粪菌移植抑制NF-κB/NLRP3改善大鼠脓毒症相关脑病

Fecal Microbiota Transplantation Inhibits NF-κB/NLRP3 Signaling to Ameliorate Sepsis-AssociatedEncephalopathy in Rats

目的探索粪菌移植(fecal microbiota transplantation,FMT)通过调节肠道菌群对SAE大鼠的神经保护作用。方法30只SD大鼠分为假手术、SAE、SAE+FMT、SAE+FMT+NF-κB激动剂,SAE+FMT+NLRP3激动剂组。通过16S rRNA测序、神经行为学评分、水迷宫测试、尼氏染色、定量逆转录聚合酶链反应和蛋白质免疫印迹试验等分析大鼠肠道菌群,神经功能及炎症反应改变。采用SPSS软件对组间多样本行单因素方差分析,进一步两两比较采用Tukey检验。结果①与假手术组相比,SAE大鼠α多样性降低(P<0.01),而FMT治疗后的SAE大鼠的α多样性升高(P<0.05),SAE组有益菌Bacteroidete,Clostridiales相比假手术组减少,FMT后增加。②与假手术组相比,SAE大鼠mNSS降低(P<0.01),学习记忆能力降低(P<0.01),海马CA1区神经元数量减少(P<0.01),而FMT治疗后的SAE大鼠的mNSS评分提高(P<0.01),学习记忆能力增加(P<0.05),神经元数量增加(P<0.05)。③与假手术组相比,SAE组大鼠肝肾功能指标、炎症相关因子、血脑屏障蛋白、NLRP3通路蛋白、NF-κB通路蛋白表达增加(P<0.05),FMT降低以上指标(P<0.05),④NF-κB和NLRP3激动剂干预后抵消了FMT的作用(P<0.05)。结论FMT通过调节肠道菌群并抑制脑内NF-κB/NLRP3信号通路。这为SAE的治疗提供了新的见解,同时强调了在临床治疗中考虑肠道微生物的重要性。

Objective To explore the effects of fecal microbiota transplantation(FMT)on SAE in rats through the modulation of the gut microbiome.Methods Total of 30 Sprague-Dawley rats were divided(random number)into sham surgery,SAE,SAE+FMT,SAE+FMT+NF-κB agonist,and SAE+FMT+NLRP3 agonist groups.The gut microbiome,neurological function,and infl ammatory responses in rats were analyzed using 16S rRNA sequencing,neurological behavioral scoring,water maze testing,Nissl staining, quantitative reverse transcription polymerase chain reaction, and western blot assays. Univariateanalysis of variance for multiple samples among groups was conducted using SPSS software, with furtherpairwise comparisons using Tukey's test. Results (1) Compared with the sham surgery group, a reduction inα-diversity was observed in the SAE rats (P<0.01), whereas an increase in α-diversity was noted in the SAErats after FMT treatment (P<0.05). A decrease in beneficial bacteria such as Bacteroidete and Clostridialeswas seen in the SAE group compared to the sham group, which increased after FMT. (2) A decrease in mNSS,learning and memory abilities, and the number of neurons in the hippocampal CA1 region was noted in SAErats compared with the sham group (P<0.01), whereas an improvement in mNSS scores, learning and memoryabilities, and neuron count was observed in SAE rats treated with FMT (P<0.05). (3) Compared with the shamgroup, increased liver and kidney function indicators, infl ammatory factors, blood-brain barrier proteins, NLRP3pathway proteins, and NF-κB pathway proteins were observed in the SAE group (P<0.05), which were reducedby FMT (P<0.05). (4) The effects of FMT were negated after the intervention with NF-κB and NLRP3 agonists(P<0.05). Conclusions FMT regulate the gut microbiome and inhibit the NF-κB/NLRP3 signaling pathwayin the brain. This provides new insights into the treatment of SAE, emphasizing the importance of consideringthe gut microbiota in clinical therapy.