移植BDNF修饰骨髓间充质干细胞对帕金森病模型大鼠TH表达的影响

The influence on expression of tyrosine hydroxylase after rats with Parkinson ' s disease modal treated by bone mesenchymal stem cell modified by plasmid pIRESneo-EGFP-BDNF

目的:观察移植pIRESneo-EGFP-BDNF修饰骨髓间充质干细胞(MSCs)对帕金森病模型大鼠TH表达的影响。方法:采用电穿孔法将pIRESneo-EGFP-BDNF转染至骨髓MSCs;通过脑内注射6-OHDA制备PD大鼠模型,随机分为Sham组,PD组,MSCs组,BDNF组,经侧脑室移植MSCs或pIRESneo-EGFP-BDNF修饰骨髓MSCs,术后2、4、8周,腹腔注射阿朴吗啡诱导PD大鼠旋转行为,移植BDNF修饰骨髓MSCs在大鼠脑组织内的迁移,采用免疫组化及Western blot方法测定各组大鼠中脑黑质TH蛋白表达的变化。结果:移植术后2、4、8周MSCs组和BDNF组大鼠与PD组比较旋转次数明显减少(P<0.05),以BDNF组较MSCs组比较改善更为明显(P<0.05),移植细胞干预PD模型大鼠8周后中脑黑质TH表达,BDNF组较MSCs组、PD组明显增高(P<0.05),移植pIRESneo-EGFP-BDNF修饰骨髓MSCs可以在PD大鼠脑内存活,并迁移至远处。结论:经侧脑室移植pIRESneo-EGFP-BDNF修饰的骨髓MSCs通过提高中脑黑质TH的活性,能够明显改善PD模型大鼠的行为能力。

Objective： To observe influence on expression of tyrosine hydroxylase （TH） after modal rats with Parkinson ＇s disease （PD） treated by bone mesenchymal stem ceils （MSCs） modified by plasmid pIRESneo-EGFP-BDNF. Meth- ods ： pIRESneo-EGFP-BDNF was transfected to MSCs with electroproation. The rats of Parkinson＇s disease model were set up by 6-OHDA injected into the brain and then divided into four groups randomly, which were Sham group, PD group, MSCs group, BDNF group. The rotating behavior of model rats induced by apomorphine intraperitoneally which transplan- ting bone MSCs or MSCs modified by plasmid pIRESneo-EGFP-BDNF through cerebral lateral ventricle after 2, 4 and 8 weeks. The expression of TH proteins in mesencephalic nigra were measured by Western blot and immunohistochemistry. Results： The rotation numbers of MSCs group or BDNF group in the 2, 4 and 8 weeks after transplanting were significant decrease compared with PD group （ P 〈 0.05 ）. Those of BDNF group were especially significant compared with MSCs group （ P 〈 0.05 ）. The expressional levels of TH protein in nigra after PD model rats intervented by cell transplantation 8 weeks were increased significantly in BDNF group, while compared with MSCs group or PD group respectively （P 〈 0.05）. The transferred bone MSCs modified by BDNF showed excellent survival rates in PD rat cerebral and distant mi- gration was observed. Conclusion： The behavior of modal rats with Pakinson＇s disease was improved significantly by in- creasing the level of TH in mesencephalic nigra while transplanting BDNF modified bone MSCs through cerebral lateral ventricle.