膽汁納米細菌感染致兔膽囊黑色素結石的實驗研究
發(fā)布時間：2018-12-17 21:56
【摘要】：研究背景 膽結石病是消化系統(tǒng)常見疾病,對人民健康危害極大。膽結石分為三大類,即膽固醇結石、棕色膽色素結石和黑色膽色素結石。一般認為:膽固醇結石的發(fā)生與膽固醇代謝異常有關,并受膽汁中促成核因子和抑成核因子的共同影響;棕色膽色素結石由細菌感染引起,細菌產生的外源性β-葡萄糖醛酸苷酶水解雙葡萄糖醛酸結合膽紅素生成游離膽紅素和葡萄糖醛酸,前者與鈣離子結合形成膽紅素鈣沉淀,同時,細菌還產生磷脂酶,降解卵磷脂生成棕櫚酸,與Ca~(2+)形成棕櫚酸鈣沉淀,導致棕色膽石形成;黑色膽結石,呈黑色不規(guī)則外形,質地堅硬,主要由膽紅素鹽和粘蛋白構成,病因不明,多發(fā)生于溶血性疾病和肝硬化患者的膽囊內。 芬蘭科學家Kaiander等進行哺乳動物細胞培養(yǎng)時發(fā)現(xiàn)細胞內存在一種超微生物,能通過100nm的濾菌器,根據其體型微小、棲息在血液中的特點,將其命名為Nanobacterium sanguineum,簡稱Nanobacteria,中文譯名納米細菌。納米細菌是一種非常古老的超微細菌,不能利用葡萄糖,主要靠攝取氨基酸來提供能量,新陳代謝率極其緩慢,約為普通細菌的1/10,000,當生存環(huán)境不利時,能利用環(huán)境中的鈣、磷合成羥基碳酸磷灰石將自身包裹,形成一層厚厚的生物被膜,使其渡過困難時期。納米細菌主要寄生于哺乳動物的血液中,由于能透過0.1um濾菌膜,因此在80%的商用牛血清中存在納米細菌污染,是導致培養(yǎng)細胞不明原因死亡的重要因素。普通的細菌學方法很難發(fā)現(xiàn)納米細菌的存在,相差顯微鏡和電子顯微鏡可以清楚地對其進行觀測。大量納米細菌通過受體介導的胞飲作用侵入培養(yǎng)細胞后,被不完全消化,釋放出內毒素,導致受感染細胞凋亡。研究表明納米細菌與腎結石、膽結石、前列腺炎、多囊腎、多囊肝、動脈粥樣硬化等疾病的發(fā)生有關。研究證實,納米細菌感染可以導致腎結石的發(fā)生:另外,膽囊結石患者血液中納米細菌的陽性率是非結石患者的4倍,而膽囊結石患者膽汁納米細菌感染陽性率則高達61.3%,提示膽汁納米細菌感染可能是誘發(fā)膽囊結石的重要原因,因此我們設計本實驗,觀察膽汁納米細菌感染對實驗兔膽囊結石的誘發(fā)作用。我們將實驗分兩部分進行:第一部分,從膽囊結石患者膽汁中培養(yǎng)納米細菌,對其微生物學特性進行觀察,按文獻方法,對其存在的真實性進行鑒定:第二部分,將由人膽汁中分離、培養(yǎng)的納米細菌注入實驗兔的膽囊中,兩周后觀察動物成石情況,并對動物的膽囊、膽汁、膽石進行分析。
[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.
【學位授予單位】：汕頭大學
【學位級別】：博士
【學位授予年份】：2005
【分類號】：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.
【學位授予單位】：汕頭大學
【學位級別】：博士
【學位授予年份】：2005
【分類號】：R363
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