回族药扎里奴思方对骨髓间充质干细胞由血管迁移脑内的影响

Effect of Hui Medicine of Zhali Nusi Recipe on Bone Marrow Mesenchymal Stem Cells from Vascular to Brain Tissue After Transplantation

目的:观察扎里奴思方调控血脑屏障(BBB)通透性对骨髓间充质干细胞(BMSCs)经由血管迁移脑内的影响。方法:250只SD大鼠随机分为假手术组、模型组、扎方组、移植组和联合组,除假手术组10只外,其余各组15只,并采用线栓法制备大鼠大脑中动脉阻塞(MCAO)模型,体外全骨髓贴壁法培养及扩增BMSCs;大鼠ig给药(14.6 g·kg-1·d-1),BMSCs悬浮液经颈内动脉移植入脑(2×106/200μL);移植后1,3,7,14 d取材,干湿重法检测脑含水量,TTC法检测脑梗死面积,荧光分光光度计法检测伊文思蓝(EB)含量。结果:模型组大鼠脑含水量、脑梗死面积及EB含量均较假手术组显著增加(P<0.01);与模型组比较,扎方、移植及联合各3,7,14 d组脑含水量降低(P<0.01),各时间点脑梗死面积减小(P<0.01,P<0.05),扎方、联合各组及移植3,7 d组EB含量降低显著(P<0.01);与移植组比较,扎方1 d组脑含水量降低(P<0.05),1,7,14 d组EB含量降低(P<0.01,P<0.05),联合各组脑含水量、脑梗死面积及EB含量均降低,以1,14 d降低明显(P<0.01);扎方与联合组比较,联合各组上述指标均降低,以7,14 d组降低明显(P<0.01,P<0.05);同组间比较,均以7 d组变化显著,呈先增后减趋势,14 d有明显改善(P<0.01)。结论:脑缺血再灌注后7 d大鼠BBB损伤较1,3 d加重,14 d时有不同程度恢复;扎里奴思方可调控脑缺血再灌注损伤(CIRI)后BBB通透性,促进BMSCs经由血管进入脑内更好的发挥脑保护作用,两者联合作用显著,且促进作用随损伤时间延长呈增强趋势。

Objective：To observe the effect of Zhali Nusi recipe （ZLNS） on regulating blood-brain barrier （BBB） permeability and promoting the grafting of bone marrow mesenchymal stem cells （BMSCs） from vascular to brain tissue after the transplantation.Method：SD rats were randomly divided into sham-operated group,model group,ZLNS,BMSCs transplantation group and ZLNS combined with BMSCs group.Middle cerebral artery occlusion （MCAO） model was duplicated with nylon thread.BMSCs were cultured and amplified by the whole bone marrow adherence method.The rats were intragastrically administrated with ZLNS （14.6 g·kg^-1· d^-1）,BMSCs suspension solution were transplanted into brain through carotid artery （2 × 10v6/200 μL）.Brain of rats was taken out at 1,3,7,14 days after transplantation.Brain water content （BWC）,brain infarct area ratio （BIAR） and Evans Blue （EB） leakage were detected by dry-weight assay,TTC and fluorescence spectrophotometry,respectively.Result：The BWC,BIAR and EB in model group increased obviously than the sham-operated group （P 〈 O.01）.Compared with the model group,the BWC at 3,7,14 days decreased obviously,the EB leakage decreased at 3,7 days in ZLNS,transplantation and combination groups （P 〈 0.01）,the BIAR decreased in all groups （P 〈 0.01,P 〈0.05）.Compared with the transplantation group,the BWC at 1 day and EB at 1,7,14 days in ZLNS groups decreased （P 〈 0.01,P 〈 0.05）,all above indicators in combination groups decreased and the results were obvious at 1,14 days （P 〈0.01）.Compared with the ZLNS groups,all above indicators decreased in combination group and the results were obvious at 7,14 days （P 〈 0.01,P 〈 0.05）.All changes were remarkably serious at 7 days and obviously improved at 14 days being increased firstly and then decreased after cerebral ischemia in each groups （P 〈 0.01）.Conclusion：The injury at 7 days after cerebral ischemia reperfusion was serious than that at 1,3 days,and there was some improvement at 14 days in rats after cerebral ischemia reperfusion.The ZLNS could regulate the permeability of BBB,promote the grafting of BMSCs from vascular to brain tissue after the transplantation.It would play a better role in brain protection by the combination ZLNS and BMSCs transplantation.