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目的研究微囊化后的猪视网膜色素上皮细胞(retinal pigment epithelial,RPE)对帕金森病大鼠模型的移植疗效。方法原代培养RPE 并传代,高效液相色谱法测定培养液上清中多巴胺(dopamine, DA)和高香草酸(homovanillic acid, HVA)的含量,ELISA法检测脑源性神经营养因子(brain-derived neurotrophic factor, BDNF)和胶质细胞源性神经营养因子(glial-derived neurotrophic factor, GDNF)的含量。用高压静电成囊装置制备海藻酸钠-多聚赖氨酸-海藻酸钠微囊化细胞。6-羟基多巴胺(6-hydroxydopamine, 6-OHDA)毁损内侧前脑束 (medial fore-brain bundle,MFB)建立 SD 大鼠帕金森病模型。立体定向移植 RPE+ 微囊,检验旋转实验、免疫组化和脑内生化的变化。结果 RPE 培养上清液中DA、HVA、BDNF、GDNF 的含量稳定,微囊化后细胞长期存活,活性没有明显变化。6-OHDA毁损MFB建立大鼠帕金森病模型的成模率为83%。移植微囊化的RPE后有效率为33%。结论猪 RPE 体外培养生长旺盛,持续分泌 DA、BDNF 和 GNDF,微囊化不影响其分泌功能。RPE 移植对帕金森病大鼠有一定的治疗作用。
Objective To study the effect of microencapsulated pig retinal pigment epithelial (RPE) transplantation on Parkinson’s disease rat model. Methods RPE was cultured in primary culture and passaged. The content of dopamine (DA) and homovanillic acid (HVA) in culture supernatant was determined by high performance liquid chromatography (HPLC), and the expression of brain-derived neurotrophic factor (brain- derived neurotrophic factor (BDNF) and glial-derived neurotrophic factor (GDNF). Preparation of sodium alginate - polylysine - alginate microencapsulated cells with high voltage electrostatic capsule device. A rat model of Parkinson’s disease was established by 6-hydroxydopamine (6-OHDA) destroying the medial fore-brain bundle (MFB). Stereotactic RPE + microcapsules were transplanted and examined for rotation, immunohistochemistry and changes in brain biochemistry. Results The contents of DA, HVA, BDNF and GDNF in RPE culture supernatant were stable. The cells survived after microencapsulation and their activities did not change significantly. 6-OHDA damage MFB rat model of Parkinson’s disease was 83%. The efficiency of transplanted microencapsulated RPE was 33%. Conclusion Pig RPE grew vigorously in vitro and continued to secrete DA, BDNF and GNDF. Microencapsulation did not affect its secretion function. RPE transplantation on Parkinson’s disease rats have a certain therapeutic effect.