中华妇产科杂志
中華婦產科雜誌
중화부산과잡지
CHINESE JOUNAL OF OBSTETRICS AND GYNECOLOGY
2008年
1期
54-58
,共5页
闫妙娥%吴效科%宋娟娟%侯丽辉
閆妙娥%吳效科%宋娟娟%侯麗輝
염묘아%오효과%송연연%후려휘
胰岛素抗药性%1-磷脂酰肌醇3-激酶%雄二烯类%单糖转运蛋白质类%丝裂原激活蛋白激酶类
胰島素抗藥性%1-燐脂酰肌醇3-激酶%雄二烯類%單糖轉運蛋白質類%絲裂原激活蛋白激酶類
이도소항약성%1-린지선기순3-격매%웅이희류%단당전운단백질류%사렬원격활단백격매류
Insulin resistance%1-phosphatidylinositol 3-kinase%Androstadienes%Monosaccharide transport proteins%Mitogen-activated protein kinases
目的 观察磷脂酰肌醇3激酶(PI-3K)抑制剂--沃曼青霉素(wortmannin)诱导猪卵巢颗粒细胞胰岛素抵抗(IR)后,其信号传导通路中葡萄糖转运蛋白4(GLUT4)、丝裂原激活蛋白激酶(MAPK)基因表达的变化.方法 采用猪卵巢颗粒细胞体外培养,分别以浓度为0、1.0、1.5、3.0、5.0μmol/L的wortmannin作用48 h后,以氚标记葡萄糖法及葡萄糖氧化酶法检测细胞的葡萄糖消耗量,以免疫荧光法分析GLUT4、MAPK蛋白定位表达情况,以RT-PCR方法 对GLUT4、MAPK的mRNA进行半定量分析.结果 浓度为1.5 μmol/L的wortmannin降低细胞对葡萄糖的摄取量最高达40%(P＜0.05),残余葡萄糖含量增多约13.76%,出现明显的IR.免疫荧光法结果 显示GLUF4、MAPK蛋白主要在细胞质表达.RT-PCR结果 提示,GLUT4基因表达水平降低21.06%,MAPK表达水平升高56.43%.结论 wortmannin可干扰细胞内的糖代谢信号传导通路,诱导IR的发生;同时可放大有丝分裂信号传导通路.
目的 觀察燐脂酰肌醇3激酶(PI-3K)抑製劑--沃曼青黴素(wortmannin)誘導豬卵巢顆粒細胞胰島素牴抗(IR)後,其信號傳導通路中葡萄糖轉運蛋白4(GLUT4)、絲裂原激活蛋白激酶(MAPK)基因錶達的變化.方法 採用豬卵巢顆粒細胞體外培養,分彆以濃度為0、1.0、1.5、3.0、5.0μmol/L的wortmannin作用48 h後,以氚標記葡萄糖法及葡萄糖氧化酶法檢測細胞的葡萄糖消耗量,以免疫熒光法分析GLUT4、MAPK蛋白定位錶達情況,以RT-PCR方法 對GLUT4、MAPK的mRNA進行半定量分析.結果 濃度為1.5 μmol/L的wortmannin降低細胞對葡萄糖的攝取量最高達40%(P＜0.05),殘餘葡萄糖含量增多約13.76%,齣現明顯的IR.免疫熒光法結果 顯示GLUF4、MAPK蛋白主要在細胞質錶達.RT-PCR結果 提示,GLUT4基因錶達水平降低21.06%,MAPK錶達水平升高56.43%.結論 wortmannin可榦擾細胞內的糖代謝信號傳導通路,誘導IR的髮生;同時可放大有絲分裂信號傳導通路.
목적 관찰린지선기순3격매(PI-3K)억제제--옥만청매소(wortmannin)유도저란소과립세포이도소저항(IR)후,기신호전도통로중포도당전운단백4(GLUT4)、사렬원격활단백격매(MAPK)기인표체적변화.방법 채용저란소과립세포체외배양,분별이농도위0、1.0、1.5、3.0、5.0μmol/L적wortmannin작용48 h후,이천표기포도당법급포도당양화매법검측세포적포도당소모량,이면역형광법분석GLUT4、MAPK단백정위표체정황,이RT-PCR방법 대GLUT4、MAPK적mRNA진행반정량분석.결과 농도위1.5 μmol/L적wortmannin강저세포대포도당적섭취량최고체40%(P＜0.05),잔여포도당함량증다약13.76%,출현명현적IR.면역형광법결과 현시GLUF4、MAPK단백주요재세포질표체.RT-PCR결과 제시,GLUT4기인표체수평강저21.06%,MAPK표체수평승고56.43%.결론 wortmannin가간우세포내적당대사신호전도통로,유도IR적발생;동시가방대유사분렬신호전도통로.
Objective To investigate the biological effects of insulin resistance(IR)on the porcine granulosa cells which iS induced by wortmannin,the PI-3K inhibitor and mediated by key molecules including GLUT4 and MAPK during insulin signaling.Methods The model of IR porcine granulosa cell was established in in vitro culture by treatment of wortmannin,and was assessed the amount of3H glucose uptake as well as medium glucose levels by glucose oxidase method.The protein and mRNA expression of GLUT4 and MAPK were evaluated by immunofluorescence and RT-PCR respectively.Resuits The glucose intake was decreased by 40% with treatment of wortmannin at 1.5 μmol/L(P＜0.05).GLUT4 and MAPK were localized mainly to cytoplasm of grantdose cells.When granulosa cells were insulin resistant,the expression of GLUT4 was down-regulated whereas MAPK was up-regulated as compared with the controls.Conclusions Wortmannin treatment can lcad to decreased expression of GLUT4 and increase of IR granulose cells.This metabolic phenotype could induce increased expression of MAPK and mitogenic potential,indicating the cross-talk between two pathways of insulin signaling within ovarian cells.
