论文部分内容阅读
目的 探讨脑源性神经营养因子(BDNF)对正常和热应激条件下小鼠着床前胚胎发育和凋亡的影响.方法 150只雌性昆明小鼠用于实验,2细胞期胚胎添加或不添加10μg/L BDNF进行体外培养,早期囊胚37℃条件下或41℃热处理2h,观察胚胎发育情况,TUNEL法和Hoechst 33342分析囊胚细胞数量和凋亡情况,荧光技术分析Caspase-9的活力.结果 在37℃条件下BDNF能够促进早期囊胚发育,增加囊胚细胞数量,降低凋亡率及Caspase-9的活力,热应激能够降低早期囊胚的发育能力,减少囊胚细胞数量而增加凋亡率,且中等和高Caspase-9活力囊胚的比例热处理后明显增加,当培养基中添加10μg/L BDNF时,能够明显缓解热应激对早期囊胚发育和质量的不利影响.结论 BDNF对37℃或41℃热应激条件下小鼠着床前胚胎发育、细胞数量、凋亡和Caspase-9活力起到一定的调控作用.“,”Objective To investigate the effect of brain derived neurotrophic factor (BDNF) on the development and apoptosis of mouse early blastocyst either cultured continuously at 37 ℃ or treated with heat shock at 41 ℃ for 2 hours.Methods A total of 150 female Kunming mice were used.The early blastocysts were cultured continuously at 37 ℃ or subjected to heat shock for 2 hours at 41 ℃ while being cultured with or without 10μg/L BDNF.The development of early blastocysts was recorded.The TUNEL reaction and Hoechst 33342 method were used to observe the apoptosis cells of blastocysts.The Caspase-9 activity of mouse blastocyst was analysed by fluorescent technique.Results Treatment with BDNF at 37 ℃ promoted the development of early blastocysts.The normal developmental blastocyst cells were increased while apoptosis cells and Caspase-9 activities were decreased in early blastocyst in BDNF treatment group.The development of early blastocyst was inhibited in heat shock treatment group,and TUNEL positive cells number and Caspase-9 activities were increased.There were normal developmental blastocyst in 10μg/L BDNF treatment group.Conclusion Our findings suggest that BDNF signaling plays important roles during mouse early blastocyst development,blastocyst cell numbers,blastocyst apoptosis and the Caspase-9 activity.