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HSP70在不同应激程度CUMS昆明小鼠胚胎中的表达及作用

Expression of HSP70 in Kunming Mouse Embryos Stimulated by Chronic Mild Anticipatory Stress

  • 摘要: 目的建立雌性昆明小鼠慢性轻度不可预见性应激(CUMS)动物模型,探讨在心理应激状态下,热休克蛋白70(HSP70)是否参与机体对胚胎的应激保护作用。方法将300只雌性昆明小鼠以9种应激因子在28 d内随机应用造模,并分为轻度、中度和重度应激3组。对照组(50只)不造模。通过孕马血清促性腺激素/人绒毛膜促性腺激素促排卵,观察卵巢反应及胚胎发育潜能;采用免疫荧光染色及实时荧光定量PCR检测HSP70在2-细胞胚胎及孕4 d(D4)胚胎中的表达。结果28 d CUMS刺激后,实验组小鼠轻度应激占50%(150只),中度应激占32%(96只),重度应激占18%(54只)。促排卵后,轻度应激组小鼠2-细胞胚胎数、D4胚胎数和优胚率等与对照组比较差异均无统计学意义,而中、重度应激组小鼠较对照降低(P<0.05)。免疫荧光染色和实时荧光定量PCR显示HSP70在轻、中度应激组小鼠2-细胞胚胎和D4胚胎中的表达高于对照组(P<0.05),而重度应激组较对照组差异无统计学意义。结论CUMS刺激后HSP70的表达升高,可能与其应激保护作用有关。

     

    Abstract: Objective To measure the expression of heat shock protein 70 (HSP70) in mice embryos with the stimulation of chronic mild anticipatory stress (CUMS) to female Kunming mice. Methods Three hundreds female Kunming mice were stressed by 9 chronic mild unpredictable stress factors for 28 days and then divided into three groups of mild, moderate and severe stress. PMSG/hCG was measured to assess the induction of superovulation, and ovarian response and embryo development potential were observed. The expression of HSP70 in 2-cell embryos and day 4 embryos was detected by immunofluorescence and real time polymerase chain reaction (RT-PCR). Results After 28 d CUMS stimulation, the rate of mice in mild, moderate and severe stress were 50%, 32% and 18%,respectively. In the mild stress group, ovarian response and oocyte development potential were similar to those of control, while HSP70 expression of the embryos was significantly higher (P<0.05). In the severe stress group, ovarian response and oocyte development potential were significantly decreased compared with the control group (P<0.05), while HSP70 expression was similar to that of control. Conclusion HSP70 overexpression observed in the embryos may be related to its proteetive effect against chronic unpredictable stress.

     

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