本文選題：幽門螺桿菌　切入點(diǎn)：MS　出處：《第三軍醫(yī)大學(xué)》2005年博士論文　論文類型：學(xué)位論文

【摘要】：幽門螺桿菌(Helicobacter pylori, Hp)是引起人類慢性胃炎、消化性潰瘍、胃癌等的主要致病菌,現(xiàn)階段在全世界人群中的感染率達(dá)50%以上,目前臨床上常用的三聯(lián)療法不足以根治Hp 感染,且費(fèi)用昂貴、病人依從性差、容易復(fù)發(fā)及產(chǎn)生耐藥菌株。疫苗接種是徹底消除Hp 感染的有效措施之一,根據(jù)共同粘膜免疫系統(tǒng)(common mucosal immunity system, CMIS)理論及文獻(xiàn)報(bào)道,口服途徑是其中最簡單、直接、安全且有效的給藥方式,但必須克服消化道中大量酸性、酶類物質(zhì)對(duì)抗原破壞的問題。 近年來,以微球(microsphere, MS)、微囊、脂質(zhì)體等為代表的緩釋、控釋給藥制劑研究得到迅猛發(fā)展,它不僅能耐受各種復(fù)雜生理環(huán)境,而且還具有靶向給藥的特點(diǎn)。其靶向性決定于制劑本身粒徑大小,有文獻(xiàn)報(bào)道當(dāng)粒徑為lμm 至l0μm 之間的MS經(jīng)口服后,既可以產(chǎn)生全身免疫,也可以在局部粘膜形成有效的免疫應(yīng)答。另外,由于MS 具有顯著的緩釋特性,能夠使藥物在靶器官形成長時(shí)間的有效濃度,因此可以在一定程度上減少疫苗的免疫劑量及免疫次數(shù)。 Hp 疫苗免疫保護(hù)機(jī)理的深入探討對(duì)于Hp 的有效防治及疫苗的研發(fā)會(huì)起到較大的推定作用。目前盡管對(duì)其進(jìn)行了大量的研究,但其確切機(jī)制尚不清楚。本研究以Hp全菌蛋白(HpWCP)為抗原成分,主要從MS 制備工藝及特征分析、免疫效應(yīng)、免疫保護(hù)及保護(hù)機(jī)理等三個(gè)方面進(jìn)行了研究。 (一) HpWCP-CAMS 的制備及其表征分析 在本研究中,采用生物可降解天然高分子物質(zhì)-殼聚糖(chitosan, CTS)、海藻酸鈉(sodium alginate, AGS)為MS 主要制備材料,通過系統(tǒng)考察制備材料水溶液的流動(dòng)性、粘稠度、穩(wěn)定性等結(jié)果確定了AGS 乳液濃度為2%、CTS 濃度為1%、植物油與AGS乳液配比為2:8、CaCl2溶液反滴滴定法等MS 制備工藝參數(shù)及程序。在此基礎(chǔ)上,對(duì)影響MS 粒徑大小的因素進(jìn)行均勻設(shè)計(jì)分析,并以確定的攪拌速率800rpm、藥物濃度2mg/ml、制備溫度25℃、攪拌時(shí)間30min 等參數(shù)制備出平均粒徑為3.33μm 殼聚糖-海藻酸鈉包裹牛血清白蛋白微球(BSA-CAMS),其平均載藥量為31.5%,平均藥物包封率為61.0%。電鏡觀察所得MS 表面光整、大小均一。體外釋藥實(shí)驗(yàn)結(jié)果顯示此MS具有良好的緩釋功能,最長釋藥時(shí)間可達(dá)到20d 以上。在人工胃液中觀察其具有強(qiáng)耐
[Abstract]:Helicobacter pylori (HP) is the main pathogen causing chronic gastritis, peptic ulcer, gastric cancer and so on. At present, the infection rate of Helicobacter pylori is more than 50% in the world population. At present, the triple therapy commonly used in clinical practice is not enough to cure HP infection. And the cost is high, the patient compliance is poor, easy to relapse and to produce drug-resistant strains. Vaccination is one of the effective measures to completely eliminate HP infection, according to the common mucosal immune system, common mucosal immunity system (CMISS) theory and literature report. Oral administration is the most simple, direct, safe and effective way of administration, but it is necessary to overcome the problem that a large number of acidic and enzymatic substances destroy the antigen in the digestive tract. In recent years, the research of microsphere, MSG, microcapsule, liposome and controlled release drug delivery has been developed rapidly, which can not only tolerate various complex physiological environments. Moreover, it has the characteristics of targeted administration. Its targeting depends on the size of the preparation itself. It has been reported that when MS with the particle size of 1 渭 m to 1.0 渭 m is taken orally, it can produce systemic immunity. It can also form an effective immune response in the local mucous membrane. In addition, because MS has a remarkable slow-release property, it can make the effective concentration of the drug in the target organ form for a long time. Therefore, the immunization dose and times of immunization can be reduced to a certain extent. The further study on the mechanism of immunization protection of HP vaccine will play an important role in the effective prevention and treatment of HP and the research and development of the vaccine. Although a great deal of research has been done on it at present, However, the exact mechanism of HpWCPS was not clear. In this study, HpWCPs were used as antigen components, mainly from three aspects: preparation process and characteristic analysis of MS, immune effect, immune protection and protective mechanism. (1) preparation and characterization of HpWCP-CAMS. In this study, the biodegradable natural macromolecules, chitosan chitosan, CTS, alginate sodium alginate, AGSwere used as the main preparation materials of MS. The fluidity and viscosity of aqueous solution of the prepared materials were systematically investigated. The results of stability determined the process parameters and procedures of MS preparation, such as the concentration of AGS emulsion was 2 and CTS concentration was 1, the ratio of vegetable oil to AGS emulsion was 2: 8, CaCl2 solution reverse drop titration, and so on. On the basis of this, the factors affecting the size of MS particle size were designed and analyzed uniformly. The stirring rate was 800rpm, the drug concentration was 2 mg / ml, and the preparation temperature was 25 鈩，

本文編號(hào)：1586500