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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 investigate the effects of hyperglycemia on the oocyte calcium oscillations and the expression of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) in mouse granulosa cells. METHODS: ICR female rats were induced 20 days after birth with streptozotocin to establish an acute hyperglycemic model. Hyperglycemia female and normal female mice were injected with 10IU / pregnant horse serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG) Ovulation, Ovulation of GV oocytes at 0h, 2h, 6h and 10h after injection of hCG, Ca2 + oscillations in GV oocytes were detected by two-photon laser scanning confocal microscope. At 6h and 10h after hCG injection, - oocyte complex (COC), the use of immunofluorescence, Western blotting detection of TRAIL expression. The results showed that the frequency of Ca2 + oscillations in hyperglycemia group was higher than that in normal control group at 2h and 6h after hCG injection (P <0.05) .2. Immunofluorescence and Western blotting showed that the frequency of Ca2 + The expression of TRAIL in the cells was higher than that in the normal control group, and the difference was significant at 6h and 10h after hCG injection (P <0.05). Conclusion Hyperglycemia accelerates the frequency of Ca2 + oscillations in mouse immature oocytes. Hyperglycemia promotes the expression of TRAIL in mouse granulosa cells.