高糖对小鼠精母细胞增殖和凋亡的影响

Effect of high glucose on proliferation and apoptosis of mouse spermatocyte

目的 探讨高糖对小鼠精母细胞增殖和凋亡的影响并初步探讨其机制。方法 体外培养小鼠精母细胞GC-2spd,分为正常对照组和高糖组,分别采用CKK-8法和流式细胞术检测细胞增殖、细胞周期和凋亡情况,实时荧光定量PCR和蛋白免疫印迹技术检测GC-2spd细胞caspase3、caspase9、Notch1、Jagged1和Hes1基因mRNA和蛋白的表达情况。结果 与正常对照组比较,高糖组GC-2spd细胞生长速度显著降低,G0/G1期细胞比例显著增多,早期、晚期凋亡率均显著增多,caspase3、caspase9mRNA及蛋白相对表达量显著上调(P均<0.05),Notch、Jagged1、Hes1mRNA及蛋白相对表达量显著下调(P<0.05)。结论 高糖可促进小鼠精母细胞凋亡,可能通过抑制Notch信号通路实现。

Objective : To investigate the effect of high glucose on the proliferation and apoptosis of mice spermatocyte and its mechanism. Methods : GC-2spd of mice spermatocytes were cultured in vitro and divided into normal control group and high glucose group. The cell proliferation was determined by CKK-8 assay,and the GC-2spd cells cycle and the apoptosis were detected by flow cytometry. The gene and protein expression of Notch1,Jagged1,Hes1,caspase3 and caspase9 in GC-2spd cells were detected by the real time fluorescent quantitative PCR (qRT-PCR) and Western blot(WB) respectively. Results : Compared with the control group,the growth rate of GC-2spd cells in the high glucose group was significantly reduced and the proportion of cells in G0/G1 phase was increased significantly. The apoptosis rate in early and late phases was significantly increased. The mRNA and protein expressions of Notch,Jagged1,Hes1 mRNAs were significantly decreased,but the mRNA and protein expressions of caspase3, caspase9 mRNAs were significantly up-regulated( P <0.05). Conclusions : High glucose can promote spermatocyte apoptosis in mice,which may be achieved by suppressing the Notch signaling pathway.