异体复合组织移植皮肤淋巴引流隔绝动物模型的建立

异体复合组织移植皮肤淋巴引流隔绝动物模型的建立[摘要]目的：通过sd大鼠下腹部游离皮瓣及皮管的移植，建立大鼠异体复合组织移植皮肤淋巴引流隔绝模型。方法：7～8周龄雄性sd大鼠切取下腹部皮瓣并游离行同种异体移植，皮肤隔离组（a 组）：将供体皮瓣边缘对合、缝合成管状，底边留血管蒂出口用隔离器隔离皮肤，显微外科吻合血管，皮管缝合于隔离器，隔离器固定于受体；皮肤未隔离组（b组）：将供体游离皮瓣卷制成管状，显微外科吻合血管并将供受体皮肤对位缝合；空白对照组（c组）：行下腹部游离皮瓣异体移植。大体观察每组皮瓣或皮管存活时间，于术后3、5、7、14、28、35d取移植物皮肤组织行he染色,观察病理学变化。结果：a组：皮管存活时间为32（34，32）天；b组：皮管存活时间为15（16,15）天；c组：皮瓣存活时间为6（7,6）天。a组与c组、b组与c组、a组与b组存活率差异均有统计学意义（p＜0.05）。结论：本实验成功建立了大鼠的异体复合组织移植淋巴引流隔绝模型,并证实了隔绝皮肤淋巴引流能有效延长移植物的存活时间。

[关键词]异体复合组织移植；免疫抑制；动物模型；淋巴引流the establishing of composite tissue allograft skin lymphatic vessels isolation animal model in rats

liu sha,pei jiao-miao,li yang, cao jiao,yang yang，li bing，xia wei, guo shu-zhong

(institute of plastic surgery, xijing hospital, fourth

military medical university, xi’an 710032, shaanxi,china) abstract: objective to establish lymphatic vessels isolation animal model through vascularized hypogastric flap between sd rats. methods 18 couples of sd rats(7-8weeks) were randomly divided into 3 groups. group a(skin lymphatic vessles isolation): the free tube made of vascularized free flap located on the surface of the recipient through the isolation device after vascular anastomosis. group b(tube) : the free tube made of vascularized free flap sutured on the recipient lower abdomen directly after vascular anastomosis. group c(control) free flap allotransplantation. observe the survive ratio of animal and rejection time of composite tissue flap, histologic outcomes on the day 3,5,7,14,28, and 35 after transplantation. results the survival times of grafts in group a are 32（34, 32）days and group b were 15（16,15）days. by contrast,the survival time of group c was 6（7,6）days. the difference among these three groups were significant. conclusions this experiment successfully established lymphatic vessels isolation animal model in sd rats. the prolonged survival times of the grafts in this animal model had been proved.

key words: composite tissue allotransplantation；

immunosuppression；animal model; lymph draining

器官移植已成功应用于临床50多年，然而异体复合组织移植却仍处于研究阶段，难以在临床上推广应用。皮肤的强抗原性和缺少有效安全的免疫抑制方案成为限制异体复合组织移植发展的主要

因素[1-2]。探索皮肤强抗原性的机制，并通过寻找靶点进行干预，来降低机体对异体皮肤移植物的免疫排斥成为研究热点[2]。血液循环系统及淋巴引流系统是机体排斥同种异体移植物的重要组织

结构，然而哪个更为重要仍存在争议[3]。在异体皮肤及角膜的移植研究中，证实引流淋巴结及淋巴引流至关重要，对切除引流淋巴结的豚鼠进行相应引流部位的异体皮肤移植后，异体皮肤存活时间延长[4]，角膜的langerhans等骨髓来源的细胞[5-6]在移植后可以迁移至局部引流淋巴结，提呈角膜的异体抗原，引发免疫排斥反应[7]，因此阻断异体复合组织移植物的皮肤淋巴引流与受体相接触，有效的切断移植物中的抗原提呈细胞通过淋巴管引流至引流淋巴结，从而阻止其激活t细胞，无法启动免疫排斥反应，进而可能诱导免疫耐受，延长移植物的存活时间。本实验拟建立大鼠下腹部异体复合组织移植皮肤淋巴引流隔绝模型，该模型的异体移植物仅有血液循环系统而无淋巴引流系统，通过观察移植物的存活情况来评估淋巴引流系统对启动移植排斥的重要性。

1 材料和方法

1.1 实验动物与分组：随机选取清洁级体重280～300g sd雄性大鼠36只（供受体各18只），由第四军医大学实验动物中心提供；