CAJ | 학술논문

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 forebrain 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.
목적 연구미낭화후적저시망막색소상피세포(retinal pigment epithelial,RPE)대파금삼병대서모형적이식료효.방법 원대배양RPE병전대,고효액상색보법측정배양액상청중다파알(dopamine,DA)화고향초산(homovanillic acid,HVA)적함량,ELISA법검측뇌원성신경영양인자(brain-derived neurotrophic factor,BDNF)화효질세포원성신경영양인자(glial-derived neurotrophic factor,GDNF)적함량.용고압정전성낭장치제비해조산납-다취뢰안산-해조산납미낭화세포.6-간기다파알(6-hydroxydopamine,6-OHDA)훼손내측전뇌속(medial forebrain bundle,MFB)건립SD대서파금삼병모형.입체정향이식RPE+미낭,검험선전실험、면역조화화뇌내생화적변화.결과 RPE배양상청액중DA、HVA、BDNF、GDNF적함량은정,미낭화후세포장기존활,활성몰유명현변화.6-OHDA훼손MFB건립대서파금삼병모형적성모솔위83%.이식미낭화적RPE후유효솔위33%.결론 저RPE체외배양생장왕성,지속분비DA、BDNF화GNDF,미낭화불영향기분비공능.RPE이식대파금삼병대서유일정적치료작용.
Object To investigate the therapeutic effect of microencapsulated porcine retinal pigmented epithelial cells (RPE-M) transplantation on rat model of Parkinson's disease (PD). Methods Primary porcine RPE cells were harvested by enzyme digestion and expanded in culture medium. Determine the levels of dopamine (DA) and homovanillic acid (HVA) by high performance liquid chromatography electrochemical (HPLC) assay, and the levels of brain-derived neurotrophic factor (BDNF) and glial-derived neurotrophic factor (GDNF) were detected by ELISA. Alginate-polylysine-alginate (APA)microencapsulated cells were produced by using a high voltage electrostatic system. PD rat model was established by unilateral injection of 6-hydroxydopamine (6-OHDA) into the medial forebrain bundle (MFB). After that, the RPE-M was transplanted into the corpus striatum of PD rat, and then the rotation test scores were recorded and biochemical changes of the corpus striatum were tested. Results The levels of DA, HVA, BDNF and GDNF secreted by RPE were stable in the RPE culture supernatant and were not changed by the microencapsulation. Eighty-three percent rats developed PD by unilateral lesion of 6-OHDA in the MFB. The RPE-M transplantation had therapeutic effect on 33% PD rats. Conclusion Porcine RPE cells grow actively in vitro and could secrete DA, HVA, BDNF, and GDNF constantly, which does not be affected by the passage culture and the APA miroencapsulation. RPE-M transplantation of may be a curative therapy for PD.