【摘要】：人體膽汁中主要包含膽鹽、卵磷脂和膽固醇這三種物質(zhì)，其中膽固醇在膽汁中的溶解、成核與生長主要受其他兩種物質(zhì)含量變化的影響。當(dāng)前對膽固醇結(jié)石的研究主要集中于在體外進(jìn)行模擬，即采用類似于人體膽汁的模擬膽汁，通過實驗獲得膽固醇結(jié)石的產(chǎn)生機制。 本文的研究主要是從膽固醇在模擬膽汁中的溶解、成核發(fā)生機制和成核時間、以及膽汁中可能影響膽固醇成核的因素這幾個方面著手。 首先，通過制備四元體系（膽鹽-膽固醇-卵磷脂-水）的模擬膽汁，測定膽固醇在該體系中的溶解度（膽汁能夠溶解膽固醇的量），獲得了膽汁中卵磷脂、膽鹽的含量變化以及模擬膽汁總脂度（膽汁中膽鹽、膽固醇與卵磷脂總量之和）對膽固醇溶解度的影響情況。其中，膽汁中各組分的含量通過高效液相色譜法（HPLC）測定。結(jié)果表明，當(dāng)膽鹽含量越高，膽固醇在模擬膽汁中的溶解度越大；膽鹽/卵磷脂摩爾比越大，膽固醇在模擬膽汁中溶解度也變大，摩爾比在0.5-1.77之間時，溶解度達(dá)到最大，當(dāng)摩爾比繼續(xù)增大時，膽固醇的溶解度則逐漸減�。淮送�，隨著膽汁總脂度的增大，膽汁對膽固醇的溶解能力逐漸增強。最后，繪制了膽固醇在三元相圖中的飽和溶解度曲線。 其次，通過顯微鏡法、光散射法考察了不同組成的膽汁中各分散相（包括膠束和囊泡）的粒度分布情況。實驗發(fā)現(xiàn)，對于含有復(fù)雜體系的膽汁，其組分含量的不同，標(biāo)志著膽汁中存在著不同的相態(tài)。其中，模擬膽汁中囊泡的平均粒徑主要在455.5nm左右，膠束的平均粒徑主要在5.7nm左右。此外，通過實驗觀察模擬膽汁中膽固醇晶核的形成過程發(fā)現(xiàn)，膽汁中囊泡出現(xiàn)到囊泡發(fā)生聚集，膽固醇嵌入到囊泡中成核，最終形成液態(tài)晶體或固態(tài)晶體。 最后，由于人體膽汁中的成分比較復(fù)雜，不僅含有以上三種主要物質(zhì)，還包括鈣鹽、蛋白質(zhì)、膽紅素、細(xì)菌以及其他常量元素和微量元素等，所以本文還研究了鈣鹽和蛋白質(zhì)對膽固醇成核的影響情況。 對于鈣鹽的影響，本文以四元體系所構(gòu)成的模擬膽汁作為參照膽汁，添加Ca2+（氯化鈣）后得到的五元體系模擬膽汁（膽鹽-膽固醇-卵磷脂-水-氯化鈣），其成核時間相對變短，膽汁中膽固醇的溶解度減小。在不同總脂度下，Ca2+的濃度越大，溶解度愈小、成核時間愈短，而且，隨著總脂度的增大，成核也越來越快。另外，含鈣的模擬膽汁相圖中其單相區(qū)域變小。因而，基于膽固醇成核時間和溶解度的測定結(jié)果，我們認(rèn)為，Ca2+在膽固醇成核的過程中以促成核因子存在。通過粒度分布分析得到，含鈣的模擬膽汁的囊泡平均粒徑較大。 對于蛋白質(zhì)的影響，本文選擇了牛血清白蛋白（BSA）作為膽汁蛋白質(zhì)，將BSA替換鈣鹽，添加到四元體系的模擬膽汁中，構(gòu)成另一五元體系。溶解度和成核時間測定結(jié)果表明，，BSA通過靜電和疏水作用與卵磷脂結(jié)合，從而影響了卵磷脂形成的囊泡的穩(wěn)定性，膽固醇的溶解度因而隨著BSA濃度的升高而減小。但添加BSA后，膽汁中膽固醇的成核時間并沒有明顯的變化，即BSA的存在，僅對膽固醇的溶解度存在一定的影響，但對膽固醇的成核過程影響很小。
[Abstract]:The bile of human body mainly contains three substances of bile salt, lecithin and cholesterol, wherein the dissolution, nucleation and growth of cholesterol in the bile are mainly influenced by the change of other two substances. The present study on the cholesterol gallstone is mainly focused on the in vitro simulation, that is to use the simulated bile similar to the human bile, and the generation mechanism of the cholesterol stone is obtained through the experiment. The study of this paper is mainly about the dissolution of cholesterol in the simulated bile, the mechanism of nucleation and the time of nucleation, and the factors that may affect the nucleation of cholesterol in the bile. In the first place, the solubility of cholesterol in the system (the amount of bile capable of dissolving cholesterol) was determined by the preparation of the simulated bile of the quaternary system (bile salt-cholesterol-lecithin-water), and the bladder was obtained. The changes in the content of lecithin and bile salt in the juice and the total lipid of the bile (the sum of the total amount of bile salt, cholesterol and lecithin) in the bile to the solubility of cholesterol The content of each component in the bile is determined by high performance liquid chromatography (HP The results showed that the higher the bile salt content, the higher the solubility of cholesterol in the simulated bile, the higher the bile salt/ lecithin molar ratio, the higher the solubility of the cholesterol in the simulated bile and the molar ratio of 0. 5-1.77, when the molar ratio When continued to increase, the solubility of cholesterol is gradually reduced; in addition, with the increase of the total lipid of the bile, the dissolution of the bile to the cholesterol The force is gradually enhanced. Finally, the saturation of cholesterol in the ternary phase diagram is plotted. Solubility curve. Secondly, the disperse phase (including micelles and vesicles) in the bile of different composition was investigated by the method of microscope and light scattering. The results showed that in the bile with complex system, the content of the components of the bile is different, and the contents of the bile are marked in the bile. The average particle size of the vesicles in the simulated bile is about 455. 5nm, and the average particle size of the micelles is mainly about 455. 5nm. In addition, the formation of the cholesterol crystal nucleus in the simulated bile was observed by the experiment. The formation of the cholesterol crystal nucleus in the bile was observed. The formation of the vesicles in the bile and the formation of the cholesterol in the vesicles and the formation of the liquid were found. finally, because the composition of human bile is more complex, not only contains the above three main substances, but also contains calcium salt, protein, bilirubin, bacteria and other constant elements and trace elements, so the calcium salt and the protein are also studied in this paper. The effects of calcium salt on cholesterol nucleation were studied. The five-membered system of bile (bile salt-cholesterol-lecithin-water-water-water-water-water-water-water-water-water-water-water-water-water-water-water-water-water-water-water-water-water) was prepared by using the simulated bile composed of the four-element system as the reference bile and the addition of Ca2 + (calcium chloride). Calcium chloride), and the nucleation time of the calcium chloride is relatively variable. The higher the concentration of Ca2 +, the smaller the solubility, the shorter the nucleation time, and the shorter the nucleation time. The increase of the total lipid and the faster the nucleation. In addition, the calcium-containing The single-phase region of the simulated bile phase diagram is small. Thus, based on the determination of the time and solubility of cholesterol nucleation, we believe that Ca2 + is in cholesterol in the process of nucleation to contribute to the presence of a nuclear factor, obtained by particle size distribution analysis, In this paper, bovine serum albumin (BSA) was selected as the bile protein, and the BSA was replaced with calcium salt and added to the quaternion. In the simulated bile of the system, another five-membered system was constructed. The results of the determination of the solubility and the nucleation time showed that the BSA was combined with the lecithin by static and hydrophobic interactions, thus affecting the stability of the vesicles formed by the lecithin and the dissolution of cholesterol. The degree of solution is thus reduced with the increase of the concentration of BSA. However, after the addition of BSA, there is no significant change in the nucleation time of the cholesterol in the bile, i.e., the presence of BSA, and there is only a certain degree of solubility in the solubility of the cholesterol.
【學(xué)位授予單位】：江南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R575.62

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 陳強譜;郭瑋;;膽汁中的蛋白質(zhì)[J];濱州醫(yī)學(xué)院學(xué)報;1991年01期

2 方馳華;常旭;魯朝敏;楊劍;項楠;鮑蘇蘇;彭豐平;潘家輝;;肝內(nèi)外膽管結(jié)石64排CT數(shù)據(jù)三維重建及其臨床意義[J];南方醫(yī)科大學(xué)學(xué)報;2008年03期

3 莊杉林;肝膽結(jié)石中醫(yī)辨證分型與B超所見關(guān)系臨床研究[J];福建中醫(yī)藥;2000年04期

4 曾繁清,海匯,金利凡;人體膽結(jié)石紅外光譜的比較分析[J];光譜學(xué)與光譜分析;2001年03期

5 舒曄;膽汁中促成核及抗成核因子的研究進(jìn)展[J];國外醫(yī)學(xué)(消化系疾病分冊);1996年01期

6 潘留勇;;膽結(jié)石治療方法的研究進(jìn)展[J];中國醫(yī)藥科學(xué);2011年17期

7 孫穎,楊展瀾,申國榮,周勇,吳瑾光,徐光憲,周孝思;膽結(jié)石組成和形成機理的研究進(jìn)展[J];中國科學(xué)(B輯 化學(xué));2001年05期

8 歐陽健明;張惠敏;;膽結(jié)石研究中的化學(xué)基礎(chǔ)及其研究進(jìn)展[J];臨床肝膽病雜志;2006年02期

9 王炳生;膽結(jié)石碎石和溶石治療現(xiàn)狀[J];臨床外科雜志;2001年03期

10 王琛,張英華,李嚴(yán),安學(xué)勤;模擬人體膽汁相平衡研究的初步探索[J];南京師大學(xué)報(自然科學(xué)版);2003年02期

相關(guān)博士學(xué)位論文 前1條

1 孔靜;肝細(xì)胞膽小管側(cè)膜轉(zhuǎn)運蛋白表達(dá)改變與膽色素結(jié)石形成的相關(guān)性實驗研究[D];中國醫(yī)科大學(xué);2009年

本文編號：2337309