhibited fat accumulation of 3T3-L1 preadipocytes according to treatment concentration. According to the results above, it would be possible to utilize ice plant as a new health functional material.

Selective Expansion of TCR $V{\beta}3$+CD4+T Cells in Collagen-induced Arthritis in DBA/1 Mice (콜라겐 유도 관절염에서 콜라겐 항원 특이 $V{\beta}3$+CD4+T 세포의 선택적 증식)

  • Lee, Jae-Seon;Cho, Mi-La;Lee, Jung-Eun;Min, So-Youn;Yoon, Chong-Hyeon;Kim, Wan-Uk;Min, Jun-Ki;Park, Sung-Hwan;Kim, Ho-Youn
    • IMMUNE NETWORK
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    • v.5 no.2
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    • pp.78-88
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    • 2005
  • Background: Collagen-induced arthritis (CIA) in mice is animal model of autoimmune disease known as rheumatic arthritis in human. We investigated CII-specific CD4+ T cell receptor usage in CIA mice. Methods: In CIA model, draining lymph node (dLN) CD4+ T cells and splenocytes at $3^{rd},\;5^{th},\;8^{th}$ week, we investigated CII-specific T cell proliferation, production of IL-17, IFN-${\gamma}$, TNF-${\alpha}$, IL-4 and IL-10. And we also performed anti-CII IgG Ab measurements in serum level, TCRV ${\beta}$ usage and T cell clonality with RT-PCR-SSCP analysis. Also, we performed proliferative response against CII when CII-specific T cell subset is deleted. Results: CIA mice showed more increase in the serum level of anti-CII IgG than normal mice after induction of arthritis. And the level of anti-CII IgG2a in CIA mice was increased after $3^{rd}$ week after primary immunization, while anti-CII IgG1 was decreased. Draining LN CD4+ T cells have proliferated against CII stimulation at $3^{rd}$ week after $1^{st}$immunization. CD4+T cells derived from dLN of CIA mice produced proinflammatory cytokine IFN-${\gamma}$, IL-17 etc. Draining LN CD4 T cells of CIA presented higher proportion of CD4+V ${\beta}3$+subset compared to those of normal mice at $3^{rd}$ week after $1^{st}$ immunization, and they were increased in proportion by CII stimulation. Draining LN CD4+ T cells without TCRV ${\beta}3+/V{\beta}8.1/8.2+/V{\beta}$10b+cells were not responsive against CII stimulation. But, CII-reactive response of TCRV ${\beta}3-/V{\beta}8.1/8.2-/V{\beta}$10b- T cells was recovered when $V{\beta}3+$ T cells were added in culture. Conclusion: Our results indicate that CD4+$V{\beta}3+$ T cells are selectively expanded in dLN of CIA mice, and their recovery upon CII re-stimulation in vitro, as well as the production Th1-type cytokines, may play pivotal role in CIA pathogenesis.

Antioxidant and Anti-Adipogenic Effects of Ethanolic Extracts from Tartary and Common Buckwheats (쓴메밀 및 단메밀 에탄올 추출물의 항산화 및 지방세포 분화억제 효과)

  • Yoon, Bo-Ra;Cho, Bong-Jae;Lee, Hyo-Ku;Kim, Dae-Jung;Rhee, Seong-Kap;Hong, Hee-Do;Kim, Kyung-Tack;Cho, Chang-Won;Choi, Hyeon-Son;Lee, Boo-Yong;Lee, Ok-Hwan
    • Food Science and Preservation
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    • v.19 no.1
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    • pp.123-130
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    • 2012
  • In this study, 80% ethanolic extracts of tartary and common buckwheats were assessed for their total phenol content, total flavonoids content, antioxidant activity (DPPH, ABTS radical scavenging activity and reducing power), and anti-adipogenic effects. Our results show that total phenol contents of 80% ethanolic extract from tartary and common buckwheats were $17.35{\pm}0.41$ and $8.20{\pm}0.28\;{\mu}g$ GAE/g, respectively. Antioxidant activities of 80% ethanolic extract from tartary buckwheat were significantly higher than that of common buckwheat extract (p<0.05). During adipocyte differentiation, 80% ethanolic extracts of tartary and common buckwheat significantly inhibited lipid accumulation compared to control cells. We further evaluated the effect of buckwheat extracts on the changes of key gene expression associated with 3T3-L1 adipogenesis and ROS production. Tartary buckwheat extract was more suppressed the mRNA expressions ($PPAR{\gamma}$ and aP2) than that of common buckwheat extract. Moreover, tartary buckwheat inhibited the mRNA expression of both NOX4 (NADPH oxidase 4) and G6PDH (glucose-6-phosphate dehydrogenase). These results indicate that anti-adipogenesis effect of tartary buckwheat can be attributed to phenolic compound that may potentially inhibit ROS production.

Experimental Studies on the inflammation-related diseases pharmacological effect of water and 70% ethanol extracts from Socheongnyong-tang (소청룡탕(小靑龍湯) 물 및 에탄올 추출물의 염증 관련 질환 약리 효능에 관한 실험적 연구)

  • Jeon, Woo-Young;Lee, Mee-Young;Lim, Hye-Sun;Shin, In-Sik;Kim, Yeji;Jin, Seong Eun;Yoo, Sae-Rom;Seo, Chang-Seob;Kim, Jung-Hoon;Ha, Hyekyung;Jeong, Soo-Jin;Kim, Ohn Soon;Shin, NaRa;Kim, Seong-Sil;Shin, Hyeun-Kyoo
    • Herbal Formula Science
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    • v.20 no.2
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    • pp.13-28
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    • 2012
  • Objectives : The present study aimed to investigate the pharmacological activity of water and ethanol (EtOH) extracts from Socheongnyong-tang (SCNT) on inflammation and its related disease. Methods : The cells were treated with nontoxic concentrations of water and EtOH extract from SCNT in BEAS-2B, HaCaT, RAW 264.7 and 3T3-L1 cells. These cells were stimulated by tumor necrosis facter (TNF)-${\alpha}$, TNF-${\alpha}$/interferon (IFN)-${\gamma}$, and lipopolysaccharide (LPS), respectively. 3T3-L1 cells were differentiated by insulin. After incubation, supernatant were collected and biological indicator measured by enzyme-linked immunosorbent assay. Results : Our results indicate that the water and EtOH extract of SCNT significantly inhibited the production of regulated on activation normal T-cell expression and secreted (RANTES) by treatment of TNF-${\alpha}$ in BEAS-2B cell, and significantly reduced the production of RANTES and macrophage-derived chemokine increased by treatment of TNF-${\alpha}$/IFN-${\gamma}$ in HaCaT cell. Moreover, those extracts significantly decreased the activity of nitric oxide and prostaglandin $E_2$ in LPS-induced RAW 264.7, and significantly inhibited the increased activity of glycerol-3-phosphate dehydrogenase and expression of leptin induced by differentiation in 3T3-L1 cell. Conclusions : These results indicate that both water and EtOH extract of SCNT has powerful effects on inflammation and its related disease. Therefore, SCNT can be developed as a potential pharmacological agent related various diseases. Although the significant effects were observed in both SCNT water and EtOH extract, the EtOH extract was more effective on most experiments than its water extract. Taken together, these findings indicate that the SCNT EtOH extract may have more potential pharmacological agent.

Functional analysis of seaR protein identified from Saccharopolyspora erythraea (희소방선균의 seaR 단백질 발현을 통한 기능 분석)

  • Ryu, Jae Ki;Kwon, Pil-Seung;Lee, Hyeong Seon
    • Korean Journal of Microbiology
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    • v.51 no.1
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    • pp.39-47
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    • 2015
  • Secondary metabolism in actinomycetes has been known to be controlled by a small molecule, ${\gamma}$-butyrolactone autoregulator, the binding of which to each corresponding receptor leads to the regulation of the transcriptional expression of the secondary metabolites. We expected that expression of an autoregulator receptor or a pleiotropic regulator in a non-host was to be gained insight of effective production of new metabolic materials. In order to study the function of the receptor protein (seaR), which is isolated from Saccharopolyspora erythraea, we introduced the seaR gene to Streptomyces coelicolor A3(2) as host strains. An effective transformation procedure for S. coelicolor A3(2) was established based on transconjugation by Escherichia coli ET12567/pUZ8002 with a ${\varphi}C31$-derived integration vector, pSET152, which contained int, oriT, attP and $ermEp^*$ (erythromycin promotor). Therefore, the pEV615 was introduced into S. coelicolor A3(2) by conjugation and integrated at the attB locus in the chromosome of the recipients by the ${\varphi}C31$ integrase (int) function. Exconjugant of S. coelicolor A3(2) containing the seaR gene was confirmed by PCR and transcriptional expression of the seaR gene in the transformant was analyzed by RT-PCR. In case of S. coelicolor A3(2), a phenotype microarray was used to analyze the phenotype of transformant compared with wild type by seaR expression. After that, in order to confirm the accuracy of the results obtained from the phenotype microarray, an antimicrobial susceptibility test was carried out. This test indicated that sensitivity of the transformant was higher than wild type in tetracycline case. These results indicated that some biosynthesis genes or resistance genes for tetracycline biosynthesis in transformant might be repressed by seaR expression. Therefore, subsequent experiments, analysis of transcriptional pattern of genes for tetracycline production or resistance, are needed to confirm whether biosynthesis genes or resistance genes for tetracycline are repressed or not.

Rice Yield Estimation Using Sentinel-2 Satellite Imagery, Rainfall and Soil Data (Sentinel-2 위성영상과 강우 및 토양자료를 활용한 벼 수량 추정)

  • KIM, Kyoung-Seop;CHOUNG, Yun-Jae;JUN, Byong-Woon
    • Journal of the Korean Association of Geographic Information Studies
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    • v.25 no.1
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    • pp.133-149
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    • 2022
  • Existing domestic studies on estimating rice yield were mainly implemented at the level of cities and counties in the entire nation using MODIS satellite images with low spatial resolution. Unlike previous studies, this study tried to estimate rice yield at the level of eup-myon-dong in Gimje-si, Jeollabuk-do using Sentinel-2 satellite images with medium spatial resolution, rainfall and soil data, and then to evaluate its accuracy. Five vegetation indices such as NDVI, LAI, EVI2, MCARI1 and MCARI2 derived from Sentinel-2 images of August 1, 2018 for Gimje-si, Jeollabuk-do, rainfall and paddy soil-type data were aggregated by the level of eup-myon-dong and then rice yield was estimated with gamma generalized linear model, an expanded variant of multi-variate regression analysis to solve the non-normality problem of dependent variable. In the rice yield model finally developed, EVI2, rainfall days in September, and saline soils ratio were used as significant independent variables. The coefficient of determination representing the model fit was 0.68 and the RMSE for showing the model accuracy was 62.29kg/10a. This model estimated the total rice production in Gimje-si in 2018 to be 96,914.6M/T, which was very close to 94,470.3M/T the actual amount specified in the Statistical Yearbook with an error of 0.46%. Also, the rice production per unit area of Gimje-si was amounted to 552kg/10a, which was almost consistent with 550kg/10a of the statistical data. This result is similar to that of the previous studies and it demonstrated that the rice yield can be estimated using Sentinel-2 satellite images at the level of cities and counties or smaller districts in Korea.

Immunostimulatory and Anti-Obesity Activity of Lonicera insularis Nakai Extracts in Mouse Macrophages RAW264.7 Cells and Mouse Adipocytes 3T3-L1 Cells (섬괴불나무(Lonicera insularis Nakai) 추출물의 면역자극 및 항비만 활성)

  • Yu, Ju Hyeong;Yeo, Joo Ho;Choi, Min Yeong;Lee, Jae Won;Geum, Na Gyeong;An, Mi-Yun;Jeong, Jin Boo
    • Korean Journal of Plant Resources
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    • v.35 no.4
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    • pp.417-427
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    • 2022
  • In this study, we investigated in vitro immuno-stimulatory and anti-obesity activity of fruit (LIF), leaves (LIL) and stems (LIS) from Lonicera insularis Nakai in mouse macrophages RAW264.7 cells and mouse pre-adipocytes 3T3-L1 cells. LIF, LIL and LIS increased the production of immunostimulatory factors such as nitric oxide (NO), inducible nitric oxide synthase (iNOS), interleukin-1β (IL-1β), cyclooxygenase-2 (COX-2), tumor necrosis factor-α (TNF-α) and activated phagocytosis in RAW264.7 cells. Inhibition of toll-like receptor 2/4 (TLR2/4) partly blocked LIF, LIL and LIS mediated production of immunostimulatory factors. In addition, inhibition of mitogen-activated protein kinases (MAPK) signaling attenuated the production of immunostimulatory factors induced by LIF, LIL and LIS. Based on these results of this study, LIF, LIL and LIS is thought to activate macrophages the production of immunostimulatory factors and phagocytosis through toll-like receptor 2/4 (TLR2/4) and MAPKs signaling pathway. In anti-obesity study, LIF reduced the lipid accumulation in 3T3-L1 cells. LIF increased the protein phosphorylation expressions such as AMP-activated protein kinase (AMPK), hormone sensitive lipase (HSL), adipose triglyceride lipase (ATGL) related to the lipolysis of the adipocytes. In addition, LIF increased the expression of proteins involved in energy metabolism and brown adipose tissues differentiation such as peroxisome proliferator-activated receptor gamma coativator 1α (PGC-1α) and PR domain-containing16 (PRDM16). These results suggest that LIF is involved in lipid accumulation inhibition through expressing the proteins such as lipolysis and differentiation of white adipocytes to brown adipocytes.