中华整形外科杂志
中華整形外科雜誌
중화정형외과잡지
CHINESE JOURNAL OF PLASTIC SURGERY
2009年
2期
120-124
,共5页
吴银生%沈江涌%姚明%李津宁%赵巍%苏荣%陆安民
吳銀生%瀋江湧%姚明%李津寧%趙巍%囌榮%陸安民
오은생%침강용%요명%리진저%조외%소영%륙안민
血管内皮生长因子%基因疗法%外科皮瓣
血管內皮生長因子%基因療法%外科皮瓣
혈관내피생장인자%기인요법%외과피판
Vascular endothelial growth factor%Gene therapy%Surgical flaps
目的 通过血管内皮生长因子基因对大鼠随意型皮瓣的转染,探讨基因治疗对不同时间断蒂的大鼠随意型皮瓣成活的影响.方法 以SD大鼠为实验模型制作背部随意型皮瓣,实验组注入脂质体包裹的PcDNAVEGF165(目的 基因组),对照组分别注入PcDNA(空白质粒组)和生理盐水(生理盐水组),于用药后1、3、5、7 d,每组每时相点分别随机选取10只断蒂,断蒂后7 d处死大鼠,观察下述指标:①皮瓣成活率.②皮瓣组织标本行常规HE染色检测平均微血管数目及内径.③行VEGF免疫组织化学染色检测VEGF表达情况.④取皮瓣组织标本在电镜下观察超微结构.结果 ①皮瓣成活率:1、3、5、7 d断蒂实验组皮瓣成活率分别为(45.45±12.24)%、(82.95±3.81)%、(85.00±3.38)%、(85.96±3.25)%.1 d断蒂实验组与对照组比较差异无统计学意义(P>0.05),3、5、7 d断蒂各实验组明显高于相应对照组(P<0.05),3、5、7 d断蒂各实验组则随着断蒂时间的延迟差异无统计学意义(P>0.05).②平均微血管数目及内径:各实验组与相应对照组比较差异有统计学意义(P<0.05).③各实验组VEGF染色深度明显高于对照组(P<0.05).④超微结构:实验组内有新生血管形成,内皮细胞内可见较多粗面内质网,线粒体等结构,组织内成纤维细胞增多,细胞合成代谢旺盛.结论 皮下注射脂质体介导VEGF基因可提高皮瓣成活率,促进早期断蒂,是一种简单,高效,经济,相对安全的基因治疗方法.
目的 通過血管內皮生長因子基因對大鼠隨意型皮瓣的轉染,探討基因治療對不同時間斷蒂的大鼠隨意型皮瓣成活的影響.方法 以SD大鼠為實驗模型製作揹部隨意型皮瓣,實驗組註入脂質體包裹的PcDNAVEGF165(目的 基因組),對照組分彆註入PcDNA(空白質粒組)和生理鹽水(生理鹽水組),于用藥後1、3、5、7 d,每組每時相點分彆隨機選取10隻斷蒂,斷蒂後7 d處死大鼠,觀察下述指標:①皮瓣成活率.②皮瓣組織標本行常規HE染色檢測平均微血管數目及內徑.③行VEGF免疫組織化學染色檢測VEGF錶達情況.④取皮瓣組織標本在電鏡下觀察超微結構.結果 ①皮瓣成活率:1、3、5、7 d斷蒂實驗組皮瓣成活率分彆為(45.45±12.24)%、(82.95±3.81)%、(85.00±3.38)%、(85.96±3.25)%.1 d斷蒂實驗組與對照組比較差異無統計學意義(P>0.05),3、5、7 d斷蒂各實驗組明顯高于相應對照組(P<0.05),3、5、7 d斷蒂各實驗組則隨著斷蒂時間的延遲差異無統計學意義(P>0.05).②平均微血管數目及內徑:各實驗組與相應對照組比較差異有統計學意義(P<0.05).③各實驗組VEGF染色深度明顯高于對照組(P<0.05).④超微結構:實驗組內有新生血管形成,內皮細胞內可見較多粗麵內質網,線粒體等結構,組織內成纖維細胞增多,細胞閤成代謝旺盛.結論 皮下註射脂質體介導VEGF基因可提高皮瓣成活率,促進早期斷蒂,是一種簡單,高效,經濟,相對安全的基因治療方法.
목적 통과혈관내피생장인자기인대대서수의형피판적전염,탐토기인치료대불동시간단체적대서수의형피판성활적영향.방법 이SD대서위실험모형제작배부수의형피판,실험조주입지질체포과적PcDNAVEGF165(목적 기인조),대조조분별주입PcDNA(공백질립조)화생리염수(생리염수조),우용약후1、3、5、7 d,매조매시상점분별수궤선취10지단체,단체후7 d처사대서,관찰하술지표:①피판성활솔.②피판조직표본행상규HE염색검측평균미혈관수목급내경.③행VEGF면역조직화학염색검측VEGF표체정황.④취피판조직표본재전경하관찰초미결구.결과 ①피판성활솔:1、3、5、7 d단체실험조피판성활솔분별위(45.45±12.24)%、(82.95±3.81)%、(85.00±3.38)%、(85.96±3.25)%.1 d단체실험조여대조조비교차이무통계학의의(P>0.05),3、5、7 d단체각실험조명현고우상응대조조(P<0.05),3、5、7 d단체각실험조칙수착단체시간적연지차이무통계학의의(P>0.05).②평균미혈관수목급내경:각실험조여상응대조조비교차이유통계학의의(P<0.05).③각실험조VEGF염색심도명현고우대조조(P<0.05).④초미결구:실험조내유신생혈관형성,내피세포내가견교다조면내질망,선립체등결구,조직내성섬유세포증다,세포합성대사왕성.결론 피하주사지질체개도VEGF기인가제고피판성활솔,촉진조기단체,시일충간단,고효,경제,상대안전적기인치료방법.
Objective To investigate the effect of VEGF gene on the random flap survival after pedicle division at different time in rats. Methods The random-pattern flaps were formed on the back of the 120 SD rats. PcDNAVEGF165(gene) wrapped with liposome was'injected into the flaps in experimental group(n= 40). The flaps in the two control groups were injected with PcDNA (n = 40) or saline( n = 40). 1,3,5,7days after injection, 10 rats in each group were randomly selected to performed pedicle division. 7 days after pedicle division, the rats were killed to measure the flap survival rate. The microvessels was studied by histologic examination. The expression of VEGF was assessed by imunohistpchemical staining. The flaps were also examined under the electron uhrastrueture microscopy. Results ①Flap survival rote after pedicle division in experimental group at 1 day,3 days,5 days,7 days after injection,were (45.45 ±12.24)% ,(82.95± 3.81)% ,(85.00± 3.38) %, (85.96±3.25) %, respectively. The flap survival rates were significant different between experimental group and the control groups at 3, 5, 7 days after injection (P<0.05), but not at I days after injection (P> 0.05).② The average microvascular diameter and number in experimental group were significantly higher than those in control groups(P<0.05). ③The expression of VEGF in experimental group was significantly higher than that in the two control groups(P<0.05).④Ultrastrcture study showed more angiogenesis in experimental group. Conclusions Subcutaneous injection of liposome-mediated VEGF gene can increase the survival rate of flap with early pedicle division. It is a simple, efficient, economic, and the relatively safe gene therapy.
