CAJ | 학술논문

目的探讨联合应用骨髓间质细胞(bone marrow stromal cell,BMSC)与bcl-2基因治疗脑缺血的疗效,及其对碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)表达的影响.方法 40只Wistzx大鼠制作大脑中动脉闭塞模型,随机分为生理盐水对照组、bcl-2组、BMSC组和BMSC+bcl=2组,每组10只;每组再分为缺血再灌注后3 d和14 d亚组,每组5只.BMSC用溴脱氧尿苷(bromodeoxyuridine,BrdU)标记;实验大鼠行神经功能评分;脑内BMSC分布及数量、bFGF和Bcl-2蛋白表达采用免疫组化检测,凋亡细胞采用TUNEL染色检测.结果 BMSC+bcl-2组再灌注后3 d神经功能评分显著低于生理盐水对照组(P<0.05),14 d评分显著低于其余3组(P均<0.05).BMSC+bcl-2组和BMSC组梗死半球侧可见大量BrdU阳性BMSC细胞,BMSC+bcl-2组再灌注后3 d、14 d BrdU阳性细胞数均显著多于BMSC组(P均<0.05).BMSC+bcl-2组再灌注后3 d、14 d梗死侧Bcl-2蛋白表达水平显著高于其余3组(P均<0.05).BMSC+bcl-2组bFGF表达水平在各时间点均较其余3组显著增加(P均<0.05).BMSC+bcl-2组各时间点脑内凋亡细胞数量均较其余3组显著减少(P均<0.05).结论 BMSC与bcl-2基因均有治疗脑缺血作用,二者联合应用的效果显著优于二者单独应用,可显著改善大鼠神经功能,提高bFGF表达,其机制可能是bcl-2基因在脑内抗凋亡的同时,也抑制了BMSC凋亡.
목적 탐토연합응용골수간질세포(bone marrow stromal cell,BMSC)여bcl-2기인치료뇌결혈적료효,급기대감성성섬유세포생장인자(basic fibroblast growth factor,bFGF)표체적영향.방법 40지Wistzx대서제작대뇌중동맥폐새모형,수궤분위생리염수대조조、bcl-2조、BMSC조화BMSC+bcl=2조,매조10지;매조재분위결혈재관주후3 d화14 d아조,매조5지.BMSC용추탈양뇨감(bromodeoxyuridine,BrdU)표기;실험대서행신경공능평분;뇌내BMSC분포급수량、bFGF화Bcl-2단백표체채용면역조화검측,조망세포채용TUNEL염색검측.결과 BMSC+bcl-2조재관주후3 d신경공능평분현저저우생리염수대조조(P<0.05),14 d평분현저저우기여3조(P균<0.05).BMSC+bcl-2조화BMSC조경사반구측가견대량BrdU양성BMSC세포,BMSC+bcl-2조재관주후3 d、14 d BrdU양성세포수균현저다우BMSC조(P균<0.05).BMSC+bcl-2조재관주후3 d、14 d경사측Bcl-2단백표체수평현저고우기여3조(P균<0.05).BMSC+bcl-2조bFGF표체수평재각시간점균교기여3조현저증가(P균<0.05).BMSC+bcl-2조각시간점뇌내조망세포수량균교기여3조현저감소(P균<0.05).결론 BMSC여bcl-2기인균유치료뇌결혈작용,이자연합응용적효과현저우우이자단독응용,가현저개선대서신경공능,제고bFGF표체,기궤제가능시bcl-2기인재뇌내항조망적동시,야억제료BMSC조망.
Objective To investigate the efficacy of the combination of bone marrow stromal cells (BMSCs) and bcl-2 gene in the treatment cerebral ischemia taxi its effect on the expression of basic fibroblast growth factor (bFGF) in rats.Methods Forty Wistar rats were used to establish middle cerebral artery occlusion model. They were randomly divided into 4 groups: saline control, bcl-2, BMSCs and BMSCs + bcl-2 groups (n = 10 in each group). Every group was redivided into 3- and 14-day after reperfusion subgroups (n = 5 in each subgroup). Neurological scores of the experimental rats were assessed. BMSCs were labeled with bromodeoxyuridine (BrdU). The distribution and numbers of BMSCs, the expressions of Bcl-2 and bFGF were detected by immunohistochemistry, and apoptotic cells were detected with TUNEL staining in rat brain. Results The neurological score at day 3 after reperfusion in the BMSCs + bcl-2 group was significantly lower than that in the saline control group (P < 0. 05), and at day 14, it was significantly lower than that in the other 3 groups (all P <0. 05). A large number of BrdU-positive BMSCs were observed in the infarcted hemisphere in the BMSCs + bcl-2 and BMSCs groups. The numbers of BrdU-positive BMSCs at day 3 and 14 after reperfusion in the BMSCs + bcl-2 group were significantly higher than those in the BMSCs group (all P <0. 05). The expressions of Bcl-2 in the infarcted hemisphere at day 3 and 14 after reperfusion in BMSCs +hel-2 group wre significantly higher than those in the other 3 groups (all P <0. 05). The expressions of Bcl-2 at all time points were increased more significantly than those in the other 3 groups (all P <0. 05). The numbers of apoptosis in brain at all time points in the BMSCs + bcl-2 group were decreased more significantly than those in the other 3 groups (all P <0. 05). Conclusions Both BMSCs and bcl-2 genes have the therapeutic effect on cerebral ischemia. The efficacy of combination of both ,of them is significantly superior to monotherapy. They may significantly improve the neurological function and increase the expression of bFGF in rats. Its mechanism may be that bcl-2 genes have inhibited BMSCs apoptosis at the same time of anti-apoptosis in brain.