微囊化PC12细胞脑内移植治疗帕金森病的实验研究

Therapeutic effect of encapsulated-PC12 cells transplanted into the brain of experimental Parkinsonian rats

目的 观察微囊化 PC12细胞脑内移植对帕金森病 (PD)大鼠模型的治疗作用。方法 以 6 - OHDA分两点注入大鼠纹状体制备 PD模型。应用国产新型材料壳聚糖制成的海藻酸钠 -壳聚糖 -海藻酸钠 (ACA )微囊包裹PC12细胞 ,分别将微囊化 PC12细胞、裸 PC12细胞、空微囊移植入 PD模型鼠损伤侧纹状体内 ,以阿朴吗啡检测移植前后大鼠旋转行为的差异。行纹状体和黑质的 HE染色和酪氨酸羟化酶免疫组化染色观察其病理形态学的变化。对移植的微囊化 PC12细胞回收后再培养 ,以 MTT和台盼兰检测细胞活性。结果 微囊化 PC12细胞移植能够改善PD模型鼠的旋转行为 ,与移植前和空微囊移植组相比有显著差异 (P<0 .0 5 ) ,症状改善至少持续 3个月。裸 PC12细胞移植组大鼠的旋转行为也有改善 ,与移植前相比有统计学差异 (P<0 .0 5 ) ,但仅持续了 2个月 ,移植 8周后又渐回到移植前水平 ,且部分大鼠颅内有致死性肿瘤形成。空微囊移植组移植前后大鼠的旋转行为无明显差异 (P>0 .0 5 )。回收微胶囊内的 PC12细胞再培养生长良好 ,MTT法和台盼兰法检测显示细胞具有生物活性。结论 微囊化 PC12细胞脑内移植能够改善阿朴吗啡诱发的 PD模型鼠的旋转症状 ,ACA新型微包囊能够起到有效的免疫隔离和抑制肿瘤形成的作用。

Objective To observe the therapeutic effect of transplanting the encapsulated-PC12 cells into the brain of experimental Parkinsonian rats. Methods 6-OHDA was stereotaxicaly injected at two sites points the striatum of the rats in order to fabricate PD models. The Alginate-Chitosan-Alginate was used as the microcapsule. Encapsulated-PC12 cells、PC12 cells and empty microcapsule were transplanted into the striatum respectively ipsilateral to the lesion. The behavioral changes were tested by apomorphine. The morphological alterations were observed by striatal and substantia nigral HE stain and immunohistochemical stain of tyrosine hydroxylase. Reculture the retrieved encapsulated PC12 cells and test their activity by MTT and Trypan blue. Results The encapsulated PC12 cells can improve the rotational behavior of PD model rats and have a significant difference(P< 0.05) compared to pretransplantation and empty capsule transplantation group. This behavioral improvement can last at least three months. Although the unencapsulated PC12 cells can also improve the rotational behavior,the symptom improvement lasted only two months and returned to pretransplantation level at eight week after operation. In addition,several of the grafts changed into mortal tumor. There is no significant difference between before and after transplantation in empty capsule group(P> 0.05). The well growth and the MTT and Trypan blue test proved the good biological activity of the retrieved PC12 cells. Conclusion The transplantation of encapsulated-PC12 cells can improve the rotational behavior of PD model rats induced by apomorphine. ACA microcapsule can both isolate the host immune system effectively and prevent the hypersegmentation of the cells inside the capsule.