【摘要】：目的:觀察移植pIRESneo-EGFP-BDNF修飾骨髓間充質(zhì)干細胞(mesenchymal stem cells,MSCs)對帕金森病(Parkinson’sdisease,PD)大鼠的研究。方法 :將pEGFP(N1)-BDNF質(zhì)粒進行改造與pIRESneo相連結(jié),構(gòu)建高拷貝質(zhì)粒pIRESneo-EGFP-BDNF,采用電穿孔法轉(zhuǎn)染骨髓MSCs;制備PD大鼠模型,隨機分為假手術(shù)組、PD組、MSCs組、腦源性神經(jīng)生長因子(brain-derivedneurotrophic factor,BDNF)組,經(jīng)側(cè)腦室移植MSCs或pIRESneo-EGFP-BDNF修飾骨髓MSCs術(shù)后2、4、8周,腹腔注射阿樸嗎啡誘導(dǎo)PD大鼠旋轉(zhuǎn)行為;采用Western blot方法測定中腦黑質(zhì)BDNF、酪氨酸羥化酶(tyrosine hydroxylase,TH)蛋白的含量。結(jié)果:雙酶切鑒定pIRESneo-EGFP-BDNF構(gòu)建成功;移植術(shù)后2、4、8周MSCs組和BDNF組大鼠與PD組比較旋轉(zhuǎn)次數(shù)明顯減少(P0.05),以BDNF組較MSCs比較改善更為明顯(P0.05);移植細胞干預(yù)PD模型8周后中腦黑質(zhì)BDNF、TH蛋白的含量,BDNF組較MSCs組、PD組明顯增高(P0.05)。結(jié)論:經(jīng)側(cè)腦室移植pIRESneo-EGFP-BDNF修飾的骨髓MSCs能明顯改善PD大鼠的行為能力,顯著提高中腦黑質(zhì)BDNF、TH蛋白的含量。
[Abstract]:Aim: to observe the effects of pIRESneo-EGFP-BDNF modified bone marrow mesenchymal stem cells (mesenchymal stem cells,MSCs) on Parkinson's disease (Parkinson'sdisease,PD) rats. Methods: pEGFP (N1)-BDNF plasmid was modified with pIRESneo to construct the PD rat model of high copy plasmid pIRESneo-EGFP-BDNF, transfected with bone marrow MSCs; by electroporation. PD rats were randomly divided into sham-operation group, PD group, MSCs group, and brain derived nerve growth factor (brain-derivedneurotrophic factor,BDNF) group. The rotation behavior of PD rats induced by apomorphine was induced by intraperitoneal injection of apomorphine and the content of BDNF, tyrosine hydroxylase (tyrosine hydroxylase,TH) in the substantia nigra was measured by Western blot method. Results: pIRESneo-EGFP-BDNF was successfully constructed by double enzyme digestion. The rotation times of MSCs group and BDNF group were significantly decreased than that of PD group (P0.05), especially in BDNF group compared with MSCs group (P0.05). The content of BDNF,TH protein in substantia nigra was significantly increased in BDNF group than in MSCs group after 8 weeks of transplantation (P0.05). Conclusion: pIRESneo-EGFP-BDNF modified bone marrow MSCs can significantly improve the behavioral ability of PD rats and increase the content of BDNF,TH protein in the substantia nigra.
【作者單位】： 河北大學(xué)生物技術(shù)研究中心;河北大學(xué)附屬醫(yī)院神經(jīng)內(nèi)科;
【基金】：河北省科學(xué)技術(shù)研究與發(fā)展計劃資助項目(編號:08206120D)
【分類號】：R-332

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