• 제목/요약/키워드: C57BL%2F6

검색결과 70건 처리시간 0.025초

Jl 배아주세포를 이용한 효율적인 생식선 이행 카이미라의 생산 (Factors Affecting the Productivity of Germ-line Chimeras from Jl Embryonic Stem Cells)

  • 김선욱;구본실;정상균;이태훈;유성란;남윤이;김정림;현병화;신희섭;이경광;상병찬;유대열
    • 한국가축번식학회지
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    • 제25권1호
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    • pp.71-77
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    • 2001
  • 본 실험은 phospholipase C (PLC)-$\beta$-3 및 peroxiredoxin (Prx) II 유전자가 적중 ($\Delta$)된 J1 마우스 배아주 (embryonic stem) 세포로부터 생식선이행 카이미라 마우스생산을 위한 제반조건 및 배아주세포의 배양조건을 확립하기 위하여 수행되었다. 80% 이상의 정상핵형을 보이는 유전자 적중된 4개의 클론 ($\Delta$Pu II C3, $\Delta$Pu II C3, C10 및 15)으로부터 카이미라를 생산하였을 때 형태적으로 분화정도가 높은 클론 ($\Delta$PLC$\beta$-3 C3)의 이용은 카이미라의 생산율 (21.1%)과는 무관하게 카이미리즘 (< 20%)이 낮았고, 수컷 카이미라의 생산 빈도 (7/15, 46%)도 낮아지는 것으로 나타났다. 그러나 형태적으로 안정된 3개의 클론 ($\Delta$Prx II C3, C10 및 15)은 80% 이상의 높은 카이미리즘을 지닌 마우스 (9/13, 69.2%)를 생산하였고, 수컷 카이미라의 생산율 (l1/13, 84.6%)도 증대된 것으로 나타났다. 따라서 80% 이상의 정상핵형을 지닌 배아주 세포를 형태적으로 안정하게 유지하는 것이 카이미리즘이 높은 마우스를 생산하는데 결정적 요인으로 작용할 수 있는 것으로 확인되었다. 미세주입용 배반포를 효율적으로 생산하기 위해 5~10주령 사이의 C57BL/6J 암컷마우스를 교배 한 결과, 10주령 마우스가 미세주입가능한 3.5일령 배반포를 가장 많이 생산 (2.94개/마리)하였다. 또한 미세주입된 배반포를 이식하기 위해 ICR 및 ICR$\times$C57BL/6J F1 (IBF1)위임신 대리모를 사용하였을 때, IBF1이 복당 산자수 (2.8 vs. 5.6)가 많았고 카이미라 생산율 (0 vs. 35.3%)도 매우 높았다. 따라서 공여마우스의 주령 및 대리모의 선택이 카이미라 생산효율 향상에 중요한 요인으로 부각되었다. 결과적으로 핵형이 안정된 ES 세포를 동정하는 것은 물론 클로닝 과정중에 형태적으로 분화가 없도록 ES세포를 배양하는 것이 카이미리즘이 높은 마우스를 생산하고 아울러 생식선 이행 빈도를 증가시키는데 결정적인 역할을 하는 것으로 확인되었다.

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마우스 동종 줄기세포 유래 수지상 세포를 이용한 백신의 흑색종 폐암 전이 모델에서의 항암 효과 및 기전 연구 (Anti-cancer Effect of Hematopoietic Stem Cell-derived Allogeneic-DC Vaccine in Melanoma Metastasis Model)

  • 김명주;손혜진;백소영;이강은;이영준;이현아
    • IMMUNE NETWORK
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    • 제6권3호
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    • pp.154-162
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    • 2006
  • Background: Dendritic cell (DC)-based cancer immunotherapy is studied for several years. However, it is mainly derived from autologous PBMC or leukapheresis from patient, which has limitations about yield and ability of DC production according to individual status. In order to solve these problems, inquiries about allogeneic DCs are performed but there are no preclinical trial answers for effect or toxicity of allogeneic DC to use for clinical trial. In this study, we compared the anti-tumor effect of allogeneic and autologous DCs from mouse bone marrow stem cells in mouse metastatic melanoma model. Methods: B16F10 melanoma cells ($5{\times}10^4$/mouse) were injected intravenously into the C57BL/6 mouse. Therapeutic DCs were differentiated from autologous (C57BL/6: CDC) or allogeneic (B6C3F1: BDC) bone marrow stem cells with GM-CSF, SCF and IL-4 for 13days and pulsed with B16F10 tumor cell lysate (Blys) for 18hrs. DC intra-peritoneal injections began on the 8th day after the tumor cell injection by twice with one week interval. Results: Anti-tumor response was observed by DC treatment without any toxicity especially in allogeneic DC treated mice (tumor burden score: $2.667{\pm}0.184,\;2.500{\pm}0.463,\;2.000{\pm}0.286,\;1.500{\pm}0.286,\;1.667 {\pm}0.297$ for saline, CDC/unpulsed-DC: U-DC, CDC/Blys-DC, BDC/U-DC and BDC/Blys-DC, respectively). IFN-${\gamma}$ secretion was significantly increased in allogeneic DC group stimulated with B16F10 cell lysate ($2,643.3{\pm}5,89.7,\;8,561.5{\pm}2,204.9.\;6,901.2{\pm}141.1pg/1{\times}10^6$ cells for saline, BDC/U-DC and BDC/Blys-DC, respectively) with increased NK cell activity. Conclusion: Conclusively, promising data was obtained that allogeneic DC can be used for DC-based cancer immunotherapy.

항암(抗癌) 및 면역효과(免疫效果)에 복분자약침(覆盆子藥鍼)이 미치는 영향(影響) (Study on the Anti-cancer, Anti-metastasis and Immune response improvement Effects of Aqua-acupuncture with Rubi Fructus infusion solution)

  • 이선희;이병렬
    • Journal of Acupuncture Research
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    • 제17권1호
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    • pp.189-219
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    • 2000
  • To study the effects of anti-cancer, anti-metastasis and immune response improvement effects of aqua-acupuncture with Rubi Fructus infusion solution, we used Rubi Fructus infusion solution(taken by water-alcohol method) put into Chung-wan (CV12) and Chok-Samni(ST36) of BALB/c or C57BL/6 which are corresponding to humanbody. We observed the cytotoxicity, the effect on the expression of MMP-9 gene, the ability to control cancer cell proliferation, change of body weight, surviving number, median surviving time, increase of life span, changes in amount of leukocyte, erythrocyte, platelet, total protein, creatinine, glucose and LDH, weight of spleen, number of pulmonary colony, histological analysis on tissue metastasis of lung and liver, splenic cell proliferation, the expression of cytokine gene, the number of $CD4^+$, $CD8^+$, $CD19^+$ and NK cell, and concluded like this. The results were obtained as follows : 1. Effects of Anti-cancer 1) The cytotoxicity about B16-F10 cell line of $2^0$, $2^{-1}$, $2^{-2}$, $2^{-3}$, $2^{-5}$, $2^{-6}$, $2^{-7}$, $2^{-8}$ diluent groups in Rubi Fructus infusion solution treatment was inhibited significantly, compared with control group. 2) The cytotoxicity about HT1080 cell line of $2^0{\sim}2^{-8}$ diluent groups in Rubi Fructus infusion solution treatment was inhibited significantly, compared with control group. 3) The effect on expression of MMP-9 gene was inhibited significantly in all the sample groups, compared with control group. 4) The effect on the control-ability on the cancer cell proliferation showed cytotoxicity significantly in $2^0$, $2^{-1}$, $2^{-2}$, $2^{-3}$, $2^{-4}$, $2^{-5}$, $2^{-6}$, $2^{-7}$, diluent groups. 2. Effects of Anti-metastasis 1) S-180 cancer cell line transplants in BALB/c mice were inhibited significantly in weight increase in all the sample groups, compared with control group. The surviving number increased in almost sample groups, except one group put into Chok-Samni(ST36) with 20% Rubi Fructus infusion solution treatment group that showed same number of the control group. 2) S-180 cancer cell line transplants in BALB/c mice showed high MST significantly in almost sample groups, compared with control group. But one group put into Chok-Samni(ST36) with 20% Rubi Fructus infusion solution showed low MST than control group. 3) The group injected in vein with B16-F10 cancer cell line in C57BL/6 mice showed increased ILS than control group significantly in anti-metastasis test. 3. Effects of Immune response improvement 1) The group injected in vein with B16-F10 cancer cell line in C57BL/6 mice were increased significantly in the number of leukocyte and glucose, and decreased significantly in the amount of platelet and LDH, compared with control group. However, there's no significant increase or decrease in number of erythrocyte, total protein and creatinine. 2) We couldn't find any significant relation in spleen weight of the sample group. 3) In pulmonary colony, sample group was decreased significantly, compared with control group. 4) Histological analysis of sample group inhivited compared with that of control group in both of lung and liver. 5) In immune system, all the sample groups showed having more relevancy to the effect on splenic cell proliferation than normal group. 6) Cytokine gene increased in almost sample groups, except one group treated with $50{\mu}g/m{\ell}$ Rubi Fructus infusion solution on IL-12. 7) In flow cytometry there's no significant relation in number of $CD8^+$ cell, however, the number of $CD4^+$, $CD19^+$ cell and NK cell in sample group had more relation than in control group. Above the results showed that aqua-acupuncture of Rubi Fructus solution has effects of anti-cancer, and-metastasis and immune response improvement.

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육종용약침의 항암작용(抗癌作用) 및 면역효과(免疫效果)에 대(對)한 실험적(實驗的) 연구(硏究) (Study on the Anti-cancer, Anti-metastasis and Immune response improvement of Aqua-acupuncture with Cistanches Herba infusion solution)

  • 이은용;이병렬
    • Journal of Acupuncture Research
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    • 제17권1호
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    • pp.251-286
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    • 2000
  • To study the effects of anti-cancer, anti-metastasis and immune response improvement of aqua-acupuncture with Cistanches Herba infusion solution, we used Cistanches Herba infusion solution(taken by water-alcohol method) put into Chung-wan(CV12) and Chok-Samni(S36) of BALB/c or C57BL/6 which are corresponding to human body. We observed the cytotoxicity, the effect on the expression of MMP-9 gene, the ability to control cancer cell proliferation, change of body weight, surviving number, MST, ILS, changes in amount of WBC, RBC, PLT, total protein, creatinine, glucose and LDH, weight of spleen, number of pulmonary colony, histological analysis on tissue metastasis of lung and liver, splenic cell proliferation, the expression of cytokine gene, the number of $CD4^+$, $CD8^+$, $CD19^+$ and NK cell, and concluded like this. The results were obtained as follows : 1. The cytotoxicity about B16-F10 cell line of $2^0$, $2^{-1}$, $2^{-2}$, $2^{-5}$ diluent groups in Cistanches Herba infusion solution treatment was inhibited significantly, compared with control group. 2. The cytotoxicity about HT1080 cell line of $2^0$, $2^{-1}$, $2^{-2}$, $2^{-3}$, $2^{-5}$, $2^{-8}$ diluent groups in Cistanches Herba infusion solution treatment was inhibited significantly, compared with control group. 3. The effect on expression of MMP-9 gene was decreased in all the sample groups, compared with control group. 4. The effect on the control-ability on the cancer cell proliferation showed cytotooicity significantly in $2^0$, $2^{-1}$, $2^{-2}$, $2^{-3}$, $2^{-4}$, $2^{-5}$ diluent groups. 5. S-180 cancer cell line transplants in BALB/c mice were inhibited significantly in weight increase in all the sample groups, compared with control group. The surviving number increased in almost sample groups, except one group put into Chok-Samni(S36) with 20% Cistanches Herba infusion solution treatment group that showed same number of the control group. 6. S-180 cancer cell line transplants in BALB/c mice showed high MST and ILS significantly in almost sample groups, compared with control group. But one group put into Chok-Samni(S36) with 20% Cistanches Herba infusion solution treatment group showed low MST and ILS than control group. 7. The sample group injected in vein with B16-F10 cancer cell line in C57BL/6 mice showed increased ILS compared with control group significantly in anti-metastasis test. 8. The sample group injected in vein with B16-F10 cancer cell line in C57BL/6 mice were increased significantly in the number of WBC and glucose, and decreased significantly in the amount of LDH, compared with control group. However, there's no significant increase or decrease in number of RBC, PLT, total protein and creatinine. 9. We couldn't find any significant relation in spleen weight of the sample group. 10. In pulmonary colony, sample group was decreased significantly, compared with control group. 11. Histological analysis of sample group inhivited compared with that of control group in both of lung and liver. 12. In immune system, all the sample groups showed having more relevancy to the effect on splenic cell proliferation than normal group. 13. The effect on cytokine gene expression of all the sample groups were increased than control group. 14. In flow cytometry there's no significant relation in number of $CD8^+$, $CD19^+$ cell, however, the number of $CD4^+$ cell and NK cell in sample groups were increased than in control group. Above the results showed that aqua-acupuncture of Cistanches Herba infusion solution has effects of anti-cancer, anti-metastasis and immune response improvement.

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Anti-tumor Activity of the Fruitbody Extract of Basidiomycete, Phellinus linteus

  • Lim, Jong-Soon;Kim, Seung-Hyung;Park, Jin-Seo;Park, Jeong-Youl;Park, Seong-Joo;Shin, Kwang-Soo
    • Journal of Microbiology
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    • 제39권2호
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    • pp.121-125
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    • 2001
  • Methanol extract prepared from the fruitbody of Phellinus linteus (EPL) showed anti-tumor and immune-stimulating activities. The invasion activity of Bl6-F10 melanoma cells through a reconstituted basement membrane to the collagen-coated lower surface of the filters was inhibited about 67% by EPL (100 $\mu\textrm{g}$/ml). Also, EPL inhibited the expression of the mRNA for MMP-2 and MMP-9. In vivo treatment of C57BL/6 mice (150 mg/kg) with EPL for 14 days, the pulmonary colonization was found to be inhibited about 75%. Using reverse transcriptionpolymerase chain reaction (RT-PCR) analysis, we found that cytokine IL-12 and INF-${\gamma}$ genes were induced by EPL. Furthermore, EPL stimulated the proliferation of CD4$\^$+/(33.5%) and CD8$\^$+/(17.7%) in mouse splenocytes.

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Infection Characteristics of Korean Trichinella Isolate to Some Kind of Experimental Animals

  • 손운목
    • 대한의생명과학회지
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    • 제12권2호
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    • pp.99-104
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    • 2006
  • Present study was performed to investigate the host-parasite relationship of the Korean Trichinella isolate (KTI). In the experiment to observe the infectivity of KTI to several kinds of animals, the reproductive capacity index (RCT) was highest in cats, and that in mice, hamsters and rats was followe4 in descending order. However, birds, i.e. wild goose and chicken, did not infect with KTI. The number of larvae per a gram of muscle (LPG: 377) was highest in the tongue of cats experimentally infected with KTI larvae. LPG in the diaphragm, anterior leg, back, posterior leg and abdominal muscles were 313, 246, 234, 225 and 170 respectively. Muscle larvae recovered at 55 days after infection were revealed the highest infectivity (RCI: 137.2) in mice. RCI was comparatively low in the mice infected with less than 25 day-old and more than 300 day-old larvae. In the experiment to observe the susceptibility of KTI by the mouse strain, ICR (RCI: 137.2), C57BL/6 (RCI: 108.8), DBA/2 (RCI: 107.1), C3H (RCI: 98.7), BALB/c (RCI: 96.9), FVB (RCI: 96.1) and B6C3F1 (RCI: 85.3) were very susceptible. However, BDF1 (RCI: 57.7) and CBA (RCI. 57.1) were revealed the moderate susceptibility, and B6CBAF1 (RCI: 23.1) was shown the lowest. The infection sites of adults were posteriorly transferred in the small intestine of experimental mice according to the infection periods of muscle larvae. The infection characteristics of KTI observed in this study may be useful as the basic data in the advanced studies, furthermore in the study of other Trichinella isolates.

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순기제습방(順氣除濕方)이 고지방식이(高脂肪食餌)로 유발된 비만(肥滿) 백서(白鼠)에 미치는 영향 (The inhibitory effects of Soongijeseub-bang(順氣除濕方) on the obese-mice caused by high-fat diet)

  • 이우열;신현규
    • 한국한의학연구원논문집
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    • 제14권1호
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    • pp.161-171
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    • 2008
  • Objective : In order to investigate the effects of Soongijeseub-bang(here in after referred to SJB) on the obese gene and obese inhibitory, C57BL/6 mice were induced by high fat diet Methods : C57BL/6 mice were divided into three groups(normal, high fat diet with control, high fat diet with SJB extract) and fed for 15 weeks. And measured body weight change, the weight change of the adipocytes in body, the levels of ALT, AST, creatinine, total cholesterol, LDL-cholesterol, HDL-cholesterol, triglyceride, NEFA(non-esterified fat acid), glucose in serum and the expression of ${\beta}3AR$, leptin, and TNF- ${\alpha}$ gene in primary adipocytes and adipocytes tissue. Results : The following results were obtained in this study. 1. SJB 500 mg/kg extract group showed significant decrease in final weight. 2. SJB 500 mg/kg extract group showed significant decrease in the amount of adipocyte in weight. 3. All experimental group showed significant decrease in ALT and AST levels. 4. All experimetal group did not show significant decrease in glucose and creatinine levels. 5. All experimental group showed significant decrease in total cholesterol, LDL-cholesterol, triglyceride and NEFA levels. HDL-cholesterol were increase significantly in SJB 500 mg/kg extract group. 6. SJB 500 mg/kg extract group showed significant decrease in leptin levels. 7. All experimental group showed significantly increased expressions of fJ 3AR in primary adipose cell and 3T3-Ll cell, and those of leptin in primary adipose cell and 3T3-Ll cell were decreased significantly. SJB $100{\mu}l/ml$ extract group showed significant decrease in the expression of TNF-${\alpha}$. 8. SJB 500 mg/kg extract group showed decrease in the size of adipocyte in adipocytes tissue. 9. All experimental group showed significant decrease in adipose vacuoles in liver tissue.

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발아 서목태 가수분해물의 근위축 억제 효과 (Germinated Rhynchosia nulubilis Hydrolysate Ameliorates Dexamethasone-induced Muscle Atrophy by Downregulating MAFbx Expression in C2C12 Cells and C57BL/6 Mice)

  • 이원경;김은지;김상곤;구영민;길영숙;신승미;안민주;강민철;하영술
    • 생명과학회지
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    • 제33권3호
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    • pp.277-286
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    • 2023
  • 스테로이드 유발 근위축은 장기간 코르티코스테로이드 요법의 일반적인 부작용이며 근육량과 기능의 현저한 감소로 이어질 수 있다. 발아된 Rhynchosia Nulubilis (GRH)는 항염증 및 항산화 특성이 있는 것으로 밝혀진 전통적인 약용 식물이다. 그러나 근 감소 억제 효능에 대하여는 보고된 바가 없다. 본 연구에서는 덱사메타손에 의해 유도된 근위축 C2C12 근관세포 모델과 마우스 모델에서 근위축 억제 활성에 대한 GRH의 영향을 평가하였다. 또한 골격근에 대한 GRH 효과의 기본이 되는 분자 경로를 확인하였다. May Grunwald-Giemsa 염색을 통한 근관 세포 염색 결과는 GRH 처리군에서 근관의 길이와 면적이 증가함을 보여주었다. 또한, muscle ring-finger protein-1 (MuRF-1)와 muscle atrophy F-box (MAFbx) 발현이 GRH 처리군에서 유의하게 감소하였다. 더욱이 덱사메타손에 의해 유도된 근위축 C57BL/6 마우스에서 트레드밀을 사용한 지구력 측정과 악력측정기를 이용한 악력 측정을 통한 근육 기능을 평가한 결과 GRH를 경구 투여한 마우스의 근육 기능이 향상되었다. 또한 이러한 결과는 근육 조직에서 GRH에 의한 MAFbx의 발현 억제를 통해 일어남을 확인하였다. 결론적으로 GRH는 MAFbx 발현을 억제하여 유비퀴틴-프로테아좀 경로를 차단함으로써 덱사메타손에 의해 유발된 근위축을 개선할 수 있을 것이다.

Defining B Cell Epitopes of Ovalbumin for the C57BL/6 Mice Immunized with Recombinant Mycobacterium smegmatis

  • Kim, Hyo-Joon;Lee, Yang-Min;Hwang, Joon-Sung;Won, Ho-Shik;Kim, Bok-Hwan
    • BMB Reports
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    • 제32권5호
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    • pp.461-467
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    • 1999
  • Recombinant Mycobacterium smegmatis expressing ovalbumin was used to immunize C57BL/6(H-$2^b$) mice, and the humoral immunity against recombinant ovalbumin was analyzed. Antibodies were purified by denatured ovalbumin-conjugated affinity chromatography. The epitopes of the antibodies were screened with a random peptide library displayed on the tip of fUSE5 filamentous phage pIII minor coat proteins. Two peptides, IRLADR and SPGAEV, were selected predominantly by the recognition of purified antibodies using biopanning methods. The composition of the peptide sequence with the primary structure of OVA revealed that the peptide sequence analogizes to INEAGR, part of the $^{323}ISQAVHAAHAEINEAGR^{339}$ sequence previously reported as the antigenic determinant for murine Band also Th cell epitopes (I-$A^d$ binding). Also, the structures of these mimotopes obtained from restrained molecular dynamic computations resulted in the formation of a $\beta$-turn proven to be a secondary structure of the parent peptide within the ovalbumin molecule, enabling us to confirm the structural similarity. This study demonstrates that immunization with recombinant M. smegmatis can generate neutralizing antibodies identical with those induced by the administration of natural antigenic proteins and supports the potential use of mycobacteria as vaccine delivery vehicles.

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TM-25659-Induced Activation of FGF21 Level Decreases Insulin Resistance and Inflammation in Skeletal Muscle via GCN2 Pathways

  • Jung, Jong Gab;Yi, Sang-A;Choi, Sung-E;Kang, Yup;Kim, Tae Ho;Jeon, Ja Young;Bae, Myung Ae;Ahn, Jin Hee;Jeong, Hana;Hwang, Eun Sook;Lee, Kwan-Woo
    • Molecules and Cells
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    • 제38권12호
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    • pp.1037-1043
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    • 2015
  • The TAZ activator 2-butyl-5-methyl-6-(pyridine-3-yl)-3-[2'-(1H-tetrazole-5-yl)-biphenyl-4-ylmethyl]-3H-imidazo[4,5-b]pyridine] (TM-25659) inhibits adipocyte differentiation by interacting with peroxisome proliferator-activated receptor gamma. 1 TM-25659 was previously shown to decrease weight gain in a high fat (HF) diet-induced obesity (DIO) mouse model. However, the fundamental mechanisms underlying the effects of TM-25659 remain unknown. Therefore, we investigated the effects of TM-25659 on skeletal muscle functions in C2 myotubes and C57BL/6J mice. We studied the molecular mechanisms underlying the contribution of TM-25659 to palmitate (PA)-induced insulin resistance in C2 myotubes. TM-25659 improved PA-induced insulin resistance and inflammation in C2 myotubes. In addition, TM-25659 increased FGF21 mRNA expression, protein levels, and FGF21 secretion in C2 myotubes via activation of GCN2 pathways (GCN2-$phosphoelF2{\alpha}$-ATF4 and FGF21). This beneficial effect of TM-25659 was diminished by FGF21 siRNA. C57BL/6J mice were fed a HF diet for 30 weeks. The HF-diet group was randomly divided into two groups for the next 14 days: the HF-diet and HF-diet + TM-25659 groups. The HF diet + TM-25659-treated mice showed improvements in their fasting blood glucose levels, insulin sensitivity, insulin-stimulated Akt phosphorylation, and inflammation, but neither body weight nor food intake was affected. The HF diet + TM-25659-treated mice also exhibited increased expression of both FGF21 mRNA and protein. These data indicate that TM-25659 may be beneficial for treating insulin resistance by inducing FGF21 in models of PA-induced insulin resistance and HF diet-induced insulin resistance.