MC148基因局部转染延长大鼠异体皮肤移植成活时间的研究

目的探讨MC148基因局部转染对皮肤移植排斥反应的免疫抑制作用。方法从传染性软疣病毒(MCV)中克隆MC148基因,构建Ad-MC148腺病毒重组体后,用其转染8只Lewis大鼠(供体)尾皮局部。另选16只Wistar大鼠(受体),于尾部制作皮肤缺损创面后,分为对照组8只, 移植未转染的供体尾皮;转染组8只,移植转染了MC148基因的供体尾皮。于术后6 h及2、3、7、10 d,检测MC148 mRNA在转染组大鼠皮肤组织内的表达情况,以它与内参照β肌动蛋白之间的吸光度 (A)之比表示;比较两组大鼠移植皮肤的成活时间;术后7 d起对两组部分大鼠行大体观察,其余大鼠于术后7 d行组织病理学观察。结果术后6 h转染组大鼠皮肤组织内有MC148 mRNA表达, 第3天达到高峰(A MC148 mRNA/Aβ肌动蛋白=0．86),随后下降,但可持续表达约10 d。对照组大鼠移植皮肤成活时间为(8．5±3．4)d,明显低于转染组(15．0±2．0)d(P<0．01)。大体及组织病理学观察到, 术后7 d转染组大鼠尾皮色泽红润,皮下组织内有少量炎性细胞浸润,12-20 d后移植皮肤才逐渐变黑、脱落;术后7 d对照组大鼠尾皮色泽变黑、逐渐脱落,真皮及皮下组织有大量炎性细胞浸润。结论 MC148基因局部转染可能通过抑制器官移植后白细胞浸润而发挥免疫抑制作用。

Prolongation of survival time of allogenous skin graft after transfection of Molluscum contagiosum virus 148 gene in rats

OBJECTIVE: To explore the immunosuppressive effect of local transfection of Molluscum contagiosum virus 148 (MC148) gene to allogenous skin graft against rejection. METHODS: MC148 gene was cloned from molluscum contagiosum virus (MCV), and was employed to construct recombinant adenovirus vector (Ad-MC148). The recombinant Ad-MC148 was then locally transfected into a part of the tail skin of eight Lewis rats, which served as skin donors for grafting. The wounds (1 cm x 1 cm) were produced on the tails of 16 Wistar rats, and they were then randomly divided into control (C, n=8, with grafting of skin from donor rats without transfection), and transfection (T, n=8, with grafting of skin from donor rats with transfection of the recombinant Ad-MC148) groups. The expression of MC148 mRNA gene in T group was detected on 6 post operation hour( POH) and 2, 3, 7 and 10 post operation day (POD), and the results were expressed by the ratio of absorption value (A) between MC148 gene and beta-actin. The survival time of skin grafts in both groups was compared. Gross examination of grafted skin was carried out from 7 POD on in both groups, and the pathomorphological changes were examined in both groups on 7 POD. RESULTS: The MC148 gene expression in rat skin of T group could be identified in 6 POH, and it reached the peak on 3 POD (A(MC148 mRNA) / A(beta_actin) = 0.86), and then subsided thereafter, but it maintained for 10 days. The survival time of the grafts in T group was (15.0 +/- 2.0) days, and it was significantly longer than that in C group (8.5 +/- 3.4) days, (P < 0.01). Gross and microscopic examination showed that the tail skin of T group appeared ruddy on 7 POD, with little leukocytic infiltration in subcutaneous tissue; it began to turn black after 12 to 20 PODs. On the other hand, the tail skin of C group began to turn black and to shed off on 7 POD, with evident leukocytic infiltration in subcutaneous tissue and dermis. CONCLUSION: Local transfection of MC148 gene may promote immunosuppression by inhibiting leukocytic infiltration after allogeneous skin transplantation.