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目的 观察致死剂量 γ射线照射后小鼠脾淋巴细胞凋亡特征及其与 Bax和 Bcl XL 表达的关系。方法 清洁级 C5 7小鼠 2 2 5只 ,随机分为 0、6、9、12、15和 2 0 Gy6个组 ,经 γ射线全身一次照射 ,于照射前和照射后 6 h～ 2 8d活杀取材 ,用原位末端标记法 (TU NEL)和 Annexin V流式细胞术 (FCM)检测淋巴细胞凋亡和坏死情况 ,用碱性磷酸酶免疫组化技术检测 Bax和 Bcl XL 蛋白的表达。结果 1各剂量组小鼠外周血白细胞在照射后 6 h出现一过性升高 ,以后迅速下降。 6 Gy照射后 7d降至最低 ,至照射后 2 8d基本恢复到正常水平。 2不同剂量照射后 6 h脾淋巴细胞凋亡率达高峰 ;照射后 6～ 2 4 h,≤ 12 Gy照射后凋亡率随照射剂量的增加而升高 ,≥ 15 Gy照射后凋亡率下降。 3FCM检测结果证实 ,6～ 12 Gy照射后 6 h,脾淋巴细胞凋亡率呈现出量效关系 ,≥ 15 Gy照射后凋亡率下降 ,而坏死率明显升高。DNA凝胶电泳图谱分析支持上述结果。 4照射后 6 h,6 Gy照射后脾淋巴细胞 Bax蛋白表达增加 ,至 12 Gy达峰值 ,显示出量效关系 ;≥ 15 Gy未显示出量效关系 ;Bcl XL 蛋白随照射剂量增加而持续降低 ,在 6～ 12 Gy也显示出较好的量效关系。结论 6～ 12 Gy照射后脾淋巴细胞死亡方式以凋亡为主 ,≥ 15 Gy照射后凋亡与坏死均为
Objective To observe the apoptosis characteristics of splenic lymphocytes and its relationship with the expression of Bax and Bcl-XL in lethal dose γ-rays in mice. Methods Twenty-five clean mice of C5 7 were randomly divided into 6 groups of 0, 6, 9, 12, 15 and 20 Gy. The whole body was irradiated with γ-rays. The animals were irradiated before and 6 h to 28 d after irradiation The apoptosis and necrosis of lymphocytes were detected by TU NEL and Annexin V flow cytometry (FCM). The expressions of Bax and Bcl XL were detected by alkaline phosphatase immunohistochemistry. Results 1 The peripheral blood leukocytes of mice in each dose group showed a transient increase 6 h after irradiation, and then decreased rapidly thereafter. 6 Gy 7d after irradiation to a minimum, to 28d after irradiation basically returned to normal levels. The apoptosis rate of splenic lymphocytes reached the peak at 6 h after irradiation at different doses. The apoptotic rate of spleen lymphocytes at 6 ~ 24 h and 12 Gy after irradiation increased with the increase of irradiation dose, and the apoptosis rate decreased after 15 Gy irradiation . 3FCM test results showed that 6 ~ 12 Gy 6 h after irradiation, the spleen lymphocyte apoptosis rate showed a dose-response relationship, 15 Gy irradiation apoptosis rate decreased, and the necrosis rate was significantly increased. DNA gel electrophoresis analysis supports the above results. 4 at 6 h, the expression of Bax protein in spleen lymphocytes increased after 6 Gy irradiation, reaching the peak value at 12 Gy, showing a dose-response relationship; ≥ 15 Gy showed no dose-response relationship; and the Bcl XL protein continuously decreased with increasing radiation dose , 6 ~ 12 Gy also showed a good dose-response relationship. Conclusion The apoptosis of splenic lymphocytes is predominated by 6 ~ 12 Gy irradiation. The apoptosis and necrosis after irradiation of ≥ 15 Gy are