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目的通过分析长波紫外线(UVA)和中波紫外线(UVB)对Ha Ca T细胞作用后DNA拉曼光谱,从分子水平探索两者对Ha Ca T细胞DNA损伤作用的差异。方法取对数生长期的Ha Ca T细胞随机分为对照组、UVA组和UVB组。对照组3个样品,不进行紫外辐照(相当于照射前);后2组分别于接受UVA和UVB照射(累计照射剂量分别为8.26×10-2和2.97×10-2J/cm2)后,培养至照后0.5、1.0、2.0、3.0、4.0和5.0 h时收获细胞提取DNA,每个时间点3个样品。采用激光共焦拉曼光谱倒置显微镜系统记录各组Ha Ca T细胞全基因组DNA的拉曼光谱,对特征谱线进行归属,观察照射前后拉曼光谱特征谱线相对强度(Ιn/Ι0)变化。结果鸟嘌呤氢键、磷酸基静电斥力、腺嘌呤碱基分子内能和腺嘌呤碱基堆积力特征频率分别是669、1 094、1 230和1 344 cm-1。UVB组鸟嘌呤氢键、磷酸基静电斥力、腺嘌呤碱基分子内能的特征频率Ιn/Ι0均小于UVA组(P<0.01);上述3个指标在紫外辐射类型和照后时间的交互效应上均有统计学意义(P<0.01),UVB的损伤作用峰值均出现在照后1.0 h。UVB组腺嘌呤碱基堆积力特征频率Ιn/Ι0大于UVA组(P<0.01);紫外辐射类型和观察时间的交互效应有统计学意义(P<0.01),UVB的损伤作用峰值出现在照后3.0 h。结论 UVB的损伤作用对Ha Ca T细胞DNA的鸟嘌呤氢键、磷酸基静电斥力、腺嘌呤碱基分子内能和腺嘌呤碱基堆积力的损伤作用比UVA大,UVB的损伤作用峰值出现在照后1.0~3.0 h。
OBJECTIVE: To explore the DNA damage effects of HaCa T cells and HaCa T cells by molecular Raman spectroscopy after analyzing DNA Raman spectra of HaCa T cells by UVA and UVB. Methods Logarithmic growth phase HaCa T cells were randomly divided into control group, UVA group and UVB group. The control group of three samples, without UV irradiation (equivalent to before irradiation); the latter two groups were irradiated with UVA and UVB (cumulative dose of 8.26 × 10-2 and 2.97 × 10-2J / cm2) Cells were harvested at 0.5, 1.0, 2.0, 3.0, 4.0 and 5.0 h after culture to obtain DNA, 3 samples at each time point. The laser Raman spectroscopy inverted microscope system was used to record the Raman spectra of whole genome DNA of HaCa T cells in each group. The characteristic spectra were assigned to observe the relative intensity (Ιn / Ι0) of Raman spectra before and after irradiation. Results The characteristic frequencies of guanine hydrogen bond, phosphate radical electrostatic repulsion, adenine base internal energy and adenine base stacking force were 669,1 094,1 230 and 1 344 cm-1, respectively. The characteristic frequencies of guanine hydrogen bond, phosphate radical electrostatic repulsion and adenine base intramolecular energy in UVB group were lower than those in UVA group (P <0.01). The interaction between the above three indexes in ultraviolet radiation type and time after irradiation (P <0.01). The peak value of UVB damage appeared at 1.0 h after irradiation. The characteristic frequency of accumulation of adenine bases in UVB group was higher than that in UVA group (P <0.01). The interaction between ultraviolet radiation type and observation time was statistically significant (P <0.01) 3.0 h. CONCLUSION: The damage of UVB to DNA of HaCa T cells is stronger than that of UVA in the damage of guanine hydrogen bond, phosphate radical electrostatic repulsion, adenine base intramolecular energy and adenine base buildup, and the peak of UVB damage appears in According to 1.0 ~ 3.0 h.