CAJ | 학술논문

目的探讨局部转染基质细胞衍生因子-1α(SDF-1α)基因预防显微血管吻合术后血栓形成的效果. 方法 2014年6月至2015年4月,利用基因工程技术构建SDF-1α基因重组质粒.使用体质量300～350 g的SD大鼠60只,其中15只大鼠左下肢肌肉内分别注射重组SDF-1α质粒、空载质粒及生理盐水,注射前及注射后第3、7、14、28天,利用ELISA试剂盒检测血浆SDF-1α水平;另外45只大鼠随机分为3组,分别注射重组质粒、空载质粒及生理盐水.注射后第7天,将所有大鼠左侧股动脉挤压离断后行端端血管吻合.术后15 min、30 min及2h,利用通畅实验检测吻合口是否堵塞;术后2h取吻合口股动脉标本采用HE染色、扫描电镜检测血栓形成情况,RT-PCR检测相关基因转录水平. 结果大鼠下肢局部注射重组SDF-1α质粒后外周血SDF-1α水平短暂升高,第7天达到高峰,第28天基本恢复正常水平.术后2h,重组质粒组通畅率(60％)高于空载质粒组及生理盐水组(均为20％),差异具有统计学意义(P＜0.05).苏木素-伊红(HE)染色显示重组质粒组血栓堵塞面积百分比为(52.00±36.27)％,小于[生理盐水组(84.30±22.59)％和质粒对照组[(85.40±18.93％)],差异有统计学意义(P＜0.05).RT-PCR结果发现术后2h,重组质粒组内皮型一氧化氮合酶基因转录水平升高,差异有统计学意义(P＜0.05).扫描电镜提示重组质粒组血栓明显少于对照组.结论局部转染SDF-1α基因能够抑制显微血管吻合术后吻合口血栓形成,提高吻合口通畅率,可能与局部内皮型一氧化氮合酶转录水平增高有关.上述结果为显微外科血管吻合术后血栓的预防提供新的方法.
목적 탐토국부전염기질세포연생인자-1α(SDF-1α)기인예방현미혈관문합술후혈전형성적효과. 방법 2014년6월지2015년4월,이용기인공정기술구건SDF-1α기인중조질립.사용체질량300～350 g적SD대서60지,기중15지대서좌하지기육내분별주사중조SDF-1α질립、공재질립급생리염수,주사전급주사후제3、7、14、28천,이용ELISA시제합검측혈장SDF-1α수평;령외45지대서수궤분위3조,분별주사중조질립、공재질립급생리염수.주사후제7천,장소유대서좌측고동맥제압리단후행단단혈관문합.술후15 min、30 min급2h,이용통창실험검측문합구시부도새;술후2h취문합구고동맥표본채용HE염색、소묘전경검측혈전형성정황,RT-PCR검측상관기인전록수평. 결과 대서하지국부주사중조SDF-1α질립후외주혈SDF-1α수평단잠승고,제7천체도고봉,제28천기본회복정상수평.술후2h,중조질립조통창솔(60％)고우공재질립조급생리염수조(균위20％),차이구유통계학의의(P＜0.05).소목소-이홍(HE)염색현시중조질립조혈전도새면적백분비위(52.00±36.27)％,소우[생리염수조(84.30±22.59)％화질립대조조[(85.40±18.93％)],차이유통계학의의(P＜0.05).RT-PCR결과발현술후2h,중조질립조내피형일양화담합매기인전록수평승고,차이유통계학의의(P＜0.05).소묘전경제시중조질립조혈전명현소우대조조.결론 국부전염SDF-1α기인능구억제현미혈관문합술후문합구혈전형성,제고문합구통창솔,가능여국부내피형일양화담합매전록수평증고유관.상술결과위현미외과혈관문합술후혈전적예방제공신적방법.
Objective To evaluate the effect of local transfer of stromal cell-derived factor-1α (SDF-1α) gene on inhibiting microsurgical anastomotic thrombosis in a crushing model of rat femoral artery.Methods From June,2014 to April, 2015, sixty Sprague-Dawley rats were used in this study.Plasmid DNA encoding SDF-1α gene, empty plasmid, or saline was injected into the left femoral muscles in 15 rats.Plasma level of SDF-1α was tested by ELISA at 3, 7, 14 and 28 days after injection.The other 45 rats were divided randomly into 3 groups.Plasmid DNA encoding SDF-1α, empty plasmid or saline was injected in each group respectively.Seven days after injection, the left femoral artery was crushed, transected and repaired by end-to-end microsurgical anastomosis in each rat.Vascular patency was assessed by milk test at 15, 30 and 120 minutes after reperfusion.Thrombosis formation was assessed using H&E staining and scanning electron microscopy.RT-PCR was used to detect the expression of endothelial nitric oxide synthase, thrombomodulin, yon Willebrand factor and tissue type plas minogen activator.Results The plasma level of SDF-1α increased transiently, and peaked at 7 days after injection (P ＜ 0.05).The patency rates in SDF-1α group were significantly higher than that in control groups at 120 minutes (20％, 20％ and 60％ in three groups respectively) (P ＜ 0.05).Treatment of SDF-1α significantly reduced the area of thrombotic occlusion compared with controls (84.30 ± 22.59％, 85.40 ± 18.93％, 52.00 ± 36.27％ in saline, empty plasmid and SDF-1α groups, respectively) (P ＜ 0.05).RT-PCR results revealed that endothelial nitric oxide synthase was overexpressed in SDF-1α group than control groups (P ＜ 0.05).Conclusion The current study showed that local transfer of SDF-1α gene increases arterial patency and inhibits microsurgical anastomotic thrombosis in a crush model of rat femoral artery.This beneficial effect might be mediated through up-regulation of endothelial nitric oxide synthase.These findings provide a novel approach for inhibition of microsurgical anastomotic thrombosis.