VCAM-1、TLR2在亚急性1-甲基-4-苯基-1,2,3,6-四氢吡啶帕金森病小鼠模型纹状体中表达的研究

VCAM-1 and TLR2 in the caudate putamen of subacute MPTP Parkinson mouse model

目的研究亚急性1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)帕金森病小鼠模型纹状体中VCAM-1、TLR2的表达。方法腹腔注射MPTP制作亚急性(20mg/kg,每天1次,共7d)帕金森病小鼠模型。实验分为3组,8～10周龄小鼠组、8月龄中年小鼠组、亚急性小鼠模型组,每组6只C57BL/6小鼠。对照组小鼠(8～10周龄组、8月龄中年小鼠组)腹腔注射等量的生理盐水。模型组小鼠在末次注射MPTP 1d后取脑组织。用胶质细胞纤维酸性蛋白(GFAP)免疫组织化学染色,观察亚急性小鼠模型纹状体中炎性标志细胞——星型胶质细胞改变的情况。用RT-PCR方法检测VCAM-1 mRNA、TLR2 mRNA的基因表达水平。用Western blot方法检测VCAM-1蛋白表达水平。结果亚急性小鼠模型纹状体GFAP阳性细胞明显增多。亚急性模型组纹状体VCAM-1 mRNA表达水平明显增高,高于正常对照8～10周龄小鼠组和8月龄中年小鼠组(P<0.05),TLR2 mRNA表达各组间没有显著性差异(P>0.05)。亚急性模型组纹状体VCAM-1蛋白表达水平亦增高,高于正常对照组和中年小鼠组(P<0.05)。结论 VCAM-1在MPTP帕金森病小鼠纹状体中表达增高,可能与炎症反应有关,从而参与神经损伤和帕金森病病理生理过程。

Objective To investigate VCAM-1 and TLR2 in the caudate putamen of subacute MPTP Parkinson mouse model.Methods C57BL/6 mice were divided into 8～10 week-old mice group,middle-aged（8 month-old）mice group,and subacute MPTP parkinson model group.Model mice were intraperitoneally injected with MPTP（20mg/kg）once daily for consecutive 7 days,and were sacrificed 1 day after the last injection.Control mice（8～10 week-old mice group,middle-aged mice group）received saline solution only.Astrocyte of subacute mice model was observed by using GFAP stain.The gene expression of VCAM-1 mRNA and TLR2 mRNA in those brain samples were analyzed by RT-PCR.The protein expression of VCAM-1 was measured by western blot.Results The number of GFAP positive cells increased after MPTP treatment.The expression of VCAM-1 mRNA and VCAM-1 protein but not TLR2 mRNA increased in the caudate putamen of mice treated with MPTP（subacute Parkinson model）in comparison to untreated animals.Conclusion The overexpression of VCAM-1 suggest that VCAM-1 may be associated with inflammatory response in the pathophysiology of Parkinson disease.