【摘要】：目的：檢測分析攜人血管內(nèi)皮生長因子-165基因骨髓間充質(zhì)干細(xì)胞對(duì)大鼠缺血后肢的治療效果,為下肢缺血的臨床治療提供新思路。 方法：本實(shí)驗(yàn)設(shè)立三個(gè)實(shí)驗(yàn)組,分別為轉(zhuǎn)染hVEGF-165基因BMSC組、BMSC組、空白對(duì)照組,每組有效樣本8例,共48條肢體。采用結(jié)扎SD大鼠腹主動(dòng)脈分叉處上方建立缺血模型,并通過術(shù)后觀察、DSA血管造影、測股動(dòng)脈血流速度及阻力檢測鑒定；通過慢病毒介導(dǎo)hVEGF-165基因轉(zhuǎn)染大鼠骨髓間充質(zhì)干細(xì)胞,GFP熒光蛋白標(biāo)記,熒光顯微鏡下檢測hVEGF-165基因轉(zhuǎn)染率；再采用大鼠后肢肌肉多點(diǎn)注射方法將1X106/ml濃度的干細(xì)胞移植到缺血的肢體內(nèi),一個(gè)月后通過觀察、DSA血管造影、測股動(dòng)脈血流速度及血流阻力檢測療效。 結(jié)果：1、熒光顯微鏡下檢測慢病毒介導(dǎo)hVEGF-165基因轉(zhuǎn)染BMSCs陽性率可達(dá)85%以上；2、采用結(jié)扎腹主動(dòng)脈法制作缺血模型,術(shù)后30天所有大鼠出現(xiàn)跛行,足趾壞死,絕大部分大鼠肢體出現(xiàn)缺血性潰瘍。彩色多普勒超聲、后肢血管造影等檢查顯示結(jié)扎術(shù)后實(shí)驗(yàn)大鼠后肢血流、阻力指數(shù)顯著下降,血管密度顯著減少。3、移植術(shù)后30天檢測療效,大鼠后肢血流速度、阻力指數(shù)與微血管密度轉(zhuǎn)基因BMSC組、BMSC組均明顯高于空白對(duì)照組,而轉(zhuǎn)基因BMSC組高于BMSC組(p0.01)。 結(jié)論：慢病毒載體介導(dǎo)hVEGF-165基因轉(zhuǎn)染BMSC陽性率高；采用SD大鼠腹主動(dòng)脈分叉處上方結(jié)扎法制作后肢缺血模型缺血效果明顯,持續(xù)時(shí)間長；相同移植條件下的攜hVEGF-165基因BMSCs、單純BMSCs均能改善缺血后肢的血供,但轉(zhuǎn)染hVEGF165基因BMSCs促血管新生能力更有優(yōu)勢。
[Abstract]:Aim: to investigate the therapeutic effect of bone marrow mesenchymal stem cells carrying human vascular endothelial growth factor-165 gene on hindlimb ischemia in rats. Methods: three experimental groups were set up in this experiment. They were hVEGF-165 gene transfected BMSC group and blank control group. There were 8 effective samples in each group with 48 limbs in each group. The ischemia model was established by ligating the abdominal aortic bifurcation of SD rats. The blood flow velocity and resistance of femoral artery were measured and identified by observing DSA angiography after operation. The hVEGF-165 gene was transfected into rat bone marrow mesenchymal stem cells (BMSCs) by lentivirus-mediated transfection with GFP fluorescent protein. The transfection rate of hVEGF-165 gene was detected by fluorescence microscope. The stem cells with 1X106/ml concentration were transplanted into ischemic limbs by multi-point injection of rat hindlimb muscle. One month later, the blood flow velocity and resistance of femoral artery were measured by observing DSA angiography. Results under the fluorescence microscope, the positive rate of lentivirus mediated hVEGF-165 gene transfection BMSCs was over 85%. The ischemic model was made by ligation of abdominal aorta. 30 days after operation, all rats showed claudication and toe necrosis. Most rats had ischemic ulcers in their limbs. Color Doppler ultrasound, hindlimb angiography and other examinations showed that the blood flow of the hind limb of the experimental rats after ligation was significantly decreased, the resistance index was significantly decreased, the blood vessel density was significantly decreased by 0.3, the curative effect was detected 30 days after transplantation, and the blood flow velocity of the hind limb of the rats was measured. The resistance index and microvessel density in BMSC group were significantly higher than those in control group, while those in transgenic BMSC group were higher than those in BMSC group (p0.01). Conclusion: the positive rate of hVEGF-165 gene transfection with lentivirus vector is high, and the ischemic effect of hind limb ischemia model made by ligation above the abdominal aorta bifurcation of SD rats is obvious and lasting for a long time. BMSCs, carrying hVEGF-165 gene could improve the blood supply of hind limb after ischemia, but transfection of hVEGF165 gene BMSCs had more advantages in promoting angiogenesis.
【學(xué)位授予單位】：遵義醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R329

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