中华神经医学杂志
中華神經醫學雜誌
중화신경의학잡지
CHINESE JOURNAL OF NEUROMEDICINE
2014年
3期
252-256
,共5页
任瑞芳%黄良国%黄名璐%蒋国红%白洁
任瑞芳%黃良國%黃名璐%蔣國紅%白潔
임서방%황량국%황명로%장국홍%백길
间质干细胞%慢病毒属%脑源性神经营养因子%细胞迁移分析
間質榦細胞%慢病毒屬%腦源性神經營養因子%細胞遷移分析
간질간세포%만병독속%뇌원성신경영양인자%세포천이분석
Mesenchymal stem cell%Lentivirus%Brain-derived neurotrophic factor%Cell migration
目的 观察脑源性神经营养因子(BDNE)基因重组慢病毒转染骨髓间质干细胞(BMSCs)后,被移植入大鼠侧脑室内,向对侧脑出血灶周围组织的迁移情况. 方法 将60只脑出血模型大鼠按随机数字表法分为磷酸盐缓冲液(PBS)组、骨髓间质干细胞(BMSCs)组、骨髓间质干细胞-绿色荧光蛋白组(BMSCs-EGFP)、脑源性神经营养因子(BMSCs-EGFP-BDNF)组,每组15只.分别向侧脑室注射PBS,BMSCs,慢病毒(LV)转染的EGFP,LV转染的BDNF-EGFP,在术后7d、14d、21d采用Westen blotting检测各组BDNF在BMSCs中的蛋白表达,免疫荧光检测Brdu标记的BMSCs,EGFP及BDNF. 结果 Western blotting检测BMSCs-EGFP-BDNF组BDNF蛋白表达明显多于BMSCs组及BMSCs-EGFP组(P＜0.05);细胞爬片BMSCs-EGFP-BDNF组BDNF荧光表达明显高于BMSCs组及BMSCs-EGFP组;脑组织切片免疫荧光单标显示BMSCs-EGFP-BDNF组迁移至脑出血灶周围组织的BMSCs阳性细胞数明显多于BMSCs组及BMSCs-EGFP组(P＜0.05),BMSCs组与BMSCs-EGFP组除7d外,其余比较差异无统计学意义(P＞0.05). 结论 基因重组慢病毒修饰的BMSCs中BDNF表达增高,提示BDNF对BMSCs向脑出血灶周围组织迁移具有保护作用.
目的 觀察腦源性神經營養因子(BDNE)基因重組慢病毒轉染骨髓間質榦細胞(BMSCs)後,被移植入大鼠側腦室內,嚮對側腦齣血竈週圍組織的遷移情況. 方法 將60隻腦齣血模型大鼠按隨機數字錶法分為燐痠鹽緩遲液(PBS)組、骨髓間質榦細胞(BMSCs)組、骨髓間質榦細胞-綠色熒光蛋白組(BMSCs-EGFP)、腦源性神經營養因子(BMSCs-EGFP-BDNF)組,每組15隻.分彆嚮側腦室註射PBS,BMSCs,慢病毒(LV)轉染的EGFP,LV轉染的BDNF-EGFP,在術後7d、14d、21d採用Westen blotting檢測各組BDNF在BMSCs中的蛋白錶達,免疫熒光檢測Brdu標記的BMSCs,EGFP及BDNF. 結果 Western blotting檢測BMSCs-EGFP-BDNF組BDNF蛋白錶達明顯多于BMSCs組及BMSCs-EGFP組(P＜0.05);細胞爬片BMSCs-EGFP-BDNF組BDNF熒光錶達明顯高于BMSCs組及BMSCs-EGFP組;腦組織切片免疫熒光單標顯示BMSCs-EGFP-BDNF組遷移至腦齣血竈週圍組織的BMSCs暘性細胞數明顯多于BMSCs組及BMSCs-EGFP組(P＜0.05),BMSCs組與BMSCs-EGFP組除7d外,其餘比較差異無統計學意義(P＞0.05). 結論 基因重組慢病毒脩飾的BMSCs中BDNF錶達增高,提示BDNF對BMSCs嚮腦齣血竈週圍組織遷移具有保護作用.
목적 관찰뇌원성신경영양인자(BDNE)기인중조만병독전염골수간질간세포(BMSCs)후,피이식입대서측뇌실내,향대측뇌출혈조주위조직적천이정황. 방법 장60지뇌출혈모형대서안수궤수자표법분위린산염완충액(PBS)조、골수간질간세포(BMSCs)조、골수간질간세포-록색형광단백조(BMSCs-EGFP)、뇌원성신경영양인자(BMSCs-EGFP-BDNF)조,매조15지.분별향측뇌실주사PBS,BMSCs,만병독(LV)전염적EGFP,LV전염적BDNF-EGFP,재술후7d、14d、21d채용Westen blotting검측각조BDNF재BMSCs중적단백표체,면역형광검측Brdu표기적BMSCs,EGFP급BDNF. 결과 Western blotting검측BMSCs-EGFP-BDNF조BDNF단백표체명현다우BMSCs조급BMSCs-EGFP조(P＜0.05);세포파편BMSCs-EGFP-BDNF조BDNF형광표체명현고우BMSCs조급BMSCs-EGFP조;뇌조직절편면역형광단표현시BMSCs-EGFP-BDNF조천이지뇌출혈조주위조직적BMSCs양성세포수명현다우BMSCs조급BMSCs-EGFP조(P＜0.05),BMSCs조여BMSCs-EGFP조제7d외,기여비교차이무통계학의의(P＞0.05). 결론 기인중조만병독수식적BMSCs중BDNF표체증고,제시BDNF대BMSCs향뇌출혈조주위조직천이구유보호작용.
Objective To observe the migration ofmesenchymal stem cells (MSCs) transfected with recombinant lentiviral vectors carried brain derived neurotrophic factor (BDNF) gene from lateral ventricle to intracerebral hemorrhage stove in rats and to discuss the protective effect of BDNF on MSCs.Methods Intracerebral hemorrhagic models were constructed in 60 SD rats and randomly divided into 4 groups:phosphate buffer solution (PBS) group,BMSCs group,BMSCs-enhanced green fluorescent protein (EGFP) group and BMSCs-EGFP-BDNF group (n=15); PBS,BMSCs,lentiviral vector (LV)carried EGFP and LV carried BDNF-EGFP were,respectively,injected into the lateral cerebral ventricle of each group; 7,14 and 21 d after the injection,BDNF protein expression in the BMSCs of each group was detected by Western blotting and immunofluorescence; the migration of BMSCs from lateral ventricle to intracerebral hemorrhage stove was observed by signal labeling immunofluorescent staining.Results Western blotting and immunofluorescence showed that the BDNF protein expression in the MSCs-EGFP-BDNF group was significantly higher than that in the MSCs group and MSCs-EGFP group (P＜0.05); signal labeling immunofluorescent staining of brain tissue section indicated that the MSCs-EGFP-BDNF group had significantly larger number of BMSCs-positive cells which migrated to the surrounding issues of intracerebral hemorrhage stove than MSCs group and MSCs-EGFP group (P＜0.05),while there were no significant differences between the latter two except on the 7th d of establishment of models.Conclusion BMSCs modified with recombinant LV carried BDNF gene have high expression level of BDNF,indicating that BDNF performs a protective effect on BMSCs.
