CAJ | 학술논문

目的观察自体骨髓单核细胞移植对大脑中动脉栓塞(MCAO)大鼠脑梗死体积、血脑屏障破坏程度及行为学评分等的影响,评价骨髓单核细胞移植对脑梗死的治疗价值. 方法将72只成年雄性SD大鼠按完全随机数字表法分为假手术组、模型组、生理盐水组及骨髓单核细胞移植组,后三组采用线栓法制作MCAO大鼠模型.采用梯度离心法分离大鼠股骨骨髓腔中的单核细胞,并通过流式细胞术检测CD45+细胞比例.将分离得到的自体骨髓单核细胞经颈静脉途径移植到骨髓单核细胞移植组大鼠体内,生理盐水组大鼠给予等体积的生理盐水注射.造模后72 h时伊文氏蓝染色及脑组织含水量检测显示血脑屏障的破坏情况,造模后1d、3d、7d时通过Zea-Longer评分评价大鼠神经功能变化,最后一次Zea-Longer评分结束后TTC染色检测大鼠脑梗死体积. 结果流式细胞术检测发现骨髓单核细胞中CD45+细胞的比例为91.2％.脑梗死体积、伊文氏蓝含量及脑组织含水量在模型组与生理盐水组中均较高,且相互比较差异无统计学意义(P＞0.05)；骨髓单核细胞移植组脑梗死体积、伊文氏蓝含量及脑组织含水量均明显下降,较模型组与生理盐水组差异均有统计学意义(P＜0.05).Zea-Longer评分显示,除假手术组外,其余大鼠均有不同程度的神经功能缺损；造模后7d时骨髓单核细胞移植组神经功能评分明显改善,与模型组及生理盐水组比较差异均有统计学意义(P＜0.05). 结论骨髓单核细胞分离过程简单,利用其移植治疗脑梗死可以显著减轻脑损伤程度并促进神经功能恢复,对脑梗死具有很高的治疗价值.
목적 관찰자체골수단핵세포이식대대뇌중동맥전새(MCAO)대서뇌경사체적、혈뇌병장파배정도급행위학평분등적영향,평개골수단핵세포이식대뇌경사적치료개치. 방법 장72지성년웅성SD대서안완전수궤수자표법분위가수술조、모형조、생리염수조급골수단핵세포이식조,후삼조채용선전법제작MCAO대서모형.채용제도리심법분리대서고골골수강중적단핵세포,병통과류식세포술검측CD45+세포비례.장분리득도적자체골수단핵세포경경정맥도경이식도골수단핵세포이식조대서체내,생리염수조대서급여등체적적생리염수주사.조모후72 h시이문씨람염색급뇌조직함수량검측현시혈뇌병장적파배정황,조모후1d、3d、7d시통과Zea-Longer평분평개대서신경공능변화,최후일차Zea-Longer평분결속후TTC염색검측대서뇌경사체적. 결과 류식세포술검측발현골수단핵세포중CD45+세포적비례위91.2％.뇌경사체적、이문씨람함량급뇌조직함수량재모형조여생리염수조중균교고,차상호비교차이무통계학의의(P＞0.05)；골수단핵세포이식조뇌경사체적、이문씨람함량급뇌조직함수량균명현하강,교모형조여생리염수조차이균유통계학의의(P＜0.05).Zea-Longer평분현시,제가수술조외,기여대서균유불동정도적신경공능결손；조모후7d시골수단핵세포이식조신경공능평분명현개선,여모형조급생리염수조비교차이균유통계학의의(P＜0.05). 결론 골수단핵세포분리과정간단,이용기이식치료뇌경사가이현저감경뇌손상정도병촉진신경공능회복,대뇌경사구유흔고적치료개치.
Objective To investigate the therapeutic role of autologous bone marrow mononuclear cells (BM-MNCs) in cerebral infarction by detecting the infarct volume, integrity of blood-brain barrier (BBB) and functional recovery in rats with middle cerebral artery occlusion (MCAO).Methods Seventy-two adult male SD rats were randomly divided into sham-operated group, vehicle group,normal saline treatment group and BM-MNC treatment group.MCAO in rats of the later 3 groups was induced by occlusion of the right middle cerebral artery using an intra luminal filament technique.BM-MNCs,selected from their femur,were isolated with gradient centrifugation,and the proportion of CD45 + cells was detected by flow cytometry before infusion.BM-MNCs were re-injected by jugular vein infusion.Infarct volumes were determined with 2,3,5-triphenyltetrazolium chloride (TTC) staining; the content of Evans blue and brain water content were detected to evaluate the damage of BBB; in addition,neurological deficits were also assessed by Zea-Longer tests. Results Flow cytometry revealed that the proportion of CD45 +cells in BM-MNCs was 91.2％.The infarct volumes,contents of Evans blue and brain water in rats of BM-MNC treatment group were significantly decreased as compared with those in rats of the normal saline treatment group and vehicle group (P＜0.05); however,no significant difference of those was noted between rats of the normal saline treatment group and vehicle group (P＞0.05).Except for the sham-operated group, all the other rats had neurological deficits of varied degrees; the neurological deficits in rats of the BM-MNC treatment group were obviously improved as compared with those in rats of the vehicle group and normal saline treatment group (P＜0.05). Conclusion The separation of BM-MNCs is simple; BM-MNCs can significantly reduce the extent of brain injury and promote recovery of neurological function in rats with cerebral infarction,which indicate that BM-MNCs may have high therapeutic value in cerebral infarction.