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目的探讨热应激对小鼠胸腺细胞凋亡及细胞周期情况的影响。方法 1月龄雄性小鼠随机分为6组(A~F),每组5只,A为对照组,B~F为实验组。对照组不进行热应激处理,实验组热应激(37℃)处理2h后恢复室温饲养,于恢复室温后0、2、6、10、20h依次取每组小鼠的胸腺。透射电子显微镜观察细胞形态的变化;流式细胞术、苏木素-伊红(HE)染色分析热应激后不同时间点小鼠胸腺细胞的凋亡及细胞周期情况;免疫组织化学染色检测各时间点小鼠胸腺细胞增殖细胞核抗原(PCNA)和热休克蛋白70(HSP70)的表达情况。结果热应激后2~10h电镜观察均可见典型的凋亡细胞形态;热应激后6h,胸腺细胞的凋亡率及S期细胞数量达到最高,而G0/G1、G2/M期细胞均降至最低,此时间点PCNA表达量较高。HSP70在热应激结束后0h即开始表达,2~6h表达量最高。结论热应激能够诱导小鼠胸腺细胞的凋亡,而HSP70则在凋亡过程中发挥保护作用。
Objective To investigate the effect of heat stress on thymocyte apoptosis and cell cycle in mice. Methods One-month-old male mice were randomly divided into 6 groups (A ~ F) with 5 rats in each group. A was the control group and B ~ F was the experimental group. The control group was not subjected to heat stress. The experimental group was treated with heat stress (37 ℃) for 2 hours and then reverted to room temperature. Thymus of each group was taken at 0, 2, 6, 10 and 20 hours after the recovery of room temperature. The changes of cell morphology were observed by transmission electron microscopy. The apoptosis and cell cycle of thymocytes were detected by flow cytometry and hematoxylin-eosin (HE) staining at different time points. Immunohistochemical staining was used to detect the changes of cell morphology at each time point Expression of proliferating cell nuclear antigen (PCNA) and heat shock protein 70 (HSP70) in mouse thymocytes. Results The morphological changes of apoptotic cells were observed after 2 ~ 10 h of heat stress. The apoptosis rate of thymocytes and the number of S phase cells reached the maximum at 6 h after heat stress, while the cells in G0 / G1 and G2 / M phases To a minimum, this time point higher PCNA expression. HSP70 began to express at 0h after the end of heat stress, and reached the peak at 2 ~ 6h. Conclusion Heat stress can induce apoptosis in mouse thymocytes, while HSP70 plays a protective role in apoptosis.