本文選題：糞便菌群　切入點(diǎn)：牛奶蛋白過敏　出處：《重慶醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:將牛奶蛋白過敏嬰兒和正常嬰兒的糞便菌群分別移植給無菌小鼠后,構(gòu)建菌群人源化小鼠模型,觀察移植小鼠對(duì)牛奶蛋白的反應(yīng),探究糞便菌群在牛奶蛋白過敏發(fā)生中的作用。方法:收集重慶醫(yī)科大學(xué)附屬兒童醫(yī)院消化�？婆R床診斷為牛奶蛋白過敏(cow’milk protein allergy,CMPA)且以胃腸道表現(xiàn)為主的嬰兒糞便(I-CMPA組),同時(shí)收集該院兒�？企w檢的、無胃腸道癥狀且無過敏性疾病的正常嬰兒糞便(I-NC組),分別移植給無菌小鼠后為過敏移植小鼠組(M-CMPA組)和正常移植小鼠組(M-NC組),兩組移植小鼠均用嬰兒配方奶粉喂養(yǎng)。14天后,用Illumina測(cè)序技術(shù)檢測(cè)糞便菌群的定植規(guī)律和變化情況、觀察移植小鼠的臨床表現(xiàn)、HE染色觀察空腸組織的病理改變、流式細(xì)胞術(shù)檢測(cè)脾臟細(xì)胞中CD4+CD25+Foxp3+調(diào)節(jié)性T細(xì)胞比例變化。結(jié)果:(1)Illumina測(cè)序結(jié)果分析顯示,供體糞菌間的優(yōu)勢(shì)菌不同,且I-CMPA組菌屬豐富度和多樣性均較I-NC組低。糞菌移植14天后,I-CMPA組中的重要組分能在無菌小鼠腸道內(nèi)較好維持;兩組移植小鼠的菌群結(jié)構(gòu)差異明顯,菌屬水平主成分分析(PCA)圖和層級(jí)聚類分析圖中兩組均能獨(dú)立分開。組間差異物種分析結(jié)果顯示:I-CMPA組與M-CMPA組中雙歧桿菌屬的比例均高于其對(duì)照組;(2)用嬰兒配方奶粉喂養(yǎng)后,M-CMPA組出現(xiàn)肛門紅腫、糞便變稀等表現(xiàn),而M-NC組沒有出現(xiàn)類似的表現(xiàn);(3)HE染色可見M-CMPA組較M-NC組出現(xiàn)明顯的空腸絨毛上皮細(xì)胞局灶性脫落,絨毛連續(xù)性破壞和炎癥細(xì)胞浸潤(rùn);(4)流式細(xì)胞分析結(jié)果顯示,M-CMPA組脾臟CD4+CD25+Foxp3+調(diào)節(jié)性T細(xì)胞比例較M-NC組降低。結(jié)論:(1)HFA小鼠能較好地模擬CMPA嬰兒的腸道微生物區(qū)系,該HFA模型為糞便菌群與CMPA的關(guān)系及機(jī)制研究提供了新的選擇;(2)糞便菌群的差異使移植小鼠對(duì)牛奶蛋白的反應(yīng)不同,糞便菌群的變化可能是CMPA發(fā)生的始動(dòng)因素,食物過敏的敏感性可通過糞菌移植的方式在不同種屬的宿主間水平傳遞;(3)雙歧桿菌屬中的某些特定菌種可能參與CMPA發(fā)生。
[Abstract]:Objective: to establish a humanized mouse model of milk protein allergic infant and normal infant fecal microflora, and to observe the reaction of the transplanted mice to milk protein.To explore the role of fecal microflora in the development of milk protein allergy.Methods: the feces of infants with gastrointestinal symptoms were collected and collected from the digestive department of Chongqing Medical University affiliated Children's Hospital, which were diagnosed as milk protein allergyra (CMPA) and were collected from the pediatric care department of Chongqing Medical University for physical examination at the same time.Normal infant feces with no gastrointestinal symptoms and no allergic diseases were treated with I-NC group, which were fed with infantile formula for .14 days after inoculation with anaphylactic mice (group M CMPA) and normal group (group M NC).The colonization and changes of fecal flora were detected by Illumina sequencing technique. The clinical manifestations of transplanted mice were observed. The pathological changes of jejunum tissue were observed by HE staining. The percentage of CD4 CD25 Foxp3 regulatory T cells in spleen cells was detected by flow cytometry.Results the sequence analysis of Illumina showed that the dominant bacteria in donor feces were different, and the richness and diversity of bacteria in I-CMPA group were lower than those in I-NC group.After 14 days of fecal bacteria transplantation, the important components of I-CMPA group could be maintained in the intestinal tract of aseptic mice, and there were significant differences between the two groups in the microflora structure of the transplanted mice, and the two groups could be separated independently from each other by PCAG and hierarchical cluster analysis.The results of species analysis showed that the proportion of Bifidobacterium in the two groups was higher than that in the control group (P < 0.05). The anus redness and fecal thinning were observed in the M-CMPA group after infantile formula was fed, and the rate of bifidobacterium in the two groups was higher than that in the control group (P < 0.05).However, there was no similar appearance in M-NC group. It was found that the focal exfoliation of chorionic villi in M-CMPA group was more obvious than that in M-NC group.The results of flow cytometry showed that the percentage of CD4 CD25 Foxp3 regulatory T cells in spleen of M-CMPA group was lower than that of M-NC group.Conclusion the microflora of the intestinal tract of CMPA infants can be well simulated by the CMPA mice. The HFA model provides a new choice for the study of the relationship and mechanism between fecal microflora and CMPA) the difference of faecal microflora makes the reaction to milk protein of transplanted mice different.The change of faecal flora may be the initiator of CMPA, and the sensitivity of food allergy can be transferred between hosts of different species by fecal bacteria transplantation. Some specific strains of Bifidobacterium may be involved in the occurrence of CMPA.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R725.9

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本文編號(hào)：1726082