摘要
目的研究Gja5基因表达在小鼠下肢新生动脉生成中的保护作用。方法将168只Gja5基因敲除小鼠分为CX40基因敲除实验组(Gja5^(-/-)组),CX40杂合子组(Gja5^(+/-)组)和对照组野生型小鼠(Gja5(+/+)组)3组,每组56只。另选择增强型绿色荧光蛋白(EGFP)标记的Gja5基因小鼠20只。在荧光显微镜下观察EGFP标记的Gja5基因在小鼠动脉的表达,建立小鼠下肢股动脉闭塞模型,应用激光多普勒血流仪(LDF)动态监测股动脉结扎术前,术后当天及术后1、3、7、14、21d下肢缺血区域的血流恢复情况,同时使用RT-PCR法检测Gja5基因在小鼠下肢腓肠肌的表达。结果 EGFP标记的Gja5基因(CX40基因)表达在小鼠的脑动脉和股动脉上。在股动脉结扎术后1、3、7、14、21d,Gja5^(-/-)组小鼠的息肢血供较Gja5^(+/+)组明显减少。在股动脉结扎术后3d时,Gja5基因在小鼠下肢腓肠肌的表达升高了约4倍,术后7d时逐渐回到基线水平。结论 Gja5基因表达在小鼠下肢新生动脉生成中发挥了重要的保护作用。
Objective To investigate the relationship between the gene expression of Gja5 （connexin-40, CX-40） and ar-teriogenesis on the hindlimb in mice. Methods One hundred and sixty eight mice were used in the study, including 56 CX40 knockout mice （Gja5-/- group）,56 CX40 heterozygous mice （Gja5＋/-group） and 56 wild-type mice （Gja5＋/＋ group）. The hindlimb blood flow blockage was induced by femoral arteries occlusion （FAO）. The scanning Laser Doppler Flow Imager was used to ob-serve the expression of Gja5 in Gja5 EGFP reporter animals through fluorescent microscope. Gja5 gene expression was also de-tected in gastrocnemicus（GC）muscle by real-time RT-PCR before and 1, 3, 7, 14, 21d after FAO. Results The Gja5 gene was expressed in mouse arteries including cerebral artery and femoral artery. There were significant reductions for hindlimb perfusion in Gja5-/- mice compared to Gja5＋/＋ mice at d1, 3, 7, 14 and 21 after FAO. The expression levels of Gja5 in Gja5＋/- mice and Gja5＋/＋mice in GC muscle were both increased 4-fold in the ischemic hindlimb at d3 after FAO. Then the expression levels of Gja5 re-turned to baseline values at d7 after FAO. Conclusion Arterial Gja5 expression may be involved in the arteriogenesis of ischemic hindlimb of mice.
出处
《浙江医学》
CAS
2013年第8期619-622,共4页
Zhejiang Medical Journal
基金
浙江省钱江人才计划D类项目(QJD1202017)
浙江省医药卫生平台重点资助计划项目(2013ZDA003)
