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目的研究低剂量电离辐射诱导小鼠睾丸一氧化氮(NO)含量、一氧化氮合酶(NOS)活性及凋亡诱导因子(apoptosis inducing factor,AIF)基因的变化。方法酶法检测不同剂量(0、0.025、0.05、0.075、0.1及0.2Gy)及0.075Gy照射后不同时间(0、3、6、12、18和24h)小鼠睾丸组织中NO含量及NOS活性的变化;Real-time PCR和Western blot法检测不同剂量(0、0.05、0.075、0.1及0.2Gy)及0.075Gy照射后不同时间(0、6、12和24h)小鼠睾丸组织中AIF mRNA表达及蛋白的表达。结果不同剂量X射线照射后12h,小鼠睾丸组织中NO含量及NOS活性随照射剂量增加而增加,0.075Gy增至最大(P<0.05),0.1~0.2Gy一直维持较高水平(P<0.05);AIF mRNA随剂量增加而增加,0.1Gy时达到最大(P<0.05);而AIF蛋白在0.075Gy时达到最大。0.075Gy X射线照射后,NO含量与NOS活性随时间延长而增加,NO含量在24h时增到峰值(P<0.05),而NOS活性在12h时达到峰值(P<0.05);AIFmRNA随时间延长而增加,在24h时达到峰值(P<0.05);其蛋白表达在12h时达到峰值。结论低剂量电离辐射照射后,小鼠睾丸生精细胞中NO含量、NOS活性及AIF基因表达增加,具有时程和剂量效应规律性。
Objective To study the changes of testis nitric oxide (NO), nitric oxide synthase (NOS) activity and apoptosis inducing factor (AIF) gene in mice induced by low dose ionizing radiation. Methods The levels of NO and NOS in testes of mice at different doses (0,0.025,0.05,0.075,0.1,0.2Gy) and 0.075Gy irradiation (0,3,6,12,18 and 24h) Real-time PCR and Western blot were used to detect AIF mRNA expression in testis tissue of mice at different times (0, 0.05, 0.075, 0.1 and 0.2Gy) and 0.075Gy And protein expression. Results NO and NOS activity in testis tissue of mice increased with the increase of irradiation dose at 12h after X-ray irradiation at various doses, with a maximum of 0.075Gy (P <0.05) and a high level of 0.1-0.2Gy (P <0.05) ); AIF mRNA increased with dose, reaching the maximum at 0.1Gy (P <0.05); while AIF protein reached the maximum at 0.075Gy. After 0.075Gy X-ray irradiation, NO content and NOS activity increased with time, the content of NO increased to peak at 24h (P <0.05), NOS activity peaked at 12h (P <0.05) (P <0.05). The protein expression peaked at 12h. Conclusion After exposure to low doses of ionizing radiation, NO, NOS and AIF expression in spermatogenic cells of mouse testes increase with time and dose-dependent regularity.