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目的观察亚砷酸钠(i As3+)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。
Objective To observe the effect of sodium arsenite (i As3 +) on the skin protein disulfide isomerase (PDI) in rabbits and to explore the relationship between PDI and arsenic-induced skin lesions. Methods Thirty healthy male rabbits were randomly divided into control group (C) and low (L, 0.13 mg / kg body weight (kg.w)), medium 0.65mg / kg.w), high (H, 1.30mg / kg.w) dose exposure group. Sodium arsenite water poisoning after 3 months of sacrifice, take the same position on the back full thickness skin. Two-dimensional gel electrophoresis was used to separate the proteins in the samples. The Bio-Rad image system was used to analyze the gel and the differentially expressed proteins were screened. The proteins were digested in gel and identified by matrix-assisted laser desorption / ionization time-of-flight mass spectrometry. The PDI mRNA expression in the samples was detected by real-time polymerase chain reaction (RT-PCR). Results There was no death in rabbits after 3 months of exposure. The body weight of rabbits in each group showed no significant change. The dorsum and ventral hair of the rabbits in the M, MH and H groups were dull and rough, especially in the H group. PDI in the skin of M, MH group decreased expression in the H group expression increased. PDI mRNA was up-regulated in MH and H groups (F = 225.83, P <0.001). Conclusion After exposure to sodium arsenite, the expression of PDI protein in rabbits decreased first and then increased with the increase of exposure dose, and the PDI mRNA gradually increased. At the same time, the hairs of rabbits were observed to be rough, Injury play a role.