治疗大肠杆菌性犊牛腹泻病的蒙兽药及其有效成分的调理肠道菌群、抗炎免疫机理研究

Bacterial resistance to antibiotics and animal food antibiotic residues, become a major problem that affects the human and animal health and food safety. To this end, use herb medicines regulating the body's physiological function, improve the anti-inflammatory increasing-immune to become one of the trends. E. coli diarrhea is a main reason of causing calf death and growth retardation. Mongolian medicine babua 7 in clinical cure diarrhea is more than 95%, has antibacterial anti-inflammatory increasing-immune function. For the study of Mongolian veterinary medicine treatment diarrhea in regulating-bacterium of intestine anti-inflammatory immune mechanism caused by bovine coming wild E. coli, physiology, pharmacology, molecular biology methods, ELISA and flow cytometry technology be used, to research the role of blood immune index in its pathogenesis, the influence mechanism causing diarrhea E coli in intestinal mucosal mechanical barrier and immune barrier, including the Payer 's knot, in cell and molecular level; to research Mongolian veterinary medicine and its effective components on regulating intestinal bacterial, treatment diarrhea mechanism by influencing of anti-inflammatory and immune function, the intestinal mucosal mechanical and immune barrier mechanism, and to research cell signal transduction mechanism of its anti-inflammatory increasing-immune, screening mainly activated gene, to explore anti-inflammatory immune mechanism of Mongolian veterinary medicine and its effective ingredients on the level of gene and protein expression . To provides the theory basis for prevent and control harmful calf diarrhea disease, high-yielding healthy cows with Mongolian instead of antibiotics. It is the original research on the early basis stage.

细菌对抗生素产生耐药性和动物性食品抗生素残留，成为影响人畜健康和食品安全的重大难题。为此，用草药等调控机体生理机能、提高抗炎免疫力成为趋势之一。大肠杆菌腹泻是犊牛死亡和发育阻滞的主因。蒙药巴布7临床治愈腹泻达95%以上，具有抑菌抗炎免疫作用。为研究蒙兽药在牛源野生大肠杆菌性腹泻中调理肠道菌群及抗炎免疫作用机理，采用生理、药理、分子生物学方法、ELISA、流式细胞术等，在细胞分子层面研究血液免疫指标在其发病机制中的作用、致腹泻病大肠杆菌对肠黏膜机械和免疫屏障包括Payer's结的影响机理；研究蒙兽药及其有效成分调理肠道菌群、治疗腹泻对机体抗炎免疫机能、肠黏膜机械和免疫屏障的影响机制，并通过抗炎免疫细胞信号转导机制研究，筛选主要激活的上下调基因，从基因和蛋白表达水平探索蒙兽药有效成份的抗炎免疫机理。为防控危害犊牛健康的腹泻病、培育高产健康牛和蒙药替代抗生素提供理论依据。是前期研究基础上的创新。

为了解治疗犊牛腹泻病的蒙药及有效成分复方的作用机理，正交设计筛选出最佳疗效抑菌抗炎免疫成分复方；研究其调理肠道菌群、提高机体抗炎免疫机能、肠黏膜屏障功能，治疗腹泻的影响机制；通过抗炎免疫细胞信号转导机制研究，筛选主要激活的上下调基因，探索蒙兽药有效成份的抗炎免疫机理。结果：正交设计筛选出蒙兽药复方及其有效成分复方，E. coli O1对其高敏抑菌。经体外抑菌推测是通过改变该菌氧化还原酶活性，改变其毒力、感染力下降；转录分析（DEGs），通路富集（KEGG）分析与GO富集分析注释了86条途径，其中5条显著富集的KEGG途径；而硫代谢途径和不同环境中微生物代谢信号通路中富集大量上调基因，极可能是其降低E.coli O1致病的重要原因。口服给药，通过自身抑菌特性直接抑制致病性大肠杆菌繁殖，通过增加胃肠道有益菌群丰富度形成肠黏膜生物学屏障发挥保护功能。有效修复致病性E.coliO1对小肠的病理损伤，改善小肠消化吸收功能，提高紧密连接相关蛋白表达，抑制因E.coliO1对杯状细胞损伤造成的粘液分泌降低与小肠黏膜通透性升高等现象持续维持黏液更新，从而修复受损的肠黏膜化学与机械屏障功能。通过调控PGE2、细胞间粘附分子-1、E-选择素和NO等炎症介质抑制毛细血管通透性升高、炎症局部肿胀和增生。其抗炎作用与地塞米松相近，可消除致炎因子，降低炎症对局部组织造成的损伤。显著提高小鼠及犊牛非特异和特异性免疫力，进而减少炎症细胞因子、提高抗炎因子水平而减轻炎症。通过对神经递质的调节减少肠非正常蠕动、缓解腹痛。在降低犊牛腹泻率的同时维持腹泻犊牛体重、有利增重，为后期保育工作打好基础，治疗犊牛腹泻病疗效与环丙沙星相近。蒙兽药及其有效成分复方通过调理肠道菌群和神经-内分泌活动提高抗炎免疫机能、保护肠黏膜化学、机械和免疫屏障等功能，提高犊牛免疫与生产能力。蒙兽药及有效成分复方急、亚慢性毒理学在安全使用剂量范围无毒。该研究为蒙药及其成分复方替代抗生素，治疗犊牛腹泻病提供了依据。