【摘要】：目的 幽門螺桿菌(Helicobacter pylori, Hp)作為慢性胃炎、消化性潰瘍甚至胃癌的主要致病因素,至今仍沒有理想的根治方法。微生態(tài)療法從一個(gè)嶄新的角度擬解決傳統(tǒng)療法存在的各種問題,而且微生態(tài)調(diào)節(jié)劑的作用不僅僅在于抗感染,還能調(diào)節(jié)機(jī)體的免疫機(jī)能,改善胃腸道微生態(tài)環(huán)境,促進(jìn)健康。本課題在國內(nèi)外疫苗發(fā)展和乳酸桿菌防治幽門螺桿菌(Hp)成功經(jīng)驗(yàn)的基礎(chǔ)上,直接從胃腸道標(biāo)本中篩選出具有良好抑制或殺滅幽門螺桿菌且具有高度胃粘膜粘附能力的乳酸桿菌,并進(jìn)行鑒定。為開發(fā)優(yōu)良的抑制幽門螺桿菌的微生態(tài)制劑及相關(guān)的保健食品,提供良好的乳酸桿菌菌種;為研究和發(fā)展Hp基因工程生物活菌疫苗提供良好的載體釋放系統(tǒng)。同時(shí)初步探討乳酸桿菌抑制Hp粘附胃粘膜的機(jī)理。 [方法] 1、乳酸桿菌的分離與初步鑒定:采集臨床胃病患者胃粘膜組織的活檢標(biāo)本和胃液標(biāo)本,以及正常人糞便標(biāo)本,采用選擇性培養(yǎng)基分離可疑乳酸桿菌并進(jìn)行初步鑒定。 2、篩選具有良好抑制或殺滅幽門螺桿菌并具有較強(qiáng)胃粘膜粘附能力的乳酸桿菌: 2.1 采用打孔法篩選對Hp抑菌力強(qiáng)的乳酸桿菌菌株。 2.2 對篩選出的乳酸桿菌進(jìn)行微量生化反應(yīng),進(jìn)一步驗(yàn)證。 2.3 選用胃粘膜上皮細(xì)胞SGC7901,進(jìn)行乳酸桿菌粘附能力實(shí)驗(yàn)。 2.4 乳酸桿菌的耐酸耐膽鹽能力檢測:對最后篩選出的乳酸桿菌進(jìn)行耐酸耐膽鹽能力試驗(yàn)。 3、乳酸桿菌抑制Hp粘附胃粘膜機(jī)理的初步探討:通過幽門螺桿菌標(biāo)準(zhǔn)株SS1粘附胃粘膜上皮細(xì)胞SGC7901的細(xì)胞模型,利用間接熒光抗體試驗(yàn),建立乳酸桿菌對Hp粘附SGC7901細(xì)胞的抑制試驗(yàn),并檢測乳酸桿菌活菌和滅活菌株
[Abstract]:Objective Helicobacter pylori (Helicobacter pylori, Hp) is the main cause of chronic gastritis, peptic ulcer and even gastric cancer. From a new point of view, microecological therapy is intended to solve the problems existing in traditional therapies, and the role of microecological regulators is not only in anti-infection, but also in regulating the immune function of the body, improving the gastrointestinal microecological environment and promoting health. Based on the domestic and foreign vaccine development and the successful experience of Lactobacillus in the prevention and treatment of Helicobacter pylori (Hp), The Lactobacillus with good inhibition or killing ability of Helicobacter pylori and high adhesion ability of gastric mucosa were screened directly from gastrointestinal specimens and identified. It can provide a good strain of Lactobacillus for the development of a good microecological preparation for inhibiting Helicobacter pylori and related health food, and a good vector release system for the research and development of Hp genetically engineered live bacteria vaccine. At the same time, the mechanism of inhibition of Hp adhesion to gastric mucosa by Lactobacillus was discussed. [methods] 1. Isolation and preliminary identification of Lactobacillus: biopsy and gastric juice samples from gastric mucosa of patients with clinical gastropathy and fecal specimens from normal people were collected. The suspected Lactobacillus was isolated on selective medium and identified preliminarily. 2. Screening of Lactobacillus with good inhibition or killing ability of Helicobacter pylori and strong adherence to gastric mucosa. 2.1 Lactobacillus strains with strong bacteriostasis to Hp were screened by perforation method. 2.2 trace biochemical reaction of the selected Lactobacillus was carried out and further verified. 2.3 the adhesion ability of Lactobacillus in gastric mucosal epithelial cells (SGC7901,) was tested. 2.4 determination of acid-tolerant bile salt tolerance of Lactobacillus: acid and bile salt tolerance test was performed on the last selected Lactobacillus. 3. The mechanism of inhibition of Hp adhesion to gastric mucosa by Lactobacillus: through the cell model of SGC7901 adhesion of Helicobacter pylori standard strain SS1 to gastric mucosal epithelial cells, indirect fluorescent antibody test was used. The inhibition test of Lactobacillus on Hp adhesion to SGC7901 cells was established, and the live and inactivated Lactobacillus strains were detected.
【學(xué)位授予單位】：四川大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2005
【分類號】：R378

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