高血糖对小鼠卵母细胞Ca^(2+)振荡和卵泡颗粒细胞肿瘤坏死因子相关凋亡诱导配体表达的影响  

Effects of hyperglycemia on the Ca^(2+) oscillation of mouse oocytes and expression of tumor necrosis factor related apoptosis inducing ligand of granulosa cells

作　　者：邵翠萍[1] 陈睿[1] 吕丹瑜[1] 李英[1] 马万云[2] 

机构地区：[1]北京大学基础医学院人体解剖学与组织学胚胎学系,北京100191 [2]清华大学物理系原子分子纳米科学教育部重点实验室,北京100084

出　　处：《解剖学报》2011年第5期618-621,共4页Acta Anatomica Sinica

基　　金：国家自然科学基金重大研究计划"细胞编程和重编程的表观遗传机制"资助项目(90919012);北京大学基础医学院理科基地创新人才培养资助项目(89100-217755138)

摘　　要：目的探讨高血糖对小鼠卵母细胞Ca2+振荡和卵泡颗粒细胞肿瘤坏死因子相关凋亡诱导配体(TRAIL)表达的影响。方法出生后20d ICR雌鼠,用链脲酶素建立急性高血糖模型;高血糖模型雌鼠及正常雌鼠注射10IU/只孕马血清促性腺激素(PMSG)和人绒毛膜促性腺激素(hCG)超排卵,注射hCG后0h、2h、6h和10h取生发泡(GV)期卵母细胞,双光子激光扫描共焦显微镜检测GV期卵母细胞的Ca2+振荡;hCG注射后6h、10h取卵丘-卵母细胞复合体(COC),利用免疫荧光、Western blotting检测TRAIL的表达。结果 1.高血糖组卵母细胞Ca2+振荡频率高于正常对照组,于hCG注射后2h、6h差异显著(P<0.05);2.免疫荧光、Western blotting蛋白半定量检测显示,高血糖组颗粒细胞TRAIL表达量高于正常对照组,于hCG注射后6h、10h差异有显著性(P<0.05)。结论高血糖加快小鼠未成熟卵母细胞Ca2+振荡的频率;高血糖促进小鼠卵泡颗粒细胞TRAIL的表达。Objective To study the effects of hyperglycemia on Ca^2＋ oscillation of mouse preovulatory oocytes and the expression of tumor necrosis factor-related apoptosis inducing ligand（TRAIL） in granulosa cells. Methods Twentyday old female mice were used to make a hyperplycemia model with streptozocin. Both diabetic and normal age-matched control mice received superovulation treatment with 10IU of pregnant mares srum gonadatropin （PMSG） and human chorionic gonadotropin（hCG）. Ca^2＋ oscillation of germinal vesicle （GV）-stage oocytes collected after hCG injection （0 hours, 2 hours, 6 hours and 10 hours） were recorded with two-photon laser scanning microscopy （TPLSM）. The expression of TRAIL in cumulus oocyte complesx （COC） was detected by immunofluorescence and Western blotting at 6 hour and 10 hour after hCG injection. Results 1. Faster Ca^2＋ oscillation frequency was observed in the oocytes from the hyperglycemia group at 2 hours and 6 hours intervals （P 〈 0. 05） ; 2. Intensities for TRAIL positive immunofluorescence reactions and Western blotting staining intensities were stronger in the hyperglycemia groups than that in the control groups at the intervals of 6 hours and 10 hours（P 〈 0.05） ,respectively. Conclusion Hyperglycemia increased Ca^2＋ oscillation frequency of mouse preovulatory oocytes; Hyperglycemia increased expression of TRAIL of granulosa cells.

关 键 词：高血糖 卵母细胞 Ca^2+振荡 颗粒细胞 肿瘤坏死因子相关凋亡诱导配体 免疫荧光 免疫印迹法 小鼠 

分 类 号：Q344.5[生物学—遗传学]