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目的探讨虾青素对辛硫磷所致大鼠皮肤成纤维细胞氧化损伤及凋亡的拮抗作用。方法取4只3日龄清洁级SD大鼠背部皮肤培养。取第4代成纤维细胞,正常对照组和辛硫磷氧化损伤组更换新鲜培养基,低、中、高剂量虾青素保护组分别更换含0.2、2、20μg/L虾青素的培养基,于37℃、5%CO2培养24 h后,正常对照组更换新鲜培养基,辛硫磷氧化损伤组和各剂量虾青素保护组分别更换含10μg/L辛硫磷的培养基,于37℃、5%CO2培养24 h。采用噻唑兰(MTT)比色法检测细胞存活率;采用流式细胞术检测细胞凋亡率;测定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。结果与对照组比较,辛硫磷染毒组和虾青素保护组大鼠皮肤成纤维细胞存活率和SOD、CAT活力下降,凋亡率和MDA含量升高。与辛硫磷染毒组比较,虾青素保护组大鼠皮肤成纤维细胞存活率和SOD、CAT活力较高,凋亡率和MDA含量下降;且随着虾青素剂量的升高,辛硫磷染毒大鼠皮肤成纤维细胞存活率和SOD、CAT活力呈上升趋势,凋亡率和MDA含量呈下降趋势。结论虾青素对辛硫磷所致的大鼠皮肤成纤维细胞氧化损伤及凋亡具有明显的拮抗作用。
Objective To investigate the antagonistic effect of astaxanthin on oxidative damage and apoptosis of rat skin fibroblasts induced by phoxim. Methods Four 3-day-old SD rats were cultured on the back of the skin. Take 4th generation of fibroblasts, normal control group and phoxim oxidative injury group to replace the fresh medium, low, medium and high doses of astaxanthin protection group were replaced with 0.2,2,20μg / L astaxanthin medium After incubating at 37 ℃ and 5% CO2 for 24 h, the normal control group was replaced with fresh medium, the phoxim oxidative injury group and each dose of astaxanthin protective group were replaced with 10μg / L phoxim in 37 ℃, 5% CO2 for 24 h. Cell viability was determined by MTT assay. Flow cytometry was used to detect the apoptosis rate. The activities of superoxide dismutase (SOD), catalase (CAT) and malondialdehyde (MDA) )content. Results Compared with the control group, the survival rate of the fibroblasts and the activities of SOD and CAT, the apoptotic rate and the content of MDA in the phoxim-treated and astaxanthin-treated groups were decreased. Compared with the phoxim group, the survival rate of skin fibroblasts and the activity of SOD and CAT in astaxanthin-protected group were higher, while the apoptotic rate and MDA content were decreased. With the increase of astaxanthin dose, Thioflavin-induced rat skin fibroblast survival rate and SOD, CAT activity showed an upward trend, the apoptotic rate and MDA content decreased. Conclusion Astaxanthin has a significant antagonistic effect on oxidative damage and apoptosis of rat skin fibroblasts induced by phoxim.