中国医药导报
中國醫藥導報
중국의약도보
CHINA MEDICAL HERALD
2015年
11期
11-14
,共4页
邬志坚%江慧玲%陈泳晖%刘宁平
鄔誌堅%江慧玲%陳泳暉%劉寧平
오지견%강혜령%진영휘%류저평
血管内皮生长因子%骨髓间充质干细胞%血管新生
血管內皮生長因子%骨髓間充質榦細胞%血管新生
혈관내피생장인자%골수간충질간세포%혈관신생
VEGF%BMSCs%Angiogenesis
目的：观察pEGFP-VEGF165质粒转染骨髓间充质干细胞(BMSCs)可否促进脑缺血大鼠血管新生。方法全骨髓贴壁法分离获取第三代BMSCs，当BMSCs生长汇合率为80%~90%时以脂转法转染pEGFP-VEGF165质粒。制作大鼠大脑中动脉闭塞(MCAO)模型,分为模型组、BMSCs移植组(BMSCs组)、转血管内皮生长因子(VEGF)基因BMSCs移植组(BMSCs/VEGF组)。于造模后7、14、21 d进行神经行为学评分，免疫组织化学染色检测脑缺血边缘区血管内皮细胞标志物CD34和VEGF的表达，激光共聚焦显微镜观察血管生成情况。结果BMSCs组与BMSCs/VEGF组大鼠神经功能(BRS)评分较模型组明显降低(P<0.01)，BMSCs/VEGF组BRS评分低于BMSCs组，但差异无统计学意义(P>0.05)。与非缺血侧比较，BMSCs组和BMSCs/VEGF组缺血侧微血管直径、微血管面积均较其显著增加(P<0.05或P<0.01)；BMSCs组和BMSCs/VEGF组微血管直径、微血管面积均较模型组显著增加(P<0.05)，但BMSCs组与BMSCs/VEGF组比较，差异无统计学意义(P>0.05)。7 d时BMSCs/VEGF组VEGF阳性细胞较BMSCs组VEGF表达显著增加(P<0.05)，其他时间点两组比较差异无统计学意义(P>0.05)；BMSCs/VEGF组CD34阳性表达在各时间点较BMSCs组有所增加，但差异无统计学意义(P>0.05)。结论经VEGF基因修饰的BMSCs移植可提高缺血区血管生成，仅在移植后7 d比单纯BMSCs移植VEGF表达升高有显著性差异，在神经行为学评分、微血管新生、CD34表达等方面与单纯BMSCs比较，无明显优势。原因可能与VEGF165质粒转染效率不高及脑血管新生机制复杂有关。
目的：觀察pEGFP-VEGF165質粒轉染骨髓間充質榦細胞(BMSCs)可否促進腦缺血大鼠血管新生。方法全骨髓貼壁法分離穫取第三代BMSCs，噹BMSCs生長彙閤率為80%~90%時以脂轉法轉染pEGFP-VEGF165質粒。製作大鼠大腦中動脈閉塞(MCAO)模型,分為模型組、BMSCs移植組(BMSCs組)、轉血管內皮生長因子(VEGF)基因BMSCs移植組(BMSCs/VEGF組)。于造模後7、14、21 d進行神經行為學評分，免疫組織化學染色檢測腦缺血邊緣區血管內皮細胞標誌物CD34和VEGF的錶達，激光共聚焦顯微鏡觀察血管生成情況。結果BMSCs組與BMSCs/VEGF組大鼠神經功能(BRS)評分較模型組明顯降低(P<0.01)，BMSCs/VEGF組BRS評分低于BMSCs組，但差異無統計學意義(P>0.05)。與非缺血側比較，BMSCs組和BMSCs/VEGF組缺血側微血管直徑、微血管麵積均較其顯著增加(P<0.05或P<0.01)；BMSCs組和BMSCs/VEGF組微血管直徑、微血管麵積均較模型組顯著增加(P<0.05)，但BMSCs組與BMSCs/VEGF組比較，差異無統計學意義(P>0.05)。7 d時BMSCs/VEGF組VEGF暘性細胞較BMSCs組VEGF錶達顯著增加(P<0.05)，其他時間點兩組比較差異無統計學意義(P>0.05)；BMSCs/VEGF組CD34暘性錶達在各時間點較BMSCs組有所增加，但差異無統計學意義(P>0.05)。結論經VEGF基因脩飾的BMSCs移植可提高缺血區血管生成，僅在移植後7 d比單純BMSCs移植VEGF錶達升高有顯著性差異，在神經行為學評分、微血管新生、CD34錶達等方麵與單純BMSCs比較，無明顯優勢。原因可能與VEGF165質粒轉染效率不高及腦血管新生機製複雜有關。
목적：관찰pEGFP-VEGF165질립전염골수간충질간세포(BMSCs)가부촉진뇌결혈대서혈관신생。방법전골수첩벽법분리획취제삼대BMSCs，당BMSCs생장회합솔위80%~90%시이지전법전염pEGFP-VEGF165질립。제작대서대뇌중동맥폐새(MCAO)모형,분위모형조、BMSCs이식조(BMSCs조)、전혈관내피생장인자(VEGF)기인BMSCs이식조(BMSCs/VEGF조)。우조모후7、14、21 d진행신경행위학평분，면역조직화학염색검측뇌결혈변연구혈관내피세포표지물CD34화VEGF적표체，격광공취초현미경관찰혈관생성정황。결과BMSCs조여BMSCs/VEGF조대서신경공능(BRS)평분교모형조명현강저(P<0.01)，BMSCs/VEGF조BRS평분저우BMSCs조，단차이무통계학의의(P>0.05)。여비결혈측비교，BMSCs조화BMSCs/VEGF조결혈측미혈관직경、미혈관면적균교기현저증가(P<0.05혹P<0.01)；BMSCs조화BMSCs/VEGF조미혈관직경、미혈관면적균교모형조현저증가(P<0.05)，단BMSCs조여BMSCs/VEGF조비교，차이무통계학의의(P>0.05)。7 d시BMSCs/VEGF조VEGF양성세포교BMSCs조VEGF표체현저증가(P<0.05)，기타시간점량조비교차이무통계학의의(P>0.05)；BMSCs/VEGF조CD34양성표체재각시간점교BMSCs조유소증가，단차이무통계학의의(P>0.05)。결론경VEGF기인수식적BMSCs이식가제고결혈구혈관생성，부재이식후7 d비단순BMSCs이식VEGF표체승고유현저성차이，재신경행위학평분、미혈관신생、CD34표체등방면여단순BMSCs비교，무명현우세。원인가능여VEGF165질립전염효솔불고급뇌혈관신생궤제복잡유관。
Objective To observe pEGFP-VEGF165 plasmid transfection BMSCs whether promote angiogenesis of focal ischemia rats. Methods The third generation of BMSCs were isolated by whole bone marrow adherence method. As BMSCs growth convergence rate reached 80%-90%, pEGFP-VEGF165 plasmid transferred by lipid transfer method. Rats were used to make MCAO model, including model group, BMSCs group, BMSCs/VEGF group. Neurological behavior score finished after model 7, 14, 21 d. Expression of vascular endothelial cell markers CD34 and VEGF were tested by immunohistochemistry staining. Angiogenesis was observed by confocal laser scanning microscopy. Results BRS scores in BMSCs group and BMSCs/VEGF group were decreased significantly than those that in model group (P<0.01), BRS scores in BMSCs/VEGF group were lower than those in BMSCs group, but the difference was not statistically significant (P>0.05). Compared with non-ischemic side, microvessel diameter and area increased significantly in ischemic side in BMSCs group and BMSCs/VEGF group (P< 0.05 or P < 0.01). Microvessel diameter and area in BMSCs group and BMSCs/VEGF group were higher than those in model group (P<0.05), but there was no statistically significant differ-ence between BMSCs group and BMSCs/VEGF group. At 7 d, the expression of VEGF and in BMSCs/VEGF group was significantly higher than that in BMSCs group (P< 0.05). There was no statistically significant difference at other time;CD34 expression in BMSCs/VEGF group was higher than that in BMSCs group, but there was no statistically significant difference (P>0.05). Conclusion BMSCs transplantation modified by VEGF gene can improve angiogenesis in ischemic area, VEGF expression in which have significant difference with pure BMSCs transplantation at post-transplant 7 d. Compared with pure BMSCs at neurological behavior score; angiogenesis; CD34 expression, there are no obvious ad-vantage. Reason may be related to VEGF165 plasmid transfection rate low and complex mechanism of brain angiogenesis.
