Identification,Probiotic Characterization and Anti-Helicobac
發(fā)布時(shí)間：2021-12-18 15:19
幽門螺桿菌是引起慢性胃炎,十二指腸潰瘍和胃腫瘤的病原體。它悄無聲息地潛藏在世界50%的人口,是癌癥的主要誘因�？褂拈T螺桿菌菌株的出現(xiàn)是治療感染的又一挑戰(zhàn)。然而即使感染得到治療,復(fù)發(fā)也是一個(gè)普遍的問題。乳酸菌(LAB)的功效已被廣泛研究,用于對(duì)包括幽門螺桿菌感染在內(nèi)的多種疾病的治療產(chǎn)生有益影響。具有潛在益生菌特性的乳酸桿菌是有效用作多種疾病的有益治療劑。益生菌對(duì)病原體的作用涉及不同的機(jī)制,包括免疫調(diào)節(jié),競(jìng)爭性排斥,抗菌素的分泌,微生物群的恢復(fù)。益生菌的作用是特異性的,與每個(gè)菌株特性有關(guān),因此,研究單個(gè)菌株的功效至關(guān)重要。除了慢性炎癥和氧化應(yīng)激反應(yīng),幽門螺桿菌還能引起胃微生物群失衡。益生菌能改善幽門螺桿菌引起的炎癥的功效已被廣泛研究,而益生菌恢復(fù)胃微生物群功效的研究卻很少。這項(xiàng)研究的重點(diǎn)是使用預(yù)處理和研究新鑒定的副干酪乳桿菌ZFM54菌株在幽門螺桿菌感染的小鼠模型中改善幽門螺桿菌介導(dǎo)的炎癥和胃營養(yǎng)不良的潛力。從健康嬰兒的糞便中分離出副干酪乳桿菌ZFM54菌株,進(jìn)行抗幽門螺桿菌潛能測(cè)定,并進(jìn)行全基因組測(cè)序。副干酪乳桿菌ZFM54具有益生菌相關(guān)性,這些基因具有潛在的益生菌特性,表現(xiàn)在無論是在惡...
【文章來源】： 浙江工商大學(xué)浙江省
【文章頁數(shù)】：149 頁
【文章目錄】：
摘要
ABSTRACT
ABBREVIATIONS
CHAPTER 1 INTRODUCTION
1.1 INTRODUCTION
1.2 AIMS AND OBJECTIVES
CHAPTER 2 LITERATURE REVIEW
2.1 HELICOBACTER PYLORI:PATHOGENESIS
2.1.1 Epidemiology
2.1.2 Diagnosis
2.1.2.1 Non-invasive methods
2.1.2.2 Urea Breath Test(UBT)
2.1.2.3 Stool antigen test
2.1.2.4 Serology
2.1.2.5 Invasive tests
2.1.2.5.1 Endoscopy and biopsy
2.1.3 Treatment
2.1.3.1 First line of therapy
2.1.3.2 Second line of therapy
2.1.3.3 Third line of therapy
2.2 DEFENCE MECHANISM OF THE HOST
2.2.1 Microbiota as first line of defence
2.2.1.1 Role of microbiota
2.2.1.2 Dysbiosis
2.2.1.3 Gastric microbiota and H.pylori
2.2.1.4 Gastrointestinal epithelium as second line of defence
2.2.3 Immune system as third line of defence
2.2.3.1 Innate and adaptive immune system(Overview)
2.3 HUMAN STOMACH
2.4 HISTORY OF PROBIOTICS
2.4.1 PROBIOTIC
2.4.2 The genus Lactobacillus
2.4.3 Immune tolerance and immune regulation mechanism of Lactic acid bacteria(General overview)
2.5 MECHANISMS OF ACTION OF PROBIOTICS AGAINST HELICOBACTERPYLORI
2.5.1 Immunomodulation
2.5.2 Co-aggregation and aggregation
2.5.3 Strengthening the mucosal barrier
2.5.4 Competition for adhesion
2.5.5 Secretion of antimicrobials
2.6 EXPERIMENTAL STUDIES OF ANTI-H.PYLORI ACTIVITY OF PROBIOTICS WITH CELL LINESAND ANIMALS
2.7 CLINICAL STUDIES USING PROBIOTIC IN H.PYLORI ERADICATION
2.7.1 Probiotic as an alternative to antibiotics to eradicate H.pylori
2.7.2 Probiotics as an adjunct to antibiotics to eradicate H.pylori
CHAPTER 3 IDENTIFICATION AND PROBIOTIC CHARACTERIZATION OFLACTOBACILLUS PARACASEI ZFM54
3.1 INTRODUCTION
3.2 MATERIALS AND METHODS:
3.2.1 Bacterial growth conditions
3.2.2 Identification of L.paracasei ZFM54
3.2.3 Phylogenetic analysis
3.2.4 Genome sequence
3.2.5 Genome annotation
3.2.6 In vitro studies on probiotics characteristics
3.2.7 Pathogen inhibition by agar well diffusion assay
3.2.8 Tolerance to different concentrations of salt
3.2.9 Tolerance to simulated gastric juice
3.2.10 Tolerance to bile salts
3.2.11 Auto-aggregation assay
3.2.12 Hydrophobicity
3.2.13 Antibiotic susceptibility
3.2.14 Toxicity in mice
3.3 RESULTS
3.3.1 Identification of L.paracaseiZFM54
3.3.2 Genomic and probiotic features
3.3.3 Pathogen inhibition by agar well diffusion assay
3.3.4 Tolerance to different concentration of salt
3.3.5 Tolerance to simulated gastric juice
3.3.6 Tolerance to bile salts
3.3.7 Auto-aggregation assay
3.3.8 Hydrophobicity
3.3.9 Antibiotic susceptibility
3.3.10 Acute Toxicity in mice
3.4 Discussion
Chapter 4 In vitro studies on anti-Helicobacter pylori activity of L. paracasei ZFM54
4.1 INTRODUCTION
4.2 MATERIALS AND METHODS
4.2.1 Bacterial strains and culture conditions
4.2.2 Agar well diffusion assay
4.2.3 Urease inhibition assay
4.2.4 Co-aggregation assay
4.2.5 Anti-oxidant activity
4.3 RESULTS
4.3.1 Agar well diffusion assay
4.3.2 Urease inhibition assay
4.3.3 Co-aggregation
4.3.4 Anti-oxidant activity
4.4 DISCUSSION
CHAPTER 5 IN VIVO STUDIES ON PREVENTIVE AND THERAPEUTIC EFFECT OF L.PARACASEI ZFM54 ON H.PYLORI INFECTED MICE MODEL
5.1 INTRODUCTION
5.2 MATERIALS AND METHODS
5.2.1 In vivo experiments with mice
5.2.2 Bacterial cultures
5.2.3 Animals:
5.2.4 MDA assay
5.2.5 GSH assay
5.2.6 Histopathology of Gastric Tissues
5.2.7 Real-time PCR
5.2.7.1 Reverse transcription(qPCR) Method
5.2.8 16S rRNA gene sequencing
5.2.9 16S rRNA sequencing analysis for microbiota
5.2.10 Statistical Analyses
5.3 RESULTS
5.3.1 Weight of mice during preventive experiment with 54 and LGG
5.3.2 Histopathology of stomach tissues
5.3.3 MDA and GSH level
5.3.4 Effect of pretreatment with strain54 and LGG on inflammatory cytokines
5.3.5 Linear discriminent effect size(LEfSe)analysis of bacterial taxa in preventiongroups
5.3.6 Relative abundance of genus Helicobacter in prevention groups
5.3.7 Weight of mice during therapeutic experiment with strain54,LGG and Triple Therapy
5.3.8 Histopathology of stomach tissues
5.3.9 MDA and GSH level
MDA
GSH
5.3.10 Effect of treatment with strain54,LGG and triple therapy(TT) oninflammatorycytokines
5.3.11 Linear discriminent effect size(LEfSe)analysis of bacterial taxa in therapeuticgroups
5.3.12 Relative abundance of genus Helicobacter in therapeutic groups
5.4 DISCUSSION
Chapter 6 Conclusions
6.1 Overall conclusion
6.2 Summary of conclusions
6.3 Future prospects
REFERENCES
ACKNOWLEDGEMENTS
Publications
【參考文獻(xiàn)】：
期刊論文
[1]Probiotic BIFICO cocktail ameliorates Helicobacter pylori induced gastritis [J]. Hong-Jing Yu,Wei Liu,Zhen Chang,Hui Shen,Li-Juan He,Sha-Sha Wang,Lu Liu,Yuan-Ying Jiang,Guo-Tong Xu,Mao-Mao An,Jun-Dong Zhang. World Journal of Gastroenterology. 2015(21)
[2]Use of probiotics in the fight against Helicobacter pylori [J]. Paolo Ruggiero. World Journal of Gastrointestinal Pathophysiology. 2014(04)
[3]Factors that mediate colonization of the human stomach by Helicobacter pylori [J]. Ciara Dunne,Brendan Dolan,Marguerite Clyne. World Journal of Gastroenterology. 2014(19)
[4]History of Helicobacter pylori,duodenal ulcer,gastric ulcer and gastric cancer [J]. David Y Graham. World Journal of Gastroenterology. 2014(18)
[5]Helicobacter pylori infection: Host immune response, implications on gene expression and microRNAs [J]. Aline Cristina Targa Cadamuro,Ana Flávia Teixeira Rossi,Nathália Maciel Maniezzo,Ana Elizabete Silva. World Journal of Gastroenterology. 2014(06)
[6]A review of Helicobacter pylori diagnosis, treatment, and methods to detect eradication [J]. Elvira Garza-González,Guillermo Ignacio Perez-Perez,Héctor Jesús Maldonado-Garza,Francisco Javier Bosques-Padilla. World Journal of Gastroenterology. 2014(06)
[7]Helicobacter pylori and interleukin-8 in gastric cancer [J]. Ko Eun Lee,Pham Ngoc Khoi,Yong Xia,Jung Sun Park,Young Eun Joo,Kyung Keun Kim,Seok Yong Choi,Young Do Jung. World Journal of Gastroenterology. 2013(45)
[8]Adjuvant probiotics improve the eradication effect of triple therapy for Helicobacter pylori infection [J]. Yi-Qi Du, Tun Su, Jian-Gao Fan, Yu-Xia Lu, Ping Zheng, Xing-Hua Li, Chuan-Yong Guo, Ping Xu, Yan-Fang Gong, Zhao-Shen Li, Department of Gastroenterology, Changhai Hospital, Second Military Medical University, Shanghai 200433, China, Department of Gastroenterology, Xinhua Hospital, Shanghai Jiao Tong University, Shanghai 200240, China, Department of Gastroenterology, Tongji Hospital, Tong Ji University, Shanghai 200092, China, Department of Gastroenterology, Shanghai First People’s Hospital, Shanghai 201620, China, Department of Gastroenterology, Shanghai Eighth People’s Hospital, Shanghai 210035, China , Department of Gastroenterology, Shanghai Tenth People’s Hospital, Shanghai 200072, China, Department of Gastroenterology, Shanghai Songjiang Centeral Hospital, Shanghai 201100, China. World Journal of Gastroenterology. 2012(43)
[9]Lactobacillus plantarum B7 inhibits Helicobacter pylori growth and attenuates gastric inflammation [J]. Chompoonut Sunanliganon,Duangporn Thong-Ngam,Somying Tumwasorn,Naruemon Klaikeaw. World Journal of Gastroenterology. 2012(20)
[10]Lactobacilli inhibit interleukin-8 production induced by Helicobacter pylori lipopolysaccharide-activated Toll-like receptor 4 [J]. Chao Zhou, Feng-Zhen Ma, Xue-Jie Deng, Hong Yuan, Hong-Sheng Ma, Department of Gastroenterology, West China Hospital of Sichuan University, Chengdu 610041, Sichuan Province, China. World Journal of Gastroenterology. 2008(32)
本文編號(hào)：3542663
【文章來源】： 浙江工商大學(xué)浙江省
【文章頁數(shù)】：149 頁
【文章目錄】：
摘要
ABSTRACT
ABBREVIATIONS
CHAPTER 1 INTRODUCTION
1.1 INTRODUCTION
1.2 AIMS AND OBJECTIVES
CHAPTER 2 LITERATURE REVIEW
2.1 HELICOBACTER PYLORI:PATHOGENESIS
2.1.1 Epidemiology
2.1.2 Diagnosis
2.1.2.1 Non-invasive methods
2.1.2.2 Urea Breath Test(UBT)
2.1.2.3 Stool antigen test
2.1.2.4 Serology
2.1.2.5 Invasive tests
2.1.2.5.1 Endoscopy and biopsy
2.1.3 Treatment
2.1.3.1 First line of therapy
2.1.3.2 Second line of therapy
2.1.3.3 Third line of therapy
2.2 DEFENCE MECHANISM OF THE HOST
2.2.1 Microbiota as first line of defence
2.2.1.1 Role of microbiota
2.2.1.2 Dysbiosis
2.2.1.3 Gastric microbiota and H.pylori
2.2.1.4 Gastrointestinal epithelium as second line of defence
2.2.3 Immune system as third line of defence
2.2.3.1 Innate and adaptive immune system(Overview)
2.3 HUMAN STOMACH
2.4 HISTORY OF PROBIOTICS
2.4.1 PROBIOTIC
2.4.2 The genus Lactobacillus
2.4.3 Immune tolerance and immune regulation mechanism of Lactic acid bacteria(General overview)
2.5 MECHANISMS OF ACTION OF PROBIOTICS AGAINST HELICOBACTERPYLORI
2.5.1 Immunomodulation
2.5.2 Co-aggregation and aggregation
2.5.3 Strengthening the mucosal barrier
2.5.4 Competition for adhesion
2.5.5 Secretion of antimicrobials
2.6 EXPERIMENTAL STUDIES OF ANTI-H.PYLORI ACTIVITY OF PROBIOTICS WITH CELL LINESAND ANIMALS
2.7 CLINICAL STUDIES USING PROBIOTIC IN H.PYLORI ERADICATION
2.7.1 Probiotic as an alternative to antibiotics to eradicate H.pylori
2.7.2 Probiotics as an adjunct to antibiotics to eradicate H.pylori
CHAPTER 3 IDENTIFICATION AND PROBIOTIC CHARACTERIZATION OFLACTOBACILLUS PARACASEI ZFM54
3.1 INTRODUCTION
3.2 MATERIALS AND METHODS:
3.2.1 Bacterial growth conditions
3.2.2 Identification of L.paracasei ZFM54
3.2.3 Phylogenetic analysis
3.2.4 Genome sequence
3.2.5 Genome annotation
3.2.6 In vitro studies on probiotics characteristics
3.2.7 Pathogen inhibition by agar well diffusion assay
3.2.8 Tolerance to different concentrations of salt
3.2.9 Tolerance to simulated gastric juice
3.2.10 Tolerance to bile salts
3.2.11 Auto-aggregation assay
3.2.12 Hydrophobicity
3.2.13 Antibiotic susceptibility
3.2.14 Toxicity in mice
3.3 RESULTS
3.3.1 Identification of L.paracaseiZFM54
3.3.2 Genomic and probiotic features
3.3.3 Pathogen inhibition by agar well diffusion assay
3.3.4 Tolerance to different concentration of salt
3.3.5 Tolerance to simulated gastric juice
3.3.6 Tolerance to bile salts
3.3.7 Auto-aggregation assay
3.3.8 Hydrophobicity
3.3.9 Antibiotic susceptibility
3.3.10 Acute Toxicity in mice
3.4 Discussion
Chapter 4 In vitro studies on anti-Helicobacter pylori activity of L. paracasei ZFM54
4.1 INTRODUCTION
4.2 MATERIALS AND METHODS
4.2.1 Bacterial strains and culture conditions
4.2.2 Agar well diffusion assay
4.2.3 Urease inhibition assay
4.2.4 Co-aggregation assay
4.2.5 Anti-oxidant activity
4.3 RESULTS
4.3.1 Agar well diffusion assay
4.3.2 Urease inhibition assay
4.3.3 Co-aggregation
4.3.4 Anti-oxidant activity
4.4 DISCUSSION
CHAPTER 5 IN VIVO STUDIES ON PREVENTIVE AND THERAPEUTIC EFFECT OF L.PARACASEI ZFM54 ON H.PYLORI INFECTED MICE MODEL
5.1 INTRODUCTION
5.2 MATERIALS AND METHODS
5.2.1 In vivo experiments with mice
5.2.2 Bacterial cultures
5.2.3 Animals:
5.2.4 MDA assay
5.2.5 GSH assay
5.2.6 Histopathology of Gastric Tissues
5.2.7 Real-time PCR
5.2.7.1 Reverse transcription(qPCR) Method
5.2.8 16S rRNA gene sequencing
5.2.9 16S rRNA sequencing analysis for microbiota
5.2.10 Statistical Analyses
5.3 RESULTS
5.3.1 Weight of mice during preventive experiment with 54 and LGG
5.3.2 Histopathology of stomach tissues
5.3.3 MDA and GSH level
5.3.4 Effect of pretreatment with strain54 and LGG on inflammatory cytokines
5.3.5 Linear discriminent effect size(LEfSe)analysis of bacterial taxa in preventiongroups
5.3.6 Relative abundance of genus Helicobacter in prevention groups
5.3.7 Weight of mice during therapeutic experiment with strain54,LGG and Triple Therapy
5.3.8 Histopathology of stomach tissues
5.3.9 MDA and GSH level
MDA
GSH
5.3.10 Effect of treatment with strain54,LGG and triple therapy(TT) oninflammatorycytokines
5.3.11 Linear discriminent effect size(LEfSe)analysis of bacterial taxa in therapeuticgroups
5.3.12 Relative abundance of genus Helicobacter in therapeutic groups
5.4 DISCUSSION
Chapter 6 Conclusions
6.1 Overall conclusion
6.2 Summary of conclusions
6.3 Future prospects
REFERENCES
ACKNOWLEDGEMENTS
Publications
【參考文獻(xiàn)】：
期刊論文
[1]Probiotic BIFICO cocktail ameliorates Helicobacter pylori induced gastritis [J]. Hong-Jing Yu,Wei Liu,Zhen Chang,Hui Shen,Li-Juan He,Sha-Sha Wang,Lu Liu,Yuan-Ying Jiang,Guo-Tong Xu,Mao-Mao An,Jun-Dong Zhang. World Journal of Gastroenterology. 2015(21)
[2]Use of probiotics in the fight against Helicobacter pylori [J]. Paolo Ruggiero. World Journal of Gastrointestinal Pathophysiology. 2014(04)
[3]Factors that mediate colonization of the human stomach by Helicobacter pylori [J]. Ciara Dunne,Brendan Dolan,Marguerite Clyne. World Journal of Gastroenterology. 2014(19)
[4]History of Helicobacter pylori,duodenal ulcer,gastric ulcer and gastric cancer [J]. David Y Graham. World Journal of Gastroenterology. 2014(18)
[5]Helicobacter pylori infection: Host immune response, implications on gene expression and microRNAs [J]. Aline Cristina Targa Cadamuro,Ana Flávia Teixeira Rossi,Nathália Maciel Maniezzo,Ana Elizabete Silva. World Journal of Gastroenterology. 2014(06)
[6]A review of Helicobacter pylori diagnosis, treatment, and methods to detect eradication [J]. Elvira Garza-González,Guillermo Ignacio Perez-Perez,Héctor Jesús Maldonado-Garza,Francisco Javier Bosques-Padilla. World Journal of Gastroenterology. 2014(06)
[7]Helicobacter pylori and interleukin-8 in gastric cancer [J]. Ko Eun Lee,Pham Ngoc Khoi,Yong Xia,Jung Sun Park,Young Eun Joo,Kyung Keun Kim,Seok Yong Choi,Young Do Jung. World Journal of Gastroenterology. 2013(45)
[8]Adjuvant probiotics improve the eradication effect of triple therapy for Helicobacter pylori infection [J]. Yi-Qi Du, Tun Su, Jian-Gao Fan, Yu-Xia Lu, Ping Zheng, Xing-Hua Li, Chuan-Yong Guo, Ping Xu, Yan-Fang Gong, Zhao-Shen Li, Department of Gastroenterology, Changhai Hospital, Second Military Medical University, Shanghai 200433, China, Department of Gastroenterology, Xinhua Hospital, Shanghai Jiao Tong University, Shanghai 200240, China, Department of Gastroenterology, Tongji Hospital, Tong Ji University, Shanghai 200092, China, Department of Gastroenterology, Shanghai First People’s Hospital, Shanghai 201620, China, Department of Gastroenterology, Shanghai Eighth People’s Hospital, Shanghai 210035, China , Department of Gastroenterology, Shanghai Tenth People’s Hospital, Shanghai 200072, China, Department of Gastroenterology, Shanghai Songjiang Centeral Hospital, Shanghai 201100, China. World Journal of Gastroenterology. 2012(43)
[9]Lactobacillus plantarum B7 inhibits Helicobacter pylori growth and attenuates gastric inflammation [J]. Chompoonut Sunanliganon,Duangporn Thong-Ngam,Somying Tumwasorn,Naruemon Klaikeaw. World Journal of Gastroenterology. 2012(20)
[10]Lactobacilli inhibit interleukin-8 production induced by Helicobacter pylori lipopolysaccharide-activated Toll-like receptor 4 [J]. Chao Zhou, Feng-Zhen Ma, Xue-Jie Deng, Hong Yuan, Hong-Sheng Ma, Department of Gastroenterology, West China Hospital of Sichuan University, Chengdu 610041, Sichuan Province, China. World Journal of Gastroenterology. 2008(32)
本文編號(hào)：3542663
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