光合細菌益生特性的實驗研究
發(fā)布時間：2018-12-06 12:24
【摘要】：背景: 人類整個胃腸道(gastrointestinal tract,GIT)棲居著大量微生物,這些微生物構成了GIT的生物屏障。但GIT隨時經受各種內源性和外源性的氧化應激,導致胃腸黏膜的氧化損傷。人類已就微生物應對氧化應激開展了少量研究。據(jù)報道一些乳酸桿菌具有抗氧化特性,能減少攝入食物過程中活性氧自由基(reactive oxygen species,ROS)的富集。一些乳酸菌還能降解超氧陰離子自由基和H_2O_2。據(jù)報道類球紅細菌體內外有抗氧化作用,但作用部位不具體,作用機理不確切。尚未見其它種類光合細菌(photosynthetic bacteria,PSB)抗氧化作用的報道,也未見具有抗氧化作用的PSB對氧化損傷胃腸黏膜保護作用的研究報道。 當前PSB作為益生菌株已在國外的保健食品中應用,但尚未列入國內允許用于保健食品的菌種名單內,可見PSB作為益生菌株已有一定應用基礎,但尚缺乏益生特性的系統(tǒng)資料。 目的: 本課題對8株PSB作為益生菌的安全性、功效性、工藝特性等益生特性進行初步研究,并對其進行系統(tǒng)鑒定,同時建立大鼠胃腸黏膜的急性水浸氧化應激模型來探討PSB在宿主體內對氧化損傷胃腸黏膜的保護作用,為開發(fā)PSB保健品、藥品提供益生特性資料和功效開發(fā)方向,以促進PSB在保健品、藥品中的應用。 實驗方法: (一)本課題通過表型特征、培養(yǎng)特征、生理生化特征和菌株的16S rDNA序列分析對8株PSB菌株進行了系統(tǒng)鑒定。(二)觀察8株PSB菌株體外對8種抗生素藥敏紙片的敏感性;對菌株2c(10~(10)CFU/mL)的安全性進行體內實驗,包括急性毒性實驗,檢測血液中內毒素含量,用細菌培養(yǎng)方法觀察攝入菌株2c后是否造成細菌易位,用掃描電鏡和透射電鏡觀察菌株2c對腸黏膜和細胞結構有無不良影響,用試劑盒檢測反映肝腎功能的血液生理生化指標了解菌株2c是否對肝腎功能有不良影響。(三)通過以下方法對PSB菌株的功效性進行了實驗:(1)研究8株PSB菌株體外對酸和膽鹽的耐受、在模擬胃液和小腸液中的存活率、體外對4種腸道常見致病菌的抑制作用和菌株體外對HT-29細胞的黏附作用,以及用Van Niel富集培養(yǎng)基富集培養(yǎng)大鼠攝入PSB期間以及停止攝入PSB后一周之內糞便中的PSB,以預測菌株在宿主體內存活和持續(xù)情況。(2)檢測8
[Abstract]:Background: human gastrointestinal tract (gastrointestinal tract,GIT) inhabit a large number of microorganisms, which constitute the biological barrier of GIT. However, GIT is subjected to various endogenous and exogenous oxidative stress at any time, leading to oxidative injury of gastrointestinal mucosa. Little research has been done on the response of microorganisms to oxidative stress. Some Lactobacilli are reported to have antioxidant properties that reduce the concentration of reactive oxygen free radical (reactive oxygen species,ROS) during food intake. Some lactic acid bacteria can also degrade superoxide anion free radicals and H _ 2O _ 2s _ 2. It is reported that globular red bacteria have antioxidation effect in vivo and in vitro, but the action site is not specific and the mechanism is not exact. There are no reports on the antioxidant effects of other photosynthetic bacteria (photosynthetic bacteria,PSB) or the protective effects of PSB on oxidative injury of gastrointestinal mucosa. Currently, PSB as a probiotic strain has been used in health food abroad, but it has not been included in the list of strains allowed for health food in China. It can be seen that PSB as a probiotic strain has a certain application foundation, but there is still no systematic data on probiotics. Objective: to study the probiotic characteristics of 8 strains of PSB as probiotics, including their safety, efficacy and technological characteristics, and to identify them systematically. At the same time, an acute water-immersion oxidative stress model of rat gastrointestinal mucosa was established to study the protective effect of PSB on oxidative injury of gastrointestinal mucosa in the host, and to provide the information of probiotic characteristics and the development direction of efficacy for the development of PSB health products and drugs. To promote the application of PSB in health products and medicines. Methods: (1) eight strains of PSB were systematically identified by phenotypic, culture, physiological and biochemical characteristics and 16s rDNA sequence analysis. (2) to observe the sensitivity of 8 PSB strains to 8 antibiotic sensitive tablets in vitro; The safety of strain 2c (10 ~ (10) CFU/mL) was tested in vivo, including acute toxicity test, endotoxin content in blood, bacterial culture method was used to observe whether bacterial translocation was caused by strain 2c ingestion. The negative effects of strain 2c on intestinal mucosa and cell structure were observed by scanning electron microscope (SEM) and transmission electron microscope (TEM). The blood physiological and biochemical indexes reflecting liver and kidney function were detected by kit to determine whether strain 2c had adverse effect on liver and kidney function. (3) the efficacy of PSB strains was tested by the following methods: (1) the tolerance of 8 PSB strains to acid and bile salt in vitro and the survival rate in simulated gastric juice and intestinal fluid were studied. Inhibition of four common intestinal pathogens in vitro and adhesion of the strains to HT-29 cells in vitro, and PSB, in feces of rats enriched with Van Niel enrichment medium during uptake of PSB and within one week after cessation of PSB intake To predict the survival and persistence of the strain in the host. (2) Detection 8
【學位授予單位】：第三軍醫(yī)大學
【學位級別】：博士
【學位授予年份】：2005
【分類號】：R37
本文編號：2366021
[Abstract]:Background: human gastrointestinal tract (gastrointestinal tract,GIT) inhabit a large number of microorganisms, which constitute the biological barrier of GIT. However, GIT is subjected to various endogenous and exogenous oxidative stress at any time, leading to oxidative injury of gastrointestinal mucosa. Little research has been done on the response of microorganisms to oxidative stress. Some Lactobacilli are reported to have antioxidant properties that reduce the concentration of reactive oxygen free radical (reactive oxygen species,ROS) during food intake. Some lactic acid bacteria can also degrade superoxide anion free radicals and H _ 2O _ 2s _ 2. It is reported that globular red bacteria have antioxidation effect in vivo and in vitro, but the action site is not specific and the mechanism is not exact. There are no reports on the antioxidant effects of other photosynthetic bacteria (photosynthetic bacteria,PSB) or the protective effects of PSB on oxidative injury of gastrointestinal mucosa. Currently, PSB as a probiotic strain has been used in health food abroad, but it has not been included in the list of strains allowed for health food in China. It can be seen that PSB as a probiotic strain has a certain application foundation, but there is still no systematic data on probiotics. Objective: to study the probiotic characteristics of 8 strains of PSB as probiotics, including their safety, efficacy and technological characteristics, and to identify them systematically. At the same time, an acute water-immersion oxidative stress model of rat gastrointestinal mucosa was established to study the protective effect of PSB on oxidative injury of gastrointestinal mucosa in the host, and to provide the information of probiotic characteristics and the development direction of efficacy for the development of PSB health products and drugs. To promote the application of PSB in health products and medicines. Methods: (1) eight strains of PSB were systematically identified by phenotypic, culture, physiological and biochemical characteristics and 16s rDNA sequence analysis. (2) to observe the sensitivity of 8 PSB strains to 8 antibiotic sensitive tablets in vitro; The safety of strain 2c (10 ~ (10) CFU/mL) was tested in vivo, including acute toxicity test, endotoxin content in blood, bacterial culture method was used to observe whether bacterial translocation was caused by strain 2c ingestion. The negative effects of strain 2c on intestinal mucosa and cell structure were observed by scanning electron microscope (SEM) and transmission electron microscope (TEM). The blood physiological and biochemical indexes reflecting liver and kidney function were detected by kit to determine whether strain 2c had adverse effect on liver and kidney function. (3) the efficacy of PSB strains was tested by the following methods: (1) the tolerance of 8 PSB strains to acid and bile salt in vitro and the survival rate in simulated gastric juice and intestinal fluid were studied. Inhibition of four common intestinal pathogens in vitro and adhesion of the strains to HT-29 cells in vitro, and PSB, in feces of rats enriched with Van Niel enrichment medium during uptake of PSB and within one week after cessation of PSB intake To predict the survival and persistence of the strain in the host. (2) Detection 8
【學位授予單位】：第三軍醫(yī)大學
【學位級別】：博士
【學位授予年份】：2005
【分類號】：R37
【引證文獻】
相關期刊論文 前1條
1 許樂樂;王園園;劉春娥;李永芹;Kevin David Penkava;莊如豪;李澤宇;王學超;;光合細菌功能型胡蘿卜汁飲料的研制[J];食品科技;2013年02期
，本文編號：2366021
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