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背景:细胞组织的微囊化移植是近年来帕金森病治疗的研究热点之一,目前发展较成熟、应用较多的是海藻酸钠-聚赖氨酸-海藻酸钠微胶囊,但其易于囊周纤维化、易破碎等缺点,使其临床应用受到了限制。应用国内研制的新型微胶囊材料海藻酸钠-壳聚糖-海藻酸钠微囊将PC12细胞微囊化后移植入帕金森病模型鼠纹状体内,观察其作用。目的:观察海藻酸钠-壳聚糖-海藻酸钠微囊化PC12细胞脑内移植治疗,改善帕金森病模型大鼠旋转行为的作用。设计:随机对照动物实验。单位:吉林大学第二医院和中国科学院大连化学物理研究所。材料:成年雄性Wistar大鼠40只,体质量(220±10)g;海藻酸钠-壳聚糖-海藻酸钠微囊;PC12细胞。方法:实验于2002-05/12在吉林大学第二医院动物实验室和中国科学院大连化学物理研究所完成。①应用国产新型材料壳聚糖制成的海藻酸钠-壳聚糖-海藻酸钠微囊化PC12细胞。②将成功建立的23只帕金森病大鼠模型随机分为3组,微囊化PC12细胞组10只、裸PC12细胞组7只、空微胶囊组6只。分别将海藻酸钠-壳聚糖-海藻酸钠微囊化PC12细胞、裸PC12细胞、空微胶囊移植入帕金森病模型鼠损伤侧纹状体内。③以阿朴吗啡检测移植前后大鼠旋转行为的差异。观察黑质及纹状体内微包囊的形态并检测微胶囊内细胞的活性。主要观察指标:①移植前后大鼠的旋转行为。②黑质和纹状体的病理形态。③回收的微包囊的完整性及囊内PC12细胞的活性。结果:①微囊化PC12细胞移植组在移植4周时旋转行为显著低于移植前和空微囊移植组犤(6.9±2.8),(11.7±5.5),(10.5±1.6)r/min,P<0.05犦,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与移植前相比也有改善犤(5.6±1.1),(9.5±1.5)r/min,P<0.05犦,但仅持续了2个月,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。②回收微胶囊内的PC12细胞再培养生长良好,并且具有生物活性。结论:微囊化PC12细胞脑内移植能够改善阿朴吗啡诱发的帕金森病模型鼠的旋转症状,海藻酸钠-壳聚糖-海藻酸钠新型微胶囊具有免疫隔离和抑制肿瘤形成的作用,有着广泛的临床应用前景。
BACKGROUND: Microencapsulated transplantation of cells and tissues is one of the research hotspots in the treatment of Parkinson’s disease in recent years. At present, the more mature and more used sodium alginate-polylysine-alginate microcapsules, Peribulbar fibrosis, easy to break and other shortcomings, its clinical application has been limited. The PC12 cells were microencapsulated and transplanted into the striatum of Parkinson’s disease model mice to observe the effect of sodium alginate-chitosan-alginate microcapsules. OBJECTIVE: To observe the effect of sodium alginate-chitosan-alginate microencapsulated PC12 cells intracerebral transplantation in improving the rotational behavior of rats with Parkinson’s disease. Design: Randomized controlled animal experiments. Unit: Second Hospital of Jilin University and Dalian Institute of Chemical Physics, Chinese Academy of Sciences. MATERIALS: Forty adult male Wistar rats with a body weight of (220 ± 10) g, sodium alginate-chitosan-alginate microcapsules and PC12 cells were obtained. METHODS: The experiment was performed at Animal Laboratory of the Second Hospital of Jilin University and Dalian Institute of Chemical Physics, Chinese Academy of Sciences from May 2002 to December 2002. ① using domestic new material made of chitosan alginate - chitosan - alginate microencapsulated PC12 cells. ② The successful establishment of 23 rat models of Parkinson’s disease were randomly divided into three groups, 10 microencapsulated PC12 cells, 7 PC12 bare cells and 6 empty microcapsules. The alginate-chitosan-alginate microencapsulated PC12 cells, naked PC12 cells and empty microcapsules were respectively transplanted into injured striatum of Parkinson’s disease model rats. (3) Apomorphine was used to detect the difference of rat rotation behavior before and after transplantation. The morphology of microcapsules in the substantia nigra and striatum was observed and the activity of cells in the microcapsules was examined. MAIN OUTCOME MEASURES: ① Rotation behavior of rats before and after transplantation. ② substantia nigra and striatum pathological morphology. ③ the integrity of the recovered microcapsules and the activity of PC12 cells in the capsule. RESULTS: ① The rotational behavior of microencapsulated PC12 cells at 4 weeks after transplantation was significantly lower than that before transplantation (6.9 ± 2.8), (11.7 ± 5.5), (10.5 ± 1.6) r / min, P <0.05 犦 and symptom improvement for at least 3 months. The rotation behavior of rats in PC12 cell transplantation group was also significantly improved (P <0.05 犦, P <0.05 犤) compared with those before transplantation , But lasted for only 2 months, and some of the rats had intracerebral fatal tumor formation. The rotation behavior of rats in empty microcapsule transplantation group had no significant difference before and after transplantation. ② PC12 cells in the microcapsules were recovered and cultured well and had biological activity. CONCLUSION: Transplantation of microencapsulated PC12 cells can improve apomorphine-induced Parkinson’s disease in mice. The new type of sodium alginate-chitosan-alginate microcapsules has immunosuppression and inhibition of tumor formation. Has a wide range of clinical applications.