Expression change of protein disulfide isomerase in skin tissues of rabbits exposed to arsenic
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摘要: 目的 观察亚砷酸钠(iAs3+)染毒对家兔皮肤蛋白质二硫键异构酶(protein disulfide isomerase,PDI)的影响,探讨PDI与砷致皮肤损害的关系。方法 将30只健康雄性家兔随机平均分到对照组(C)及低(L,0.13mg/千克体重(kg·w))、中(M,0.26mg/kg·w)、中高(MH,0.65mg/kg·w)、高(H,1.30mg/kg·w)剂量染毒组。亚砷酸钠饮水染毒3个月后处死,取背部相同位置全厚皮肤样。双向凝胶电泳分离样品中蛋白,Bio-Rad图像系统分析胶图,筛选差异表达蛋白,胶内酶切后采用基质辅助激光解析电离飞行时间质谱进行鉴定。通过实时荧光定量聚合酶链反应(real-time polymerase chain reaction,RT-PCR)检测样品中PDI mRNA的表达。结果 染毒3个月后,家兔无死亡,各组家兔体重无明显变化,M、MH、H组可观察到家兔背腹部毛发无光泽、粗糙,H组尤为明显。皮肤中PDI在M、MH组表达量下降,在H组表达量上升。PDI mRNA在MH和H组上调(F=225.83,P<0.001)。结论 亚砷酸钠染毒后,随染毒剂量升高,家兔皮肤PDI蛋白表达量先降低后升高, PDI mRNA逐渐增高,同时可观察到家兔毛发粗糙,提示PDI可能在砷致皮肤损伤中发挥作用。Abstract: Objective To explore the influence of exposure to sodium arsenite(iAs3+) on protein disulfide isomerase (PDI) in skin tissues of rabbits and discuss the relationship between arsenic poisoning on skin and PDI. Methods A total of 30 healthy male rabbits was divided into 5 groups including contrast (C) group, low-dose group (L, 0.13 mg/kg·w), medium-dose group (M, 0.26mg/kg·w), medium-high-dose group (MH, 0.65mg/kg·w), and high-dose group (H, 1.30 mg/kg·w) treated with the above different doses of sodium arsenite, 6 rabbits for each group. After free drink water had been poisoned for consecutive 3 months, these animals were killed and the skin tissues on the same place of these rabbits' back were taken for sampling. Differential expression proteins between arsenite poisoning groups and contract group were screened using two-dimensional gel electrophoresis (2-DE) and identified by MALDI-TOF-MS. The expression level of PDI mRNA of skin samples of rabbitss were tested through Real-Time Quantitative PCR. Results After being poisoned for 3 months, no rabbit died, coarse fur of rabbits' back and belly were observed in M, MH, and H groups, and this condition was especially obvious in H group. It was discovered that the expression quantity of PDI was reduced in M and MH groups while was increased in H group. The expression level of PDI mRNA was increased in MH and H groups (F=225.83, P<0.001). Conclusions Sodium arsenite poisoning can change PDI mRNA and the protein expression quantity in skin tissue of rabbits. With the increasing of exposure doses, PDI mRNA expression was increased, and meanwhile coarse fur of rabbits was discovered. However, PDI protein expression was first decreased and then increased. This suggests that PDI might play a role in that arsenite damages skin.
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