本文關(guān)鍵詞： 聚唾液酸 膠束 類(lèi)風(fēng)濕性關(guān)節(jié)炎 地塞米松 葉酸 膽固醇　出處：《鄭州大學(xué)》2017年碩士論文　論文類(lèi)型：學(xué)位論文

【摘要】：類(lèi)風(fēng)濕性關(guān)節(jié)炎是一種慢性、免疫系統(tǒng)紊亂性疾病,發(fā)病率約占人群的1%。藥物的給藥系統(tǒng)有效的解決了許多藥物低效和高毒的缺點(diǎn)。納米尺寸的藥物給藥系統(tǒng)由于能夠選擇性的聚集在病變部位而成為目前研究的熱點(diǎn),天然多糖是無(wú)毒、可生物降解的大分子生物材料,可用于制備納米尺寸的藥物載體。聚唾液酸(PSA)是具有強(qiáng)親水性的多糖,可以減少網(wǎng)狀內(nèi)皮系統(tǒng)對(duì)納米載體的攝取并延長(zhǎng)藥物在體內(nèi)的循環(huán)時(shí)間。膽固醇是細(xì)胞膜的一個(gè)組成部分,有利于納米載體的內(nèi)吞作用,充當(dāng)疏水基團(tuán)。本研究設(shè)計(jì)和合成了基于PSA-CC的納米混合膠束,用于改善抗炎藥物的遞送,提高藥效和減小毒性。通過(guò)NMR,IR,Size,ζ電位和透射電子顯微鏡證實(shí)了混合膠束合成成功�；赑SA的普通膠束和靶向混合膠束通過(guò)自組裝形成膠束的臨界膠束濃度分別為46.2±3.9μg/ml、32.1±5.2μg/ml。經(jīng)典抗炎藥物地塞米松通過(guò)疏水相互作用有效地加載到膠束中,地塞米松的載入使靶向混合納米膠束的粒徑從83.8±13.4nm增加到115.0±12.2nm,納米膠束粒徑有利于藥物遞送到炎癥組織。用巨噬細(xì)胞(RAW264.7)進(jìn)行體外實(shí)驗(yàn),首先,用CCK-8法測(cè)定和評(píng)價(jià)膠束的體外細(xì)胞毒性,結(jié)果表明制備的載體在所用濃度下無(wú)毒。然后,脂多糖(LPS)刺激巨噬細(xì)胞,測(cè)定促炎因子如腫瘤壞死因子(TNF-α)和白細(xì)胞介素-6(IL-6)的細(xì)胞表達(dá),評(píng)價(jià)載藥普通膠束和混合膠束的體外抗炎效果,結(jié)果表明,裝載地塞米松的混合膠束具有比游離地塞米松和載藥普通膠束更好的抗炎效果。最后,巨噬細(xì)胞與游離香豆素或膠束-香豆素共孵育,通過(guò)流式細(xì)胞術(shù)和熒光顯微鏡研究膠束的細(xì)胞攝取,表明混合膠束具有更強(qiáng)的細(xì)胞內(nèi)吞作用。對(duì)小鼠尾根部皮下注射完全弗氏佐劑建立體內(nèi)關(guān)節(jié)炎癥模型,每天給藥,通過(guò)測(cè)量小鼠足跖厚度、小鼠關(guān)節(jié)炎指數(shù)評(píng)分和關(guān)節(jié)的HE染色切片評(píng)價(jià)載藥體系的體內(nèi)抗炎效果,結(jié)果表明載藥膠束能夠明顯的改善小鼠關(guān)節(jié)的紅、腫現(xiàn)象,足跖厚度減少。用近紅外熒光標(biāo)記載體,研究載體在炎癥小鼠體內(nèi)的分布情況,小動(dòng)物活體成像結(jié)果顯示載體能夠靶向于炎癥部位,且熒光強(qiáng)度能夠持續(xù)24h。我們又研究了藥物在體內(nèi)的代謝情況,用PK-solver處理數(shù)據(jù),游離地塞米松原料藥的半衰期為3.49h,載藥普通膠束、載藥混合膠束藥物的半衰期分別為16.52h、19.29h,結(jié)果表明載藥膠束延長(zhǎng)地塞米松在體內(nèi)的循環(huán)時(shí)間。綜上,本研究構(gòu)建的靶向載藥膠束增強(qiáng)了藥物的體外細(xì)胞攝取,提高了藥物的體內(nèi)和體外抗炎效果,具有較高的安全性,有望更加安全、有效地治療類(lèi)風(fēng)濕性關(guān)節(jié)炎。
[Abstract]:Rheumatoid arthritis is a chronic, immune disorder. The incidence of disease is about 1% of the population. The drug delivery system effectively solves the disadvantages of low efficiency and high toxicity of many drugs. Nanoscale drug delivery systems have become a hot research area because of their ability to selectively concentrate in the diseased areas. Natural polysaccharides are nontoxic and biodegradable macromolecular biomaterials, which can be used to prepare nano-sized drug carriers. Polysialic acid (PSAs) is a strong hydrophilic polysaccharide. Cholesterol is an integral part of the cell membrane and is beneficial to the endocytosis of the nano-carrier, which can reduce the uptake of the nanoparticles by the reticuloendothelial system and prolong the circulation time of the drug in the body. In order to improve the delivery of anti-inflammatory drugs, a nanocomposite micelle based on PSA-CC was designed and synthesized to act as a hydrophobic group. The synthesis of mixed micelles was confirmed by NMR-IR IR Size, 味 potential and transmission electron microscope. The critical micelle concentrations of ordinary micelles and targeted mixed micelles based on PSA were 46.2 鹵3.9 渭 g / ml / ml and 32.1 鹵5.2 渭 g / ml / ml, respectively. The classic anti-inflammatory drug dexamethasone is effectively loaded into micelles through hydrophobic interactions. The loading of dexamethasone increased the particle size of targeted mixed micelles from 83.8 鹵13.4 nm to 115.0 鹵12.2 nm. The cytotoxicity of micelle in vitro was determined and evaluated by CCK-8. The results showed that the carrier was not toxic at the concentration used. Then, lipopolysaccharide (LPS) stimulated macrophages, and the expression of proinflammatory factors such as tumor necrosis factor TNF- 偽 (TNF- 偽) and interleukin-6 (IL-6) were measured. The anti-inflammatory effect of common micelles and mixed micelles loaded with Dexamethasone was evaluated in vitro. The results showed that the mixed micelles loaded with dexamethasone had better anti-inflammatory effect than free dexamethasone and drug-loaded micelles. Macrophages were co-incubated with free coumarin or micella-coumarin, and the cellular uptake of micelles was studied by flow cytometry and fluorescence microscopy. The results showed that the mixed micelles had stronger endocytosis. A model of arthritis in vivo was established by subcutaneous injection of complete Freund's adjuvant to the tail root of mice. Mouse arthritis index score and HE staining section of joint were used to evaluate the anti-inflammatory effect of drug delivery system in vivo. The results showed that the micelles could obviously improve the redness and swelling of mouse joints and decrease the thickness of feet and plantar. In vivo imaging of small animals showed that the vector could target the inflammatory site and the fluorescence intensity could last 24 h. We also studied the metabolism of drugs in vivo and processed the data with PK-solver. The half-life of free dexamethasone raw drug was 3.49 h, the half life of drug carrying common micelle and drug mixed micelle was 16.52 h or 19.29 h respectively. The results showed that the drug carrier micelle prolonged the circulation time of dexamethasone in vivo. In this study, the targeted drug carrier micelle enhanced the drug uptake in vitro, improved the anti-inflammatory effect of drugs in vivo and in vitro, and has a higher safety, which is expected to be more safe and effective in the treatment of rheumatoid arthritis.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R943;R96

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