体外抑制幽门螺杆菌的人源乳酸杆菌的选育

发布时间：2018-12-14 06:36

【摘要】：目的幽门螺杆菌(Helicobacter pylori, Hp)作为慢性胃炎、消化性溃疡甚至胃癌的主要致病因素,至今仍没有理想的根治方法。微生态疗法从一个崭新的角度拟解决传统疗法存在的各种问题,而且微生态调节剂的作用不仅仅在于抗感染,还能调节机体的免疫机能,改善胃肠道微生态环境,促进健康。本课题在国内外疫苗发展和乳酸杆菌防治幽门螺杆菌(Hp)成功经验的基础上,直接从胃肠道标本中筛选出具有良好抑制或杀灭幽门螺杆菌且具有高度胃粘膜粘附能力的乳酸杆菌,并进行鉴定。为开发优良的抑制幽门螺杆菌的微生态制剂及相关的保健食品,提供良好的乳酸杆菌菌种;为研究和发展Hp基因工程生物活菌疫苗提供良好的载体释放系统。同时初步探讨乳酸杆菌抑制Hp粘附胃粘膜的机理。 [方法] 1、乳酸杆菌的分离与初步鉴定:采集临床胃病患者胃粘膜组织的活检标本和胃液标本,以及正常人粪便标本,采用选择性培养基分离可疑乳酸杆菌并进行初步鉴定。 2、筛选具有良好抑制或杀灭幽门螺杆菌并具有较强胃粘膜粘附能力的乳酸杆菌: 2.1 采用打孔法筛选对Hp抑菌力强的乳酸杆菌菌株。 2.2 对筛选出的乳酸杆菌进行微量生化反应,进一步验证。 2.3 选用胃粘膜上皮细胞SGC7901,进行乳酸杆菌粘附能力实验。 2.4 乳酸杆菌的耐酸耐胆盐能力检测:对最后筛选出的乳酸杆菌进行耐酸耐胆盐能力试验。 3、乳酸杆菌抑制Hp粘附胃粘膜机理的初步探讨:通过幽门螺杆菌标准株SS1粘附胃粘膜上皮细胞SGC7901的细胞模型,利用间接荧光抗体试验,建立乳酸杆菌对Hp粘附SGC7901细胞的抑制试验,并检测乳酸杆菌活菌和灭活菌株
[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.
【学位授予单位】：四川大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R378

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