脑内移植治疗帕金森病的移植神经元凋亡时程的研究

为了探讨将 E14腹侧中脑 ( VM)细胞悬液植入 Parkinson病大鼠纹状体后的移植神经元的凋亡时程及其数量的变化规律 ,将实验动物分为移植组和空白对照组 ,两组均包括移植后 1、4、7和 2 8d四个时间点。分别运用 TU NEL 法、TH免疫组化法、H-E染色及透射电镜观察了凋亡细胞的生化和形态特征 ,藉以估计其凋亡细胞的数量变化状况。结果表明 ,移植组中用 TU NEL法观察移植神经元具有 TU NEL 阳性反应胞核的细胞在移植后 1、4、7、2 8d所占的百分比分别为 0 .14 9%、0 .13 1%、0 .0 66%和0 .0 0 5 % ,移植后 1、4d与 7、2 8d之间的比较差异显著 ( P<0 .0 0 1)。 TH免疫组化反应显示 ,在移植后 1、4、7、2 8d TH免疫反应阳性细胞存活率分别为 15 .7%、6.3 %、5 .9%和 4.9% ,移植后 1d与 4、7、2 8d之间的比较差异显著 ( P<0 .0 0 1)。本研究结果提示 :( 1)移植的胚 VM细胞凋亡主要见于移植后的头 7d,这一期间内大约有 90 %移植神经元发生凋亡 ;( 2 )这种凋亡对移植的TH细胞存活率的影响仅见于移植后的 4d。

A STUDY ON TIME COURSE OF APOPTOTIC CELL DEATH FOR GRAFT NEURONS TO THE BRAIN OF RAT MODEL PARKINSON'S DISEASE

In order to investigate the temporal pattern and magnitude of apoptotic cell death following the implantation of embryonic day 14(E14) ventral mesencephalic cell suspension grafts to DA denervated striatum, PD models were selected and divided randomly into two groups (A and B). Group A was graft group, and Group B was control group. The two groups were divided respectively into four time points at 1, 4, 7 and 28 days following grafting. All of the rats were killed at 1, 4, 7 and 28 days respectively. TUNEL technique, TH immunohistochemistry, H E staining and transmission electron microscope were used to assess the apoptotic cell death by detection of its biochemical and morphological identifiers respectively. In Group A, quantitative evaluations made of TUNEL positive nuclear profiles revealed significant differences between TUNEL positive nuclei at 1, 4 days and 7, 28 days after grafting (P<0.001), the average percentage of TUNEL positive nuclei was 0.149% at 1 day, 0.131% at 4 days,0.066% at 7 days, and 0.005% at 28 days postimplantation. Quantitative evaluations made of THir cells revealed significant differences between 1 and 4, 7 and 28 days after grafting(P<0.001). The average percentage of THir cells was 15.7% at 1 day, 6.3% at 4 days, 5.9% at 7 days and 4.9% at 28 days postimplantation. The results indicate: (1) The overwhelming majority of apoptotic cell death (90%) occurs within the first 7 days after transplantation. (2) The impact of the apoptosis that occur over the first week following grafting only appears to limit grafted THir neurons survival during the first 4 days. No significant differences between the survival rates of THir neurons at 4 and 28 days after grafting were found.