【作者】 易兵鸿；

【导师】 谢小平；

【作者基本信息】 南昌大学 ， 外科学， 2010， 硕士

【摘要】 目的:探讨5-氟脲嘧啶壳聚糖缓释微球对裸鼠腹腔种植胃癌细胞生长的抑制作用。方法:采用乳化化学交联法制备5-氟脲嘧啶壳聚糖缓释微球:将5-氟脲嘧啶溶于5%醋酸溶液,加入壳聚糖,配成水相;以石蜡油为油相;将水相滴入油相,超声乳化, 25%戊二醛交联固化,真空干燥得微球。同法,仅不加5-氟脲嘧啶制得空白壳聚糖微球。观察微球的基本性状,并测定5-氟脲嘧啶壳聚糖缓释微球的药物包封率及载药量。将微球142 mg(含5-氟脲嘧啶50mg)及5-氟脲嘧啶粉剂50mg分别置于透析袋中,每小时1次,24h后每天1次,共7d,测定各时段微球释药量及药物累计释放率。建立裸鼠胃癌细胞腹腔种植模型,按体表面积计算5-氟脲嘧啶用量,换算出5-氟脲嘧啶壳聚糖缓释微球用量,将祼鼠随机分为4组:A组(5-氟脲嘧啶壳聚糖缓释微球组)、B组(空白壳聚糖微球组)、C组(5-氟脲嘧啶组)、D组(生理盐水组),分别于裸鼠腹腔内注入5-氟脲嘧啶壳聚糖缓释微球、空白壳聚糖微球、5-氟脲嘧啶注射剂、生理盐水。全部裸鼠在SPF条件下饲养至1组裸鼠全部死亡。观察各组祼鼠的体重、腹围变化、腹腔癌结节计数、腹水胃癌细胞凋亡率、祼鼠生存率和药物毒副作用。结果:所制5-氟脲嘧啶壳聚糖缓释微球分布均匀,平均粒径为(185.5±15.0)nm,载药量为(35.2±0.8)%,药物包封率为(49.3±2.1)%。体外药物缓释实验显示,所制5-氟脲嘧啶壳聚糖缓释微球第1h 5-氟脲嘧啶释放量为21.4±2.0μg/ml,以后逐渐减少,第1d 5-氟脲嘧啶累计释放量为123.2±3.6μg/ml,以后呈持续缓慢释放,至第7d仍释放4.5±0.6μg /ml。在4组裸鼠中,A组裸鼠的腹围、体重在用药前后无明显改变(P >0.05),A组裸鼠的生存率明显高于B组裸鼠(P<0.01)、C组裸鼠(P<0.05)和D组裸鼠(P<0.01),并且,A组裸鼠腹腔癌结节数也较其余3组裸鼠明显减少(P <0.05),而且,腹水胃癌细胞凋亡率明显高于B组祼鼠(P<0.05)、C组祼鼠(P<0.05)和D组祼鼠(P<0.01);然而, A组裸鼠静脉血WBC总数、ALT水平、BUN水平、TBI水平与其他三组裸鼠比较无明显差异(P> 0.05)。结论:5-氟脲嘧啶壳聚糖缓释微球粒径分布均匀,具有较好的药物包封率及药物缓释性;用于胃癌腹腔化疗可明显抑制祼鼠腹腔种植胃癌细胞的生长,改善胃癌细胞腹腔种植祼鼠的预后。更多还原

【Abstract】 Objective: Investigation the growth inhibitory effection of 5- fluorouracil loaded chi- tosan sustained-release microspheres to the gastric cancer cells of peritoneal implantation in nude mice.Methods: The microspheres were prepared using emulsified-chemical cross-linking method : the water phase which consisted of 5-fluorouracil dissolved in 5% acetic acid and chitosan added after 5-fluorouracil was dissolved was droppded into the oil phase of paraffin oil , emulsification by ultrasound and using 25% glutaraldehyde to cross-link, then obtained the microspheres by vacuum drying. The same method, only without 5 - fluorouracil obtained blank microspheres. Observed basic characteristics of the microspheres and studied the drug encapsulation efficiency and the Drug load- ing. Weighed the microspheres 142 mg (include 5 - fluorouracil 50mg)and 5 - fluorouracil powder 50mg , placed them in dialysis bags, observed the role of drug release in vitro by determining con- centration and cumulative release hourly in 24h、then Once a day and a total of seven days . Establish the model of peritoneal implantation of gastric cancer cells in nude mice, calculated according to body surface area of 5 - fluorouracil dosage, con- version out of the dosage of 5- fluorouracil loaded chitosan sustained-release micro- spheres. All 32 nude mice divided into four groups randomly: A group(5- fluorouracil loaded chito- san sustained-release microspheres group)、B group(blank microspheres group)、C group(5- fluorouracil group)and D group(NS group), the nude mice were in- jected into the abdominal cavity 5- fluorouracil loaded chitosan sustained-release microspheres、blank microspheres、5- fluorouracil、NS by groups. Studied the changes of the every mouse body weight and abdominal circumference、the number of peritoneal metastatic noduli、the rate of gastric cancer cells apoptosis、survival of the nude mouse and the drug toxicity.Results: The average particle size of the microspheres was (185.5±15.0)nm, drug-loading rate was (35.2±0.8)% and encapsulation efficiency was ( 49.3±2.1)%. The microspheres release experiments in vitro showed that it released 5-fluorouracil 21.4±3.6μg/ml in the first hour、123.2±3.6μg/ml on the first day and 4.5±0.6/ml on the seventh day by releasing slowly. In the nude mice of A group ,The survival rate was more than the nude mice of B group (P< 0.05)、the nude mice of C group(P< 0.05)and the nude mice of D group(P< 0.01), the number of peritoneal metastatic noduli in the nude mice of A group was less than the nude mice of other group(P<0.05), the rate of gastric cancer cells apoptosis in the nude mice of A group was more than the nude mice of B group (P< 0.05)、the nude mice of C group(P<0.05)and the nude mice of D group(P< 0.01) when the difference in the number of WBC、ALT、BUN、TBI of Vein blood in the nude mice compared with other groups showed no statistical significance (P> 0.05).Conclusions: The 5- fluorouracil loaded chitosan sustained- release microspheres have uniform distribution、good drug encapsulation efficiency and drug release. The microspheres show perfect growth inhibition effect to gastric cancer cells of peritoneal implantation in nude mice and can improve the prognosis of the mice.更多还原