HORMONAL REGULATION OF HEPATIC AND PANCREATIC ISLET GLUCOKINASE mRNA LEVEL

肝脏和胰腺胰岛葡萄糖激酶 mRNA 水平的激素调节

• 批准号: 02671092
• 负责人: NISHI Shigeo
• 金额: $ 1.28万
• 依托单位: HAMAMATSU UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02671092/
• 关键词: Pancreatic islet glucokinase; Mechanism of gene regulation; competitive PCR; cAMP; cGMP; glibenclamide; 蛋白合成阻害; 膵ラ氏島グルコキナ-ゼ; ホルモンによる遺伝子発現調節; competitive PCR; cyclic AMP; グルコキナ-ゼ; 分子生物学

Measurement of islet glucokinase mRNA by competitive PCRSince it is too small amounts in total RNA extracted from isolated rat pancreatic islets to measure the abandance of glucokinase mRNA by Northern blot analysis, competitive PCR method was applied. This method enabled us to measure the level of glucokinase mRNA using as small as 0.2 ug of total RNA.Mechanism of regulation of islet glucokinase mRNAAfter 24 hs of preculture, isolated rat islats were cultured for additional 24 hs in RPMI medium (+10% FCS) with 2.2,5.5, or 16.7 mM glucose. In order to investigate the effects of verious hormones on the abandance of islet glucokinase mRNA, dexamethason (1uM), tri-iodothyronine (10uM), Glucagon (1uM), or insulin (1uM) was added in the culture medium. Islet glucokinase mRNA level was higher in the presence of 16.7 mM glucose than at 2.2 or 5.5 mM glucose. While administration of dexamethason, T3, and glucagon significantly inhibited the abandance of islet glucokinase mRNA, insulin treatment did not change glucokinase mRNA level. Cyclic-AMP analogue (dibutytyl cAMP) significantly reduced glucokinase mRNA level in 5.5 and 16.7 mM glucose although cyclic GMP analogue (8-bromo cGMP) produced no change in glucokinase mRNA. Furthermore, glucokinase mRNA increased at 6 hs after administration of oral hypoglycemic agent (glibenclamide), whereas glucokinase mRNA level decreased to basal values at 24 hs. These results suggest that various hormones and cyclic nucleotides regulate glucokinase mRNA level at pancreatic islets by the different mechanism from at hepatocytes.

竞争性PCR法检测胰岛葡萄糖激酶mRNA由于从大鼠胰岛中提取的总RNA量太少，不能用北方印迹法检测葡萄糖激酶mRNA的丰度，故采用竞争性PCR法。这种方法使我们能够使用小至0.2 μ g的总RNA来测量葡萄糖激酶mRNA的水平。胰岛葡萄糖激酶mRNA的调节机制预培养24小时后，将分离的大鼠胰岛在含有2.2、5.5或16.7mM葡萄糖的RPMI培养基（+10%FCS）中再培养24小时。为了研究各种激素对胰岛葡萄糖激酶mRNA表达的影响，在培养基中加入地塞米松（1 μ M）、三碘甲腺原氨酸（10 μ M）、胰高血糖素（1 μ M）或胰岛素（1 μ M）。胰岛葡萄糖激酶mRNA水平在16.7 mM葡萄糖的存在下高于2.2或5.5 mM葡萄糖。地塞米松、T3和胰高血糖素显著抑制了胰岛葡萄糖激酶mRNA的表达，而胰岛素治疗没有改变葡萄糖激酶mRNA的水平。环AMP类似物（二丁基cAMP）显着降低葡萄糖激酶mRNA水平在5.5和16.7 mM葡萄糖，虽然环GMP类似物（8-溴cGMP）产生的葡萄糖激酶mRNA没有变化。此外，口服降糖药（格列本脲）后6 h葡萄糖激酶mRNA水平升高，而24 h葡萄糖激酶mRNA水平降至基础值。这些结果表明，各种激素和环核苷酸调节葡萄糖激酶mRNA水平在胰岛通过不同的机制在肝细胞。

项目成果

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S.Nishi 等人：“人类胰腺 B 细胞 glacokinase：cDNA 序列和多态性基因对 chromorose7 bmcl p13 的定位。”

S.Hinata,S.Nishi,et al.: "Hormonal Regulation of Rat Pancreatic Islet Glucohinase mRNA" “Prevention and treatment of NIDDM"Editol by Y.Goto,et al,SmithーGordon,London,1992. (1992)

S. Hinata、S. Nishi 等人：“大鼠胰岛葡萄糖苷酶 mRNA 的激素调节”“NIDDM 的预防和治疗”Editol，Y. Goto 等人，Smith-Gordon，伦敦，1992 年。(1992)

西 重生,他: "膵ラ氏島細胞Glucokinaseの遺伝子発現調節" 糖尿病記録号1992 第35回日本糖尿病学会総会記録. (1992)

Shigeo Nishi等人：“胰岛细胞葡萄糖激酶的基因表达的调节”糖尿病记录第1992号日本糖尿病学会第35次总会记录(1992年)。

S.Hinata,S.Nishi,et al.: "Hormonal Regulation of Rat Pancreatic Islet Glucokinase messenger RNA" Excepta Medica.

S.Hinata、S.Nishi 等人：“大鼠胰岛葡萄糖激酶信使 RNA 的激素调节”Exceta Medica。