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目的研究久效磷对体外培养的大鼠皮肤成纤维细胞凋亡及氧化损伤作用的影响。方法取4只3日龄清洁级SD大鼠的背部皮肤,以组织块法培养大鼠皮肤成纤维细胞,正常传代。取第4代成纤维细胞,调整细胞密度为1.0×106/瓶,当细胞至亚融合状态,分别加入0(对照)、0.01、0.1、1μg/L的久效磷溶液培养6、12、24 h。采用流式细胞术检测大鼠皮肤成纤维细胞凋亡率;采用分光光度比色法检测皮肤成纤维细胞超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活力和丙二醛(MDA)含量。结果 0.1～1μg/L久效磷染毒6、12、24 h时,大鼠皮肤成纤维细胞SOD、CAT活性均低于对照组(P<0.05或P<0.01),仅0.01μg/L久效磷染毒24 h时SOD活力低于对照组(P<0.05)。各浓度久效磷染毒大鼠皮肤成纤维细胞凋亡率和MDA含量均明显高于对照组(P<0.05或P<0.01)。随着久效磷染毒剂量的升高和染毒时间的延长,大鼠皮肤成纤维细胞SOD、CAT活力均呈下降趋势,而MDA含量和细胞凋亡率呈上升趋势。结论久效磷可诱导大鼠皮肤成纤维细胞氧化损伤和细胞凋亡率增高。
Objective To study the effect of monocrotophos on the apoptosis and oxidative damage of cultured rat skin fibroblasts in vitro. Methods Four dorsal skin of 3-day-old clean SD rats were used to culture rat skin fibroblasts by tissue block method and passaged normally. The 4th generation of fibroblasts were prepared, and the cell density was adjusted to 1.0 × 10 6 per flask. When the cells were sub-fused, 0, 0, 0.01, 0.1 and 1 μg / L monocrotophos solution h The apoptosis rate of rat skin fibroblasts was detected by flow cytometry. The activity of superoxide dismutase (SOD), catalase (CAT) and malondialdehyde (MDA) in skin fibroblasts were detected by spectrophotometry. )content. Results The activities of SOD and CAT in rat skin fibroblasts at 0.1, 1 μg / L monocrotophos exposure for 6, 12 and 24 h were lower than those in the control group (P <0.05 or P <0.01), and only 0.01 μg / L The activity of SOD was lower than that of the control group at 24 h (P <0.05). The apoptosis rate and MDA content of skin fibroblasts in each concentration of monocrotophos were significantly higher than those in the control group (P <0.05 or P <0.01). With the increase of the dose of monocrotophos and the prolongation of the exposure time, the activities of SOD and CAT in rat skin fibroblasts decreased, while the content of MDA and the rate of apoptosis increased. CONCLUSION Monocrotiplet-induced oxidative damage and apoptosis of rat skin fibroblasts.