嗅鞘细胞联合神经干细胞脑内移植改善Parkinson病大鼠行为的观察

为了观察嗅鞘细胞(OECs)联合神经干细胞(NSCs)脑内移植对Parkinson病(PD)的治疗作用,首先将6-OHDA注射至左侧黑质致密部和腹侧被盖区制备PD大鼠。将成功制备的PD大鼠随机分为5组,即0.9%生理盐水对照组、单纯移植NSCs或OECs组、OECs+NSCs共移植组和PD模型对照组。将培养并传至第3代的NSCs、OECs和OECs+NSCs分别移植到PD大鼠的纹状体内,于移植后7、14、30、60d检测PD大鼠旋转行为变化后处死,取脑做冰冻切片,行BrdU、酪氨酸羟化酶(TH)、p75免疫荧光和TH免疫组织化学检测。移植后21d以内,各组大鼠的旋转行为无明显变化;30d和60d时,单纯移植NSCs或OECs组和OECs+NSCs共移植组与生理盐水和模型对照组相比,旋转行为有较明显的改善(P<0.05),其中OECs+NSCs共移植组的旋转行为改善更为显著。在移植后30d和60d,单纯移植NSCs或OECs组在移植区可见少数TH阳性神经元,而OECs+NSCs共移植组的TH阳性神经元明显多于单纯移植NSCs或OECs组(P<0.05)。上述结果表明:OECs能促进胚鼠中脑来源的NSCs向TH阳性神经元分化;单纯移植NSCs或OECs和OECs+NSCs共移植,均能不同程度地改善PD大鼠的旋转行为,但联合移植优于单纯移植。

OBSERVATION ON THE EFFECTS OF NEURAL STEM CELLS AND OLFACTORY ENSHEATHING CELLS INTRACEREBRAL CO-GRAFT ON THE BEHAVIOR CHANGES IN THE PARKINSON'S DISEASE RAT

To observe the effects of the neural stem cells(NSCs)and olfactory ensheathing cells (OECs) intracerebral co-graft on the behavior changes in the rat model of Parkinson’s disease (PD). The rat model of the PD was established with injecting 6-OHDA into the substantia nigra compacta part (SNc) and ventral tegmental area (VTA). The PD rats were divided into 5 groups: 0.9% normal saline (NS) control group, pure NSCs or OECs transplantation group, NSCs+OECs co-graft group and PD model control group. NSCs and OECs derived from rat embryonic midbrain and olfactory bulb were prepared by culture. The changes of rotating behavior were measured by comparing with different groups at different time points after transplantation. The survived cells were detected by BrdU, p75, TH/p75 immunofluorescent and TH immunohistochemical staining, respectively. Compared with control groups, there were no obvious changes during the first 3 weeks after transplantation. But at 30 d and 60 d, the rotation was distinctly improved (P<0.05) in the pure NSCs or OECs transplantation group and NSCs+OECs transplantation group, especially in the later group. The emergence of TH-positive cells was at 14 d after transplantation. TH-positive cells were gradually increased during 30 d and 60 d after transplantation. Comparing with NSCs or OECs group, there were more TH-positive cells in the NSCs+OECs group. Our data suggest that OECs may promote the NSCs derived from rat embryonic mesencephalon to become dopaminergic neurons after intracerebral transplantation.