PDF(Pubmed)
Abstract：
Amyotrophic lateral sclerosis (ALS) is a neuromuscular disease. The ALS mice expressing human mutant of transactive response DNA binding protein of 43 kDa (hmTDP43) showed intestinal dysfunction before neuromuscular symptoms. We hypothesize that restoring the intestinal and microbial homeostasis with a bacterial metabolite or probiotics delays the ALS disease onset. We investigate the pathophysiological changes in the intestine and neurons, intestinal and blood-brain barriers, and inflammation during the ALS progression. We then cultured enteric glial cells (EGCs) isolated from TDP43 mice for mechanistic studies. TDP43 mice had significantly decreased intestinal mobility, increased permeability, and weakened muscle, compared with the age-matched wild-type mice. We observed increased hmTDP43 and Glial fibrillary acidic protein (GFAP), and decreased expression of α-smooth muscle actin (α-SMA), tight junction proteins (ZO-1 and Claudin-5) in the colon, spinal cord, and brain in TDP43 mice. TDP43 mice had reduced Butyryl-coenzyme A CoA transferase, decreased butyrate-producing bacteria Butyrivibrio fibrisolvens, and increased Bacteroides fragilis, compared to the WT mice. Serum inflammation cytokines (IL-6, IL-17, and IFN-γ) and LPS were elevated in TDP43 mice. EGCs from TDP43 mice showed aggregation of hmTDP43 associated with increased GFAP and ionized calcium-binding adaptor molecule (IBA1, a microglia marker). TDP43 mice treated with butyrate or probiotic VSL#3 had significantly increased rotarod time, increased intestinal mobility and decreased permeability, compared to the untreated group. Butyrate or probiotics treatment decreased the expression of GFAP, TDP43, and increased α-SMA, ZO-1, and Claudin-5 in the colon, spinal cord, and brain. Also, butyrate or probiotics treatment enhanced the Butyryl-coenzyme A CoA transferase, Butyrivibrio fibrisolvens, and reduced inflammatory cytokines in TDP43 mice. The TDP43 EGCs treated with butyrate or probiotics showed reduced GFAP, IBA1, and TDP43 aggregation. Restoring the intestinal and microbial homeostasis by beneficial bacteria and metabolites provide a potential therapeutic strategy to treat ALS.
摘要：
肌萎缩侧索硬化症(ALS)是一种神经肌肉疾病。表达43kDa的反应性DNA结合蛋白（hmTDP43）的人突变体的ALS小鼠在出现神经肌肉症状之前表现出肠功能障碍。我们假设用细菌代谢物或益生菌恢复肠道和微生物稳态会延迟ALS疾病的发作。我们调查肠道和神经元的病理生理变化，肠和血脑屏障，和ALS进展期间的炎症。然后,我们培养从TDP43小鼠分离的肠神经胶质细胞(EGC)用于机理研究。TDP43小鼠肠道活动能力明显下降，渗透率增加，和虚弱的肌肉，与年龄匹配的野生型小鼠相比。我们观察到hmTDP43和胶质纤维酸性蛋白（GFAP）增加，α-平滑肌肌动蛋白（α-SMA）的表达降低，结肠中的紧密连接蛋白（ZO-1和Claudin-5），脊髓,和TDP43小鼠的大脑。TDP43小鼠丁酰辅酶A辅酶A转移酶减少，减少产生丁酸的细菌Butyrivibriofibrisvens，和增加的脆弱拟杆菌，与WT小鼠相比。在TDP43小鼠中,血清炎症细胞因子(IL-6、IL-17和IFN-γ)和LPS升高。来自TDP43小鼠的EGC显示hmTDP43的聚集与增加的GFAP和离子化的钙结合衔接分子(IBA1,小胶质细胞标志物)相关。用丁酸盐或益生菌VSL#3处理的TDP43小鼠具有显著增加的旋转时间,增加肠道流动性和降低渗透性，与未治疗组相比。丁酸盐或益生菌处理可降低GFAP的表达，TDP43和增加的α-SMA，结肠中的ZO-1和Claudin-5,脊髓,和大脑。此外，丁酸盐或益生菌治疗增强了丁酰辅酶A辅酶A转移酶，Butyrivibriofibrisolvens，和减少TDP43小鼠的炎性细胞因子。用丁酸盐或益生菌处理的TDP43EGCs显示GFAP降低，IBA1和TDP43聚合。通过有益细菌和代谢物恢复肠道和微生物体内平衡提供了治疗ALS的潜在治疗策略。
