胆汁纳米细菌感染致兔胆囊黑色素结石的实验研究
发布时间：2018-12-17 21:56
【摘要】：研究背景 胆结石病是消化系统常见疾病,对人民健康危害极大。胆结石分为三大类,即胆固醇结石、棕色胆色素结石和黑色胆色素结石。一般认为:胆固醇结石的发生与胆固醇代谢异常有关,并受胆汁中促成核因子和抑成核因子的共同影响;棕色胆色素结石由细菌感染引起,细菌产生的外源性β-葡萄糖醛酸苷酶水解双葡萄糖醛酸结合胆红素生成游离胆红素和葡萄糖醛酸,前者与钙离子结合形成胆红素钙沉淀,同时,细菌还产生磷脂酶,降解卵磷脂生成棕榈酸,与Ca~(2+)形成棕榈酸钙沉淀,导致棕色胆石形成;黑色胆结石,呈黑色不规则外形,质地坚硬,主要由胆红素盐和粘蛋白构成,病因不明,多发生于溶血性疾病和肝硬化患者的胆囊内。 芬兰科学家Kaiander等进行哺乳动物细胞培养时发现细胞内存在一种超微生物,能通过100nm的滤菌器,根据其体型微小、栖息在血液中的特点,将其命名为Nanobacterium sanguineum,简称Nanobacteria,中文译名纳米细菌。纳米细菌是一种非常古老的超微细菌,不能利用葡萄糖,主要靠摄取氨基酸来提供能量,新陈代谢率极其缓慢,约为普通细菌的1/10,000,当生存环境不利时,能利用环境中的钙、磷合成羟基碳酸磷灰石将自身包裹,形成一层厚厚的生物被膜,使其渡过困难时期。纳米细菌主要寄生于哺乳动物的血液中,由于能透过0.1um滤菌膜,因此在80%的商用牛血清中存在纳米细菌污染,是导致培养细胞不明原因死亡的重要因素。普通的细菌学方法很难发现纳米细菌的存在,相差显微镜和电子显微镜可以清楚地对其进行观测。大量纳米细菌通过受体介导的胞饮作用侵入培养细胞后,被不完全消化,释放出内毒素,导致受感染细胞凋亡。研究表明纳米细菌与肾结石、胆结石、前列腺炎、多囊肾、多囊肝、动脉粥样硬化等疾病的发生有关。研究证实,纳米细菌感染可以导致肾结石的发生:另外,胆囊结石患者血液中纳米细菌的阳性率是非结石患者的4倍,而胆囊结石患者胆汁纳米细菌感染阳性率则高达61.3%,提示胆汁纳米细菌感染可能是诱发胆囊结石的重要原因,因此我们设计本实验,观察胆汁纳米细菌感染对实验兔胆囊结石的诱发作用。我们将实验分两部分进行:第一部分,从胆囊结石患者胆汁中培养纳米细菌,对其微生物学特性进行观察,按文献方法,对其存在的真实性进行鉴定:第二部分,将由人胆汁中分离、培养的纳米细菌注入实验兔的胆囊中,两周后观察动物成石情况,并对动物的胆囊、胆汁、胆石进行分析。
[Abstract]:Background gallstone disease is a common disease of digestive system, which is harmful to people's health. Gallstones are divided into three categories: cholesterol stone, brown pigment stone and black pigment stone. It is generally believed that the occurrence of cholesterol stones is related to the abnormal metabolism of cholesterol, and is affected by both nuclear factors and inhibitory factors in bile. Brown gallstone was caused by bacterial infection. The bacteria produced exogenous 尾 -glucuronidase to hydrolyze diglucuronic acid-conjugated bilirubin to form free bilirubin and glucuronic acid, the former combined with calcium to form calcium bilirubin precipitate. At the same time, bacteria also produced phospholipase, degradation of lecithin to palmitic acid, and Ca~ (2) to form calcium palmitate precipitation, resulting in brown gallstone formation; Black gallstones, with irregular black appearance and hard texture, are mainly composed of bilirubin salt and mucin. The etiology is unknown and occurs in the gallbladder of patients with hemolytic diseases and cirrhosis. In mammalian cell culture, Finnish scientist Kaiander et al. found that there is a kind of ultramicro organism in the cell, which can be called Nanobacteria, by the filter of 100nm, according to its tiny size and perched in blood. Nanometer bacteria. The nanobacteria are very old, ultramicro bacteria that don't use glucose, they're mainly powered by amino acids, and their metabolism is extremely slow, about a tenth of a percent of the normal bacteria, when the environment is bad. It can make use of calcium and phosphorus to synthesize hydroxycarbonate apatite and form a thick biofilm to make it through difficult period. Nanobacteria are mainly parasitic in mammalian blood. Because they can pass through the 0.1um filter membrane, there is contamination of nanobacteria in 80% commercial bovine serum, which is an important factor to cause the unexplained death of cultured cells. It is difficult to find nanobacteria by ordinary bacteriological methods and can be clearly observed by phase contrast microscope and electron microscope. A large number of nanobacteria invaded the cultured cells through receptor-mediated cytosolic action and were incompletely digested and released endotoxin which led to the apoptosis of infected cells. Studies have shown that nano-bacteria are associated with kidney stones, gallstones, prostatitis, polycystic kidney, polycystic liver, atherosclerosis and other diseases. Studies have shown that nanobacterial infections can lead to kidney stones: in addition, the positive rate of nanobacteria in the blood of patients with cholecystolithiasis is four times as high as that of non-stone patients. The positive rate of bile nanobacterial infection in gallstone patients is as high as 61.3, which suggests that bile nanobacterial infection may be an important cause of inducing gallstone, so we designed this experiment. To observe the inducing effect of bile nanobacterial infection on gallstone in rabbits. We divided the experiment into two parts: in the first part, nanobacteria were cultured from bile of cholecystolithiasis patients, their microbiological characteristics were observed, and the authenticity of their existence was identified according to the method of literature. The nanobacteria isolated from human bile were injected into the gallbladder of experimental rabbits. After two weeks, the animal stone formation was observed and the gallbladder, bile and cholelithiasis of animals were analyzed.
【学位授予单位】：汕头大学
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R363
[Abstract]:Background gallstone disease is a common disease of digestive system, which is harmful to people's health. Gallstones are divided into three categories: cholesterol stone, brown pigment stone and black pigment stone. It is generally believed that the occurrence of cholesterol stones is related to the abnormal metabolism of cholesterol, and is affected by both nuclear factors and inhibitory factors in bile. Brown gallstone was caused by bacterial infection. The bacteria produced exogenous 尾 -glucuronidase to hydrolyze diglucuronic acid-conjugated bilirubin to form free bilirubin and glucuronic acid, the former combined with calcium to form calcium bilirubin precipitate. At the same time, bacteria also produced phospholipase, degradation of lecithin to palmitic acid, and Ca~ (2) to form calcium palmitate precipitation, resulting in brown gallstone formation; Black gallstones, with irregular black appearance and hard texture, are mainly composed of bilirubin salt and mucin. The etiology is unknown and occurs in the gallbladder of patients with hemolytic diseases and cirrhosis. In mammalian cell culture, Finnish scientist Kaiander et al. found that there is a kind of ultramicro organism in the cell, which can be called Nanobacteria, by the filter of 100nm, according to its tiny size and perched in blood. Nanometer bacteria. The nanobacteria are very old, ultramicro bacteria that don't use glucose, they're mainly powered by amino acids, and their metabolism is extremely slow, about a tenth of a percent of the normal bacteria, when the environment is bad. It can make use of calcium and phosphorus to synthesize hydroxycarbonate apatite and form a thick biofilm to make it through difficult period. Nanobacteria are mainly parasitic in mammalian blood. Because they can pass through the 0.1um filter membrane, there is contamination of nanobacteria in 80% commercial bovine serum, which is an important factor to cause the unexplained death of cultured cells. It is difficult to find nanobacteria by ordinary bacteriological methods and can be clearly observed by phase contrast microscope and electron microscope. A large number of nanobacteria invaded the cultured cells through receptor-mediated cytosolic action and were incompletely digested and released endotoxin which led to the apoptosis of infected cells. Studies have shown that nano-bacteria are associated with kidney stones, gallstones, prostatitis, polycystic kidney, polycystic liver, atherosclerosis and other diseases. Studies have shown that nanobacterial infections can lead to kidney stones: in addition, the positive rate of nanobacteria in the blood of patients with cholecystolithiasis is four times as high as that of non-stone patients. The positive rate of bile nanobacterial infection in gallstone patients is as high as 61.3, which suggests that bile nanobacterial infection may be an important cause of inducing gallstone, so we designed this experiment. To observe the inducing effect of bile nanobacterial infection on gallstone in rabbits. We divided the experiment into two parts: in the first part, nanobacteria were cultured from bile of cholecystolithiasis patients, their microbiological characteristics were observed, and the authenticity of their existence was identified according to the method of literature. The nanobacteria isolated from human bile were injected into the gallbladder of experimental rabbits. After two weeks, the animal stone formation was observed and the gallbladder, bile and cholelithiasis of animals were analyzed.
【学位授予单位】：汕头大学
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R363
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