VEGF-C基因修饰的淋巴结移植促进淋巴内皮细胞再生的初步研究  

A PRELIMINARY STUDY ON VASCULAR ENDOTHELIAL GROWTH FACTOR C GENE MODIFIED LYMPH NODE TRANSPLANTATION IN PROMOTING PROLIFERATION OF LYMPHATIC ENDOTHELIAL CELLS

作　　者：陆南杭[1] 张勇[1] 冯自豪[1] 亓发芝[1] 

机构地区：[1]复旦大学附属中山医院整形外科,上海200032

出　　处：《中国修复重建外科杂志》2013年第5期619-623,共5页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：上海市教育委员会科研创新项目(11ZZ06)~~

摘　　要：目的探讨采用VEGF-C基因修饰的淋巴结移植对移植淋巴结周围组织内淋巴内皮细胞再生的影响。方法取成年雄性SD大鼠36只,体重200.1～271.5 g,选取一侧采用带血管蒂淋巴结同侧原位移植方法制备腋窝淋巴结移植模型后,随机分为两组(n=18),分别将1.5×108PFU带Flag标签的VEGF-C基因过表达腺病毒载体及空载腺病毒载体注射入实验组及对照组大鼠移植淋巴结中。术后3 d,免疫荧光染色观察淋巴结内带Flag标签蛋白的表达情况;术后1、2、4周,免疫荧光染色观察移植淋巴结周围组织内Prox-1蛋白的表达并计算其荧光密度值;术后2、4周,通过测定患肢皮下注射偶氮蓝后大鼠血液在620 nm波长下吸光度(A)值,观察淋巴回流功能改善情况,以正常大鼠(正常组)及淋巴结移植大鼠(空白对照组)作为对照。结果术后3 d实验组及对照组移植淋巴结内均可检测到带Flag标签蛋白的表达。术后1、2、4周,实验组移植淋巴结周围组织内Prox-1蛋白荧光密度值呈进行性增加,且均明显强于对照组(P<0.05)。正常组及空白对照组大鼠血液A值分别为0.539±0.020及0.151±0.007。术后2周实验组及对照组A值分别为0.170±0.011及0.168±0.010,4周时分别为0.212±0.016及0.197±0.006,均显著低于正常组(P<0.05),但与空白组比较差异无统计学意义(P>0.05);实验组及对照组间比较,差异亦无统计学意义(P>0.05)。结论 VEGF-C基因修饰的淋巴结移植能显著促进移植淋巴结周围组织内淋巴内皮细胞的再生,但在4周内不能改善大鼠患肢淋巴回流功能。Objective To investigate the effects of vascular endothelial growth factor C （VEGF-C） gene modified lymph nodes on promoting proliferation of lymphatic endothelial cells in the surrounding tissues. Methods Thirty-six Sprague Dawley rats, weighing 200.1-271.5 g, were randomly divided into 2 groups （n=lS）. After the in situ axillary lymph nodes transplantation models were established in both groups, 1.5 x l0s PFU Ad-VEGF-C-Flag and Ad-Flag were injected into the transplanted lymph nodes in experimental group and control group, respectively. At 3 days after injection, the axillary lymph nodes were harvested to observe the expression of Flag; at 1, 2, and 4 weeks after injection, the axillary lymph nodes and the surrounding tissues were harvested to observe the expression of Prxo-1 protein and to calculate the fluorescence density; at 2 and 4 weeks after injection, the absorbance （A） value of treated blood at 620 nm was calculated to observe lymphatic back- flow function improvement; the rats without treatment served as normal control group, and the rats with in situ axillary lymph nodes transplantation model served as blank control group. Results At 3 days after injection, the expression of Flag could be detected in experimental group and control group. The fluorescence density of Prox-1 protein in experimental group increased at 1, 2, and 4 weeks, and it was significantly higher than that in control group （P 〈 0.05）. The A values of normal control group and blank control group were 0.539 ± 0.020 and 0.151± 0.007, respectively. The A values of experimental group and control group were 0.170 ± 0.011 and 0.168 ± 0.010 at 2 weeks, and 0.212± 0.016 and 0.197 ± 0.006 at 4 weeks, which were significantly lower than those of normal control group （P 〈 0.05）, but no significant difference was found when compared with blank control group, and between the experimental group and control group （P 〉 0.05）. Conclusion The VEGF-C gene modified lymph nodes can stimulate the proliferation of lympha

关 键 词：淋巴结移植 VEGF-C 淋巴内皮细胞 大鼠 

分 类 号：R654.7[医药卫生—外科学]