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目的建立长波紫外线(UVA)诱导的成纤维细胞急性光损伤模型。方法原代培养人皮肤成纤维细胞,采用形态学观察、免疫荧光细胞染色对皮肤成纤维细胞进行鉴定,分别用形态学观察、CCK-8法、细胞衰老β-半乳糖苷酶染色检测UVA对细胞形态、增生活性以及细胞衰老状态的影响。结果 UVA剂量≤4 J/cm2时对细胞形态无明显影响,而UVA剂量≥6 J/cm2时细胞出现变形;UVA剂量≤4 J/cm2时,细胞增生率均>85%,而UVA剂量≥6 J/cm2时细胞增生率明显下降,与对照组相比差异有统计学意义(P<0.01);4 J/cm2组的衰老阳性细胞明显增加,与对照组相比,差异有统计学意义(P<0.01)。结论 4 J/cm2 UVA可用于建立人皮肤成纤维细胞急性光损伤模型。
OBJECTIVE To establish a long-wave ultraviolet (UVA) induced acute injury model of fibroblasts. Methods Human dermal fibroblasts were cultured in primary culture. Skin fibroblasts were identified by morphological observation and immunofluorescence staining. Morphological observation, CCK-8 assay and β-galactosidase staining were used to detect UVA Cell morphology, proliferative activity and cell aging status. Results When the dosage of UVA was less than 4 J / cm2, the morphological changes of cells were not obvious. When UVA dose was> 6 J / cm2, the cells were distorted. When the dose of UVA was less than 4 J / cm2, the cell proliferation rate was> 85% The cell proliferation rate at 6 J / cm2 was significantly decreased compared with the control group (P <0.01). The number of senescence positive cells in 4 J / cm2 group was significantly increased compared with the control group, the difference was statistically significant (P <0.01). Conclusions 4 J / cm2 UVA can be used to establish a model of acute photodamage of human dermal fibroblasts.