博碩士論文 982204001 詳細資訊




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姓名 湯雅筑(Ya-Chu Tang)  查詢紙本館藏   畢業系所 生命科學系
論文名稱 第一型內皮素刺激3T3-L1脂肪細胞內抗胰島素激素基因的表現
(Endothelin-1 up-regulates resistin gene expression in 3T3-L1 adipocytes)
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摘要(中) 已有研究指出,抗胰島素激素 (resistin 和第一型內皮素(endothelin-1) 會抑制脂肪分化以及與脂肪細胞的胰島素抗性有關。 最近的研究也指出,抗胰島素激素會刺激第一型內皮素表現;第一型內皮素會刺激抗胰島素激素分泌,而此作用具有時間的效應。但第一型內皮素刺激抗胰島素基因表現的作用機制目前仍不清楚。本篇研究中,我們以3T3-L1脂肪細胞中首度討論第一型內皮素會刺激抗胰島素激素mRNA表現的訊息路徑。第一型內皮素所促進的抗胰島素mRNA表現,具有劑量與時間效應。100 nM 的第一型內皮素的作用下,0.25~12 小時所促進的抗胰島素mRNA表現約為100~250%。而前處理Actinomycin D則會抑制了第一型內皮素所增加的抗胰島素mRNA表現,此結果支持了第一型內皮素作用需要新mRNA的合成。由於,第一型內皮素並未改變在Actinomycin D作用之下的抗胰島素激素mRNA半衰期,所以推論第一型內皮素並不是透過改變mRNA穩定性而增加抗胰島素激素mRNA的表現量。前處理A型內皮素受器 ETAR 抑制劑,如BQ610,拮抗了第一型內皮素所增加的抗胰島素激素mRNA表現和下游訊息分子的磷酸化,例如ERK1卅2、JNKs、AKT和 STAT3等。但B型內皮素受器 (ETBR)抑制劑,如BQ788,並沒有這樣的效果。此外,分別前處理如ERK1卅2 (U0126 and PD98059), JNKs (SP600125), PI3K卅AKT (LY294002 and wortmannin), or JAK2卅STAT3 (AG490)的抑制劑,抑制了內皮素所促進的抗胰島素激素mRNA表現,並且分別降低第一型內皮素所刺激的訊息分子磷酸化,如ERK1卅2、JNKs、AKT 和STAT3。然而,p38激酶抑制劑並未抑制第一型內皮素的作用。上述結果顯示,第一型內皮素刺激抗胰島素激素基因是透過ETAR,以及ERK1卅2、JNKs、AKT和STAT3的活化,但ETBR和p38則不是必需的。
摘要(英) Resistin and endothelin (ET)-1 have been reported to inhibit adipogenesis and
regulate adipocyte insulin resistance, respectively. Recent studies have
demonstrated that resistin stimulated ET-1 expression and that ET-1
time-dependently stimulated resistin secretion, but the exact signaling pathway of
ET-1 to act on resistin gene expression is still unknown. In this study, we
investigated the signaling pathways involved in ET-1-stimulated resistin gene
expression in 3T3-L1 adipocytes. ET-1 up-regulated resistin mRNA expression in
dose- and time-dependent manners. The concentration of ET-1 that increased
resistin mRNA levels by 100-250% was approximately 100 nM after 0.25~12 h.
Treatment with actinomycin-D blocked ET-1-increased resistin mRNA levels,
suggesting that the effect of ET-1 requires new mRNA synthesis. Because ET-1 did
not alter the basal half-life of resistin mRNA induced by acti-D alone,
ET-1-stimulated resistin expression is unlikely due to through altered mRNA stability.
Treatment with an inhibitor of endothelin type A receptor (ETAR), such as BQ610,
but not with ETBR antagonist BQ788, blocked ET-1-increased levels of resistin
mRNA and phosphorylated levels of downstream signaling molecules, such as
ERK1/2, JNKs, AKT, and STAT3. Moreover, pre-treatment of specific inhibitors of
either ERK1/2 (U0126 and PD98059), JNKs (SP600125), PI3K/AKT (LY294002 and
wortmannin), or JAK2/STAT3 (AG490) prevented ET-1-increased levels of resistin
mRNA and respectively reduced ET-1-stimulated phosphorylation of ERK1/2, JNKs,
AKT and STAT3. However, p38 kinase antagonist SB203580 did not block the effect
of ET-1. These results suggest that ETAR, ERK1/2, JNKs, AKT, and STAT3, but not
ETBR or p38, are necessary for the ET-1 stimulation of resistin gene expression.
關鍵字(中) ★ 第一型內皮素
★ 3T3-L1脂肪細胞
★ 抗胰島素激素
關鍵字(英) ★ Endothelin-1
★ resistin
★ 3T3-L1 adipocyte
論文目次 中文摘要.........................i
Abstract....................ii
Declaration................iii
Acknowledgments.............iv
Contents.....................v
List of Tables.............vii
List of Figures...........viii
Abbreviations...............xi
Introduction..............1-13
Resistin.....................1
 1. The discovery............1
 2. Molecule structure.......2
 3. Distribution and physiological fucntions...............3
 4. Regulatoin...............7
Endothelin-1................10
 1. The discovery and isofors...........................10
 2. ET receptors............10
 3. Physiological functions.11
 4. Effects on adipokine expression and secretion.........13
Objectives..................14
Materials and Methods....15-22
 Materials..................15
 Cell culture...............16
 RNA extraction.............18
 Polymerase chain reaction (PCR)..........................18
 Western blot analysis......20
 Statistical analysis.......21
Results..................23-28
 Effects of ET-1 on resistin mRNA expression..............23
 ET-1 increases phosphorylation of ERK1卅2, JNKs, AKT and STAT3 proteins.................23
 ET-1 acts through the ETAR to increase resistin mRNA levels...........................24
 ERK and JNK MAPKs are involved in ET-1-stimulated expression of adipocyte resistin mRNA25
 ET-1-induced up-regulation of resistin mRNA expression depends on the PI3K卅AKT pathway.26
 The JAK2卅STAT3-regulated pathway mediates the ET-1 stmulation of resistin mRNA expression...........................27
Discussion..................29
Conclusions.................35
References..................36
Tables......................46
Figures.....................48
Appendix.................60-76
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指導教授 高永旭(Yung-Hsi Kao) 審核日期 2012-5-31
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