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目的探讨香烟烟雾提取物(CSE)对原代培养的皮肤成纤维细胞生长和增殖的影响。方法采用噻唑蓝(MTT)比色法检测体外皮肤成纤维细胞存活力的影响,观察染毒后细胞形态和结构变化。β-半乳糖苷酶染色方法检测染毒后多次传代后成纤维细胞β-半乳糖苷酶的变化情况。用分光光度法检测胞质内活性氧自由基(ROS)含量、超氧化物歧化酶(SOD)活性。结果染毒后细胞呈典型的损伤形态,随着CSE浓度的增大和作用时间的延长,细胞存活力下降。长期染毒的细胞经5次传代后,β-半乳糖苷酶呈显著阳性;生长曲线示CSE作用后的细胞生长速度明显减慢。CSE作用于细胞后,细胞质内ROS含量升高(P<0.05),SOD活性降低(P<0.05)。结论CSE对体外培养的皮肤成纤维细胞增殖和生长有一定的抑制作用。低浓度CSE长期作用,可以促使原代培养的成纤维细胞迅速衰老。
Objective To investigate the effects of cigarette smoke extract (CSE) on the growth and proliferation of primary cultured human dermal fibroblasts. Methods The effect of MTT assay on the viability of human dermal fibroblasts in vitro was observed. Morphological and structural changes of the cells were observed. The changes of β-galactosidase in fibroblasts after multiple passages were detected by β-galactosidase staining. The contents of reactive oxygen species (ROS) and superoxide dismutase (SOD) in the cytoplasm were detected by spectrophotometry. Results The cells showed typical damage morphology after exposure. With the increase of CSE concentration and the prolongation of action time, the cell viability decreased. Long-term exposure of cells after 5 passages, β-galactosidase was significantly positive; growth curve showed that CSE effect of cell growth significantly slowed down. After CSE was applied to cells, ROS content in the cytoplasm increased (P <0.05) and SOD activity decreased (P <0.05). Conclusion CSE can inhibit the proliferation and growth of dermal fibroblasts cultured in vitro. Long-term effects of low concentrations of CSE, can promote primary cultured fibroblasts rapidly aging.