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表达的影响

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.

Effects of hyperglycemia on the Ca2+ oscillation of mouse oocytes and expression of tumor necrosis factor related apoptosis inducing ligand of granulosa cells

Objective To study the effects of hyperglycemia on CaSUP>2+/SUP> oscillation of mouse preovulatory oocytes and the expression of tumor necrosis factor-related apoptosis inducing ligand(TRAIL) in granulosa cells. Methods Twenty day 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). CaSUP>2+/SUP> 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 CaSUP>2+/SUP>oscillation frequency was observed in the oocytes from the hyperglycemia group at 2 hours and 6 hours intervals ( EM>P/EM> <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( EM>P /EM><0.05),respectively. Conclusion Hyperglycemia increased CaSUP>2+/SUP> oscillation frequency of mouse preovulatory oocytes; Hyperglycemia increased express