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目的观察不同脉宽5mW/cm2微波辐照对人AHH-1 T淋巴细胞的影响,探讨低剂量微波诱发的免疫细胞损伤机制及微波防护安全阈值。方法人AHH-1 T细胞经5mW/cm2模拟微波源辐射,根据不同脉宽(0、100、500、2000ns)分为对照(0ns)组、100、500、2000ns组。采用麦格-姬姆萨(MGG)染色和联脒-2-苯基吲哚(DAPI)染色法观察细胞凋亡形态,AnnexinV-FITC流式细胞仪检测细胞凋亡率和坏死率,MTT法检测细胞增殖活性,并用流式细胞仪检测细胞内活性氧自由基(ROS)水平和线粒体跨膜电位。结果 AHH-1 T细胞经2000ns脉宽的微波辐照后,细胞膜完整且核染色质正常,未见明显的凋亡细胞形态,细胞凋亡率和坏死率很低。经100ns脉宽的微波辐照后6、12、24h细胞增殖活性受抑,分别为对照组的95%、71%、74%,而2000ns组辐照后6、24h细胞增殖活性略有升高。在不同脉宽条件下,5mW/cm2微波辐照后细胞内ROS水平略呈下降趋势,且随脉宽升高而降低。100ns微波辐照后1h细胞跨膜电位略有下降,而500、2000ns组略有升高。结论 T淋巴细胞经不同脉宽5mW/cm2微波辐照后,细胞凋亡率和坏死率很低,细胞增殖活性轻度受抑,ROS水平和跨膜电位变化不大,未观察到明显的损伤效应,因此可作为避免微波辐照导致机体免疫损伤的安全阈值。
Objective To observe the effects of microwave irradiation with different pulse widths of 5 mW / cm2 on human AHH-1 T lymphocytes and to explore the mechanism of immune cell injury induced by low dose microwave and the safety threshold of microwave protection. Methods Human AHH-1 T cells were irradiated by 5 mW / cm2 simulated microwave source and divided into control (0ns), 100, 500 and 2000ns groups according to different pulse widths (0, 100, 500, 2000ns) The morphology of apoptotic cells was observed by using MGG staining and DAPI staining. The apoptosis rate and necrosis rate were detected by Annexin V-FITC flow cytometry. MTT assay The cell proliferation activity was measured. The level of reactive oxygen species (ROS) and mitochondrial transmembrane potential were measured by flow cytometry. Results After irradiated with 2000 ns pulse width, the AHH-1 T cells had intact cell membrane and normal chromatin. There was no obvious morphological changes of apoptotic cells. The apoptotic rates and necrosis rates of AHH-1 T cells were very low. After 100ns pulse microwave irradiation, the cell proliferative activity was inhibited at 6, 12 and 24 hours, respectively, 95%, 71% and 74% of the control group, while the cell proliferative activity increased slightly at 6 and 24 hours after irradiation . Under different pulse widths, the intracellular ROS level decreased slightly after 5mW / cm2 microwave irradiation, and decreased with the increase of pulse width. The transmembrane potential decreased slightly 1 h after 100 ns microwave irradiation, but slightly increased at 500 and 2000 ns. Conclusions After T lymphocytes were irradiated by 5mW / cm2 of different pulse widths, the apoptotic rate and necrosis rate were low, the cell proliferative activity was mildly inhibited, the level of ROS and transmembrane potential changed little, and no obvious damage was observed Effect, and therefore can be used as a safety threshold to avoid immune damage caused by microwave radiation.