METHODS: In this study, a controlled cortical impact (CCI) model was used to induce TBI in adult male C57BL/6J mice. Subsequently, P. copri was transplanted by intragastric gavage for 7 consecutive days. To investigate the effect of the GUO-PI3K/Akt pathway in P. copri transplantation therapy, guanosine (GUO) was administered 2 h after TBI for 7 consecutive days, and PI3K inhibitor (LY294002) was administered 30 min before TBI. Various techniques were used to assess the effects of these interventions, including quantitative PCR, neurological behavior tests, metabolite analysis, ELISA, Western blot analysis, immunofluorescence, Evans blue assays, transmission electron microscopy, FITC-dextran permeability assay, gastrointestinal transit assessment, and 16 S rDNA sequencing.
RESULTS: P. copri abundance was significantly reduced after TBI. P. copri transplantation alleviated motor and cognitive deficits tested by the NSS, Morris\'s water maze and open field test. P. copri transplantation attenuated oxidative stress and blood-brain barrier damage and reduced neuronal apoptosis after TBI. In addition, P. copri transplantation resulted in the reshaping of the intestinal flora, improved gastrointestinal motility and intestinal permeability. Metabolomics and ELISA analysis revealed a significant increase in GUO levels in feces, serum and injured brain after P. copri transplantation. Furthermore, the expression of p-PI3K and p-Akt was found to be increased after P. copri transplantation and GUO treatment. Notably, PI3K inhibitor LY294002 treatment attenuated the observed improvements.
CONCLUSIONS: We demonstrate for the first time that P. copri transplantation can improve GI functions and alter gut microbiota dysbiosis after TBI. Additionally, P. copri transplantation can ameliorate neurological deficits, possibly via the GUO-PI3K/Akt signaling pathway after TBI.
方法:在本研究中,在成年雄性C57BL/6J小鼠中使用受控皮质冲击(CCI)模型来诱导TBI。随后,通过灌胃连续7天进行P.copri移植。探讨GUO-PI3K/Akt通路在同色假单胞菌移植治疗中的作用,鸟苷(GUO)在TBI后2小时连续7天,和PI3K抑制剂(LY294002)在TBI前30分钟给药。使用各种技术来评估这些干预措施的效果,包括定量PCR,神经行为测试,代谢物分析,ELISA,蛋白质印迹分析,免疫荧光,伊文思蓝检测,透射电子显微镜,FITC-葡聚糖通透性测定,胃肠转运评估,和16SrDNA测序。
结果:P.TBI后,Copri丰度显着降低。P.combri移植减轻了NSS测试的运动和认知缺陷,莫里斯的水迷宫和野外试验。P.combri移植减轻氧化应激和血脑屏障损伤,减少TBI后神经元凋亡。此外,P.combri移植导致肠道菌群的重塑,改善胃肠蠕动和肠通透性。代谢组学和ELISA分析显示粪便中GUO水平显着增加,P.combri移植后血清和脑损伤。此外,发现p-PI3K和p-Akt的表达在P.combri移植和GUO治疗后增加。值得注意的是,PI3K抑制剂LY294002治疗减弱了观察到的改善。
结论:我们首次证明P.copiri移植可以改善TBI后的胃肠道功能并改变肠道菌群失调。此外,P.combri移植可以改善神经功能缺损,TBI后可能通过GUO-PI3K/Akt信号通路。