摘要
目的：探讨多奈哌齐对实验性自身免疫性脑脊髓炎（EAE）小鼠模型血脑屏障渗透性的影响。方法将48只6-8周龄雌性C57BL／6小鼠分为多奈哌齐组（19只）、PBS 组（19只）与空白对照组（10只）。将多奈哌齐组及PBS组制备为EAE小鼠模型，在接种免疫后第13日，将多奈哌齐2 mg／（kg·d）溶解于PBS中对多奈哌齐组进行灌胃；而PBS组只予PBS进行灌胃。对EAE小鼠进行临床症状评分和组织学评估（炎症评分及脱髓鞘评分），利用伊文思蓝对EAE小鼠脑组织的蓝染程度评估血脑屏障渗透性，用蛋白免疫印迹法检测EAE小鼠脑组织中基质金属蛋白酶-2（MMP-2）和基质金属蛋白酶-9（MMP-9）的表达情况。结果多奈哌齐组的临床症状评分低于PBS组的临床症状评分（t＝4.351，P＜0.001）。多奈哌齐组的炎症评分低于PBS组（t＝6.325，P＜0.001）；PBS组的腰髓有大片的髓鞘脱失，但多奈哌齐组脱髓鞘病变较其少，2组评分比较差异有统计学意义（t＝3.308，P＝0.008）。与空白对照组相比，PBS组（t＝8.621，P＜0.001）和多奈哌齐组（t＝9.974，P＜0.001）脑内伊文思蓝含量均较高；与PBS组相比，多奈哌齐组脑内伊文思蓝含量较低（t＝2.286， P＝0.045）。与PBS组相比，多奈哌齐可抑制MMP-2（t＝4.679，P＝0.010）和MMP-9（t＝5.822，P＝0.001）的表达。结论多奈哌齐可降低EAE小鼠血脑屏障的渗透性。
Objective To evaluate the effect of donepezil upon inhibiting the blood-brain barrier permeability in mouse models with experimental autoimmune encephalomyelitis （EAE）. Methods Forty eight female C57BL/6 mice,aged 6-8 weeks,were divided into the donepezil （n =19 ）,phosphate buffer saline （PBS,n=19）and control groups （n=10）. EAE mouse models were established in the donepezil and PBS groups. In the donepezil group,intragastric gavage of PBS containing 2 mg/（kg·d）of donepezil was delivered at 13 d after innoculation,whereas PBS alone was administered in the PBS group. The grading of clinical symptoms and histological evaluation were performed in EAE mice （inflammation and demyelinating scores）.The blood-brain barrier permeability was assessed by Evans blue staining. The expression levels of matrix metalloproteinase-2 （MMP-2）and MMP-9 were measured by western blot. Results The score of clinical symp-toms in the donepezil group was significantly lower compared with that in the PBS group （t =4.35 1 ,P〈0.001）. The inflammation score in the donepezil group was considerably lower than that in the PBS group （t=6.325,P〈0.001）. In the PBS group,a large mass of myelinoclasis was observed,whereas demyelinating lesion was seldom seen in the donepezil group with statistical significance （t=3.308,P=0.008）. Compared with the control group,intracerebral content of Evans blue in the PBS （t=8.621,P〈0.001）and donepezil groups （t=9.974,P〈0.001）was significantly higher. However,intracerebral content of Evans blue in the donepezil group was significantly less compared with that in the PBS group （t=2.286,P=0.045）. Compared＆with the PBS group,donepezil could significantly down-regulate the expression levels of MMP-2 （t=4.679,P=0.010）and MMP-9 （t=5.822,P=0.001 ）. Conclusion Donepezil can reduce the blood-brain barrier permeability in EAE mouse models.
出处
《新医学》
2016年第12期803-808,共6页
Journal of New Medicine
基金
广东省医学科研基金(A2014233)
