姜黄素炎症靶向自微乳的制备与评价研究

发布时间：2018-06-21 00:26

本文选题：姜黄素 + 负电荷　；参考：《北京中医药大学》2017年硕士论文

【摘要】：目的:利用结肠炎症部位阳离子蛋白表达增高,结肠炎症部位带正电的特点,构建负电荷的姜黄素炎症靶向自微乳给药系统(NC-CUR-SMEDDS)。对自微乳进行初步质量评价,并对自微乳的体外靶向性和体内药效学进行探讨,为炎症靶向制剂的开发提供新思路,为实现溃疡性结肠炎的炎症靶向治疗提供理论和数据基础。方法:在前期姜黄素自微乳(CUR-SMEDDS)的研究基础上,以姜黄素为模型药物,以乳剂的粒径、Zeta电位、包封率和载药量为评价指标,通过单因素试验筛选电荷调节剂的最佳用量,制备NC-CUR-SMEDDS。通过观察微乳外观和微观形态并测定其粒径、Zeta电位、包封率及载药量对其进行质量评价。并采用5%葡聚糖硫酸钠(DSS)造成BALB/c小鼠急性溃疡性结肠炎模型,通过间接体内(ex vivo)实验、小动物在体荧光检测技术考察自微乳在炎症部位的黏附情况,评价其体外炎症靶向效果。同时,采用保留灌肠的给药方式,将药物直接靶向于结肠部位,以5-氨基水杨酸(5-ASA)为阳性对照药,非炎症靶向自微乳(CUR-SMEDDS)作为对照组,以疾病活动指数评分(DAI)、结肠长度、结肠黏膜组织病理学评分、结肠组织中丙二醛(MDA)含量、髓过氧化物酶(MPO)活性、白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)的含量为评价指标,评价NC-CUR-SMEDDS体内靶向治疗效果。结果:以丁二酸二辛酯磺酸钠为最佳电荷调节剂,单因素实验筛选得到NC-CUR-SMEDDS的最优处方:聚氧乙烯氢化蓖麻油RH40(CremophorRH40):二乙二醇单乙基醚HP(Transcutol HP):丙二醇单辛酸酯90(Capryol 90):丁二酸二辛酯磺酸钠(Docusate Sodium)=6:3:1:0.4,所形成的微乳 Zeta 电位可以达到-43.43±0.29mV,外观澄清、透明,粒径分布均匀,平均粒径为(14.08±0.082)nm,姜黄素载药量为37.98 mg·g-1,包封率为99.22%。间接体内(ex vivo)黏附实验结果表明,NC-CUR-SMEDDS(Zeta电位为-43.43±0.29mV)在DSS诱导小鼠UC模型结肠炎症部位体现出良好的炎症靶向作用,荧光半定量结果显示,NC-CUR-SMEDDS在炎症部位的吸附是正常组织部位的2.8倍,具有显著性差异(P0.0001);同时,NC-CUR-SMEDDS与CUR-SMEDDS(Zeta电位为-6.28±0.86mV)相比,NC-CUR-SMEDDS在炎症部位吸附较多,是CUR-SMEDDS的1.5倍(P0.01),体现出很好的炎症靶向作用。通过保留灌肠结肠定向给药,NC-CUR-SMEDDS对DSS诱导的小鼠UC模型具有明显的改善作用,能够减轻小鼠的临床症状,包括精神活动状态、便血、体重等。同时,NC-CUR-SMEDDS对于小鼠结肠组织充血、水肿、溃疡等症状具有不同程度的缓解作用,病理组织学检查结果显示,NC-CUR-SMEDDS组炎症细胞浸润减少,隐窝结构相对完整,杯状细胞存在较多。生化指标检测结果显示NC-CUR-SMEDDS显著降低小鼠结肠组织中MPO的活性以及MDA、TNF-α和IL-6的含量,治疗作用与阳性对照组接近。结论:本课题通过改变粒子表面电性,构建的姜黄素负电荷自微乳给药系统包封率高,粒径分布均匀,实现了姜黄素自微乳在结肠炎症部位的特异性靶向,增加姜黄素在溃疡性结肠炎炎性细胞表面的药物浓度,为炎症靶向制剂系统提供了新思路和新方法。
[Abstract]:Objective: using increased colitis site cationic protein expression, characteristics of colitis parts of positive, negative charge of construction inflammation targeted curcumin self microemulsifying drug delivery system (NC-CUR-SMEDDS). The preliminary evaluation of the quality of self microemulsion, and the in vitro self microemulsion and pharmacodynamics in vivo to explore, for targeting inflammation preparation The development of new ideas, and to provide theoretical foundation for the realization of the target data of inflammation of ulcerative colitis treatment. Methods: in the early stage of curcumin self microemulsion (CUR-SMEDDS) based on the research, using curcumin as a model drug, the emulsion particle size, Zeta potential, entrapment efficiency and drug loading as evaluation index, adjusted by the single factor screening test charge The best dosage of agent, the preparation of NC-CUR-SMEDDS. by observing the appearance and microstructure of microemulsion and determine the particle size, Zeta potential, encapsulation efficiency and drug loading to evaluate its quality. And the use of 5% dextran sulfate sodium (DSS) caused by acute ulcerative colitis in mice model of BALB/c (ex, vivo) by ex vivo experiments, small animal in vivo fluorescence detection Investigation of self microemulsion at sites of inflammation adhesion, inflammation and evaluate its in vitro targeting effect. At the same time, the retention enema administration, the drug directly targeted to the colon, with 5- amino salicylic acid (5-ASA) as a positive control medicine, non inflammatory target self microemulsion (CUR-SMEDDS) as the control group, the disease activity index (DAI),. The length of intestine, colon mucosa pathological study score, malondialdehyde (MDA) content in colon tissue, myeloperoxidase (MPO) activity of interleukin -6 (IL-6) and tumor necrosis factor alpha (TNF- alpha) content as the evaluation index, evaluation of NC-CUR-SMEDDS targeting therapy in vivo. Results: two succinic acid ester is the best sodium charge regulator, single The experimental factors screened the optimal prescription of NC-CUR-SMEDDS: Cremophor RH40 (CremophorRH40): diethylene glycol monoethyl ether HP (Transcutol HP): propylene glycol monocaprylate 90 (Capryol 90) two: succinic acid octyl sulfonic acid sodium (Docusate Sodium) =6:3:1: 0.4, Zeta potential microemulsion formed can reach -43.43 + 0.29mV. The appearance of Weicheng Clear, transparent, uniform particle size distribution and average particle size (14.08 + 0.082) nm, curcumin loading was 37.98 Mg - g-1, the encapsulation rate of 99.22%. (ex vivo) ex vivo adhesion experiment results show that NC-CUR-SMEDDS (Zeta potential of -43.43 + 0.29mV) in DSS induced colitis in mice UC model of parts good inflammation targeting, semi quantitative fluorescence The amount of results showed that the adsorption of NC-CUR-SMEDDS at the site of inflammation is 2.8 times of normal tissues, with a significant difference (P0.0001); at the same time, NC-CUR-SMEDDS and CUR-SMEDDS (Zeta potential of -6.28 + 0.86mV) compared to NC-CUR-SMEDDS at the site of inflammation absorb more, is 1.5 times that of CUR-SMEDDS (P0.01), reflecting the good target of inflammation through to. Retention enema colon targeted delivery, NC-CUR-SMEDDS has obvious effect on mouse UC model induced by DSS, can alleviate clinical symptoms in mice, including mental activity, stool, weight and so on. At the same time, NC-CUR-SMEDDS in colon tissue of mice with hyperemia, edema, ulcers and other symptoms have different degrees of ease, histopathology Test results show that the decrease of NC-CUR-SMEDDS group inflammatory cell infiltration and crypt structure is relatively complete, there are many goblet cells. Biochemical test results showed that the NC-CUR-SMEDDS decreased significantly in mice colonic tissue MPO activity and MDA content of TNF- alpha and IL-6, the treatment effect and the positive control group. Conclusion: according to the change of particle Sub surface electricity, negative charge of curcumin self microemulsifying drug delivery system with high encapsulation efficiency, uniform particle size distribution, the specific target of curcumin self microemulsion in colitis parts to increase drug concentration, the effect of curcumin on ulcerative colonic inflammatory cell surface, inflammatory target provides new ideas and new methods to the preparation system.
【学位授予单位】：北京中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R283.6

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