异氟烷干预下人羊膜间充质干细胞移植对大鼠脑梗死的影响

目的:观察人羊膜间充质干细胞(AM-MSCs)移植同时予以异氟烷治疗对脑梗死大鼠神经功能恢复的影响。方法:大鼠100只,采用线栓法建立大脑中动脉阻塞脑梗死模型,随机分成假手术组、脑梗死组、AMMSCs移植组、异氟烷组、(AM-MSCs+异氟烷)组,造模后6 h给予相应治疗。移植后7 d各组随机处死6只,采用RT-PCR、Western Blot检测脑组织中GAP-43、AQP4 mRNA和蛋白表达的变化。移植后24 h、3 d及造模后1、2、3、4周行动物神经功能缺损(NSS)评分,在移植后21~28 d进行Morris水迷宫试验。4周后处死行免疫组化、HE染色观察病理学变化。结果:移植后7 d,(AM-MSCs+异氟烷)组大鼠脑组织中GAP-43mRNA和蛋白表达高于脑梗死组、AM-MSCs组、异氟烷组(P0.05);异氟烷组和AM-MSCs组的大鼠脑组织中GAP-43 mRNA和蛋白表达水平高于脑梗死组(P0.05)。(AM-MSCs+异氟烷)组的AQP4 mRNA和蛋白表达低于其他各组(P0.05);AM-MSCs组和异氟烷组低于脑梗死组(P0.05)。移植后1周,(AM-MSCs+异氟烷)组的NSS评分低于异氟烷组、AM-MSCs移植组,异氟烷组、AM-MSCs移植组低于脑梗死组(P0.05)。移植后3~5 d,(AM-MSCs+异氟烷)组平均逃逸潜伏期较异氟烷组、AM-MSCs组、脑梗死组缩短(P0.05或P0.01)。(AM-MSCs+异氟烷)组的目标距离百分比高于异氟烷组和AM-MSCs组(P0.05),并高于脑梗死组(P0.01)。伤后4周,脑梗死组未见神经轴索通过,异氟烷组、AM-MSCs移植组可见少量神经轴索样结构,(AM-MSCs+异氟烷)组可见较多神经轴索样结构(P0.05)。CM-Dil的阳性细胞数(AM-MSCs+异氟烷)组高于AM-MSCs组,异氟烷组高于脑梗死组,假手术组最少(P0.05)。结论:异氟烷干预联合AMMSCs移植治疗大鼠脑梗死可明显改善大鼠的神经学功能。

Effect of Human Amnion Mesenchyme Stem Cell Transplantation Combined with Isoflurane on Rats with Cerebral Infarction

Objective:To observe the effect of amnion mesenchyme stem cells (AM-MSCs) transplantation combined with isoflurane on the recovery of neurological function in rats with cerebral infarction.Methods:One hundred rats model of cerebral infarction of middle cerebral artery occlusion was established by suture method,which were randomly divided into groups sham-operation,cerebral infarction,AM-MSCs,isoflurane and (AM-MSCs+isoflurane),corresponding treatment given at 6 h after modeling.Six rats in each group were sacrificed randomly after transplantation,and the changes of GAP-43,AQP4 mRNA and protein expression in brain tissue were detected by RT-PCR and Western Blot.After 24 h,neurological severity score (NSS) was performed at 1,2,3 and 4 weeks after transplantation.The Morris water maze test was performed at 21～28 d after transplantation.After 4 weeks,the rats were killed and the pathological changes were observed by immunohistochemistry and HE staining.Results:At 7 d after transplantation,GAP-43 mRNA and protein of (AM-MSCs+isoflurane) group in brain tissue were higher than those of groups cerebral infarction,AM-MSCs and isoflurane (P＜0.05).The expression of GAP-43 mRNA and protein levels in cerebral infarction group were higher than those of groups isoflurane and AM-MSCs (P＜0.05).The expression ofAQP4 mRNA and protein in (AM-MSCs+ isoflurane) group was lower than those in other groups (P＜0.05).AM-MSCs group and isoflurane group were lower than those in cerebral infarction group (P＜0.05).At 1 week after transplantation,the NSS score of the (AM-MSCs+isoflurane) group was lower than that of groups isoflurane and AM-MSCs,while that of isoflurane group and AM-MSCs transplantation group were lower than that of cerebral infarction group (P＜0.05).At 3～5 d after transplantation,the mean escape latency of (AM-MSCs+isoflurane) group was shorter than that of groups isoflurane,AM-MSCs and cerebral infarction (P＜0.05 or P＜0.01).The percentage of target distance of (AM-MSCs+isoflurane) group was higher than that of groups isoflurane and AM-MSCs (P＜0.05),and higher than that of cerebral infarction group (P＜0.01).At 4 weeks after injury,there was no axon in the cerebral infarction group,and there was a small amount of nerve axon like structure in the groups isoflurane and AM-MSCs,and there was a lot of nerve axon like structure (AM-MSCs+isoflurane) group (P＜0.05).The number of CM-Dil positive cells of (AM-MSCs+isoflurane) group was higher than that in AM-MSCs group,while that in isoflurane group was higher than that in cerebral infarction group (P＜0.05).Conclusion:Isoflurane intervention combined with AM-MSCs transplantation can improve neurological function in rats with cerebral infarction.