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目的：探讨重组微小腺病毒载体（miniAd-ATP7B-GFP）对高铜孵育下的Wilson 病患者皮肤成纤维细胞铜代谢的影响。方法构建含人ATP7B基因的微小腺病毒载体；检测8例Wilson病患者突变基因；原代培养8例Wilson病患者与8名健康对照的皮肤成纤维细胞，使用基本培养液及铜浓度为22.3（C1）、89.2（C2）、156.1（C3）、245.3μmol／L（C4）培养液分别孵育2组细胞72 h，检测铜与蛋白含量。向Wilson病患者成纤维细胞分别转染miniAd-ATP7B-GFP（miniAd-ATP7B-GFP组）与空载体（ miniAd-GFP组），设立Wilson病未转染组及健康组做对照，用C4浓度培养液孵育4组细胞72、96 h，后检测各组细胞铜与蛋白含量。结果8例Wilson病患者检出5种突变基因，基本培养液及C1～C3组Wilson病患者与健康人铜／蛋白比值差异无统计学意义，C4组Wilson病患者[（1871.6±209.2） ng／mg]与健康人[（1267.2±188.3） ng／mg]铜／蛋白比值差异有统计学意义（ t＝6.075，P＜0.01），C3[（816.3±113.9） ng／mg]、C4组较基本培养液组[（159.2±38.6） ng／mg]差异有统计学意义（患者组χ2＝31.493，健康组χ2＝30.708，均P＜0.01）。96 h各组铜／蛋白比值均高于72 h组，96 h [（2071.0±171.8） ng／mg]、72 h[（1495.5±161.4） ng／mg]的miniAd-ATP7B-GFP组铜／蛋白比值与Wilson病未转染组[96 h（2731.2±188.7） ng／mg、72 h（1901.7±219.5） ng／mg]相比差异有统计学意义（72 h组F＝20.130，96 h 组F＝51.496，P＜0.01），与健康组比差异亦有统计学意义。结论MiniAd-ATP7B-GFP对高铜孵育下的Wilson病患者皮肤成纤维细胞的铜代谢有部分改善作用。

Objective To explore the effect of miniAd-ATP7B-GFP on the copper metabolism of skin fibroblasts of Wilson′s disease ( WD ) patients under high concentration copper medium.Methods Firstly, mini-adenovirus carrier containing human ATP7B gene was built and the mutations of 8 WD patients were detected.Fibroblasts from primary culture of skin of WD patients and normal human were cultivated 72 h in basic medium and medium with the copper concentration of C1(22.3μmol/L), C2(89.2μmol/L), C3 (156.1 μmol/L), C4 (245.3 μmol/L).Then the concentration of copper and protein was detected and copper/protein ratio was calculated.Secondly, miniAd-GFP ( miniAd-GFP group) and miniAd-ATP7B-GFP ( miniAd-ATP7B-GFP group ) were added into WD patients skin fibroblasts respectively, Wilson non-transfection group and normal group were set up as control, and C4 medium was used to culture the cells of four groups for 72 h and 96 h.Then the concentration of copper and protein was detected and copper/protein ratio was calculated.Results Five kinds of mutations were detected from 8 WD patients.The copper/protein ratio of WD patients and normal human in basic medium and the C1 -C3 groups had no statistically significant difference, but in C4 group (WD (1 871.6 ±209.2) ng/mg, normal group (1 267.2 ±188.3) ng/mg) the difference was statistically significant (t=6.075, P<0.01).C3((816.3 ±113.9) ng/mg) and C4 groups had statistically significant difference compared with the basic medium group ( ( 159.2 ± 38.6) ng/mg;WD:χ2 =31.493, normal group:χ2 =30.708, both P<0.01).The copper/protein ratio of 96 h group was higher than 72 h group.Compared with WD non-transfection (96 h:(2 731.2 ±188.7) ng/mg,72 h:(1 901.7 ±219.5) ng/mg) and normal groups, miniAd-ATP7B-GFP group had statistically&nbsp;significant difference both in 96 h ( ( 2 071.0 ±171.8 ) ng/mg ) and 72 h groups ( ( 1 495.5 ±161.4 ) ng/mg;72 h:F=20.130, 96 h: F=51.496,P<0.01).Conclusion MiniAd-ATP7B-GFP has partial improvement on copper metabolism of skin fibroblasts of WD patients under high concentration copper medium.