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①目的探讨青紫薯色素对60Coγ-射线单次辐射小鼠胸腺淋巴细胞辐射损伤的保护作用及其体外抗氧化作用机制。②方法建立60Co辐射(强度为3 Gy)对体外培养的小鼠胸腺淋巴细胞损伤病理模型,分为6组:对照组6、0Co模型组、0.625 g/L青紫薯色素组、1.250 g/L青紫薯色素组、2.500 g/L青紫薯色素组6、0Co+1 g/L Vit C组。琼脂糖凝胶电泳观察DNA Ladder;流式细胞仪PI染色测定细胞凋亡率;Fluo-3-AM荧光染色测定细胞内游离Ca2+浓度;免疫细胞化学法测定P53蛋白的表达;原位杂交检测细胞p21 mRNA的表达;化学发光法和电子自旋共振(ESR)捕集技术检测青紫薯色素对超氧阴离子(O2-)、羟自由基(OH.)、过氧化亚硝基(ONOO-)和脂类自由基的清除作用。③结果0.625~2.500 g/L浓度范围内青紫薯色素能抑制60Co辐射所致淋巴细胞DNA梯状带的出现,降低淋巴细胞凋亡率,降低淋巴细胞内游离Ca2+浓度,抑制60Co辐射后小鼠胸腺淋巴细胞P53蛋白在胞浆的表达,抑制60Co辐射后小鼠胸腺淋巴细胞p21 mRNA的表达,同时青紫薯色素还可有效清除O2-、OH.、ONOO-和脂类自由基。④结论青紫薯色素能抑制60Co辐射诱导的淋巴细胞凋亡,对60Co辐射损伤的小鼠胸腺淋巴细胞具有保护作用。其作用机制与青紫薯色素能清除氧自由基、降低细胞内游离Ca2+浓度、减少DNA的断裂、抑制突变型P53蛋白的表达、抑制p21基因的表达,影响淋巴细胞凋亡的信号转导中的基因调控有关。
① Objective To investigate the protective effect of purple sweet potato pigment on the radiation injury of thymus lymphocytes in mice exposed to 60Coγ-ray and its anti-oxidative mechanism in vitro. Methods The pathological model of mouse thymus lymphocytes injury induced by 60Co radiation (intensity 3 Gy) was established and divided into 6 groups: control group 6,0Co model group, 0.625 g / L purple potato pigment group, 1.250 g / L Purple sweet potato pigment group, 2.500 g / L purple sweet potato pigment group 6,0Co + 1 g / L Vit C group. Agarose gel electrophoresis was used to observe DNA Ladder; flow cytometry PI staining was used to determine the apoptosis rate; Fluo-3-AM fluorescence staining was used to measure intracellular free Ca2 + concentration; immunocytochemistry was used to detect the expression of P53 protein; The expression of p21 mRNA was detected by chemiluminescence and electron spin resonance (ESR) techniques. The effects of purple pigment on superoxide anion (O2-), hydroxyl radical (OH), peroxynitrite (ONOO-) and Lipid free radical scavenging effect. ③ Results Potato purple pigment could inhibit the appearance of DNA ladder caused by 60Co radiation, decrease the rate of lymphocyte apoptosis, decrease the concentration of free Ca2 + in lymphocytes, and inhibit the radiation of 60Co radiation in the concentration range of 0.625 ~ 2.500 g / L The expression of P53 protein in the thymus lymphocytes and the expression of p21 mRNA in the thymus lymphocytes of mice after 60Co irradiation, while the purple potatoes pigment could effectively remove O2-, OH., ONOO- and lipid free radicals. ④ Conclusion Purple potato pigment can inhibit 60Co radiation-induced lymphocyte apoptosis, 60Co radiation-induced mouse thymus lymphocytes have a protective effect. Its mechanism of action and purple potato pigment can scavenge oxygen free radicals, reduce intracellular free Ca2 + concentration, reduce DNA breakage, inhibit the expression of mutant P53 protein, inhibit the expression of p21 gene and affect the signal transduction of lymphocyte apoptosis Gene regulation related.