Probiotic-derived nano-particles in alcoholic liver disease

益生菌衍生的纳米颗粒治疗酒精性肝病

• 批准号: 10625858
• 负责人: WENKE FENG
• 金额: $ 20.7万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-05-15 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10625858
• 关键词: Acute; Agonist; Alcohol-Induced Disorders; Alcoholic Hepatitis; Alcoholic Liver Diseases; Alcohols; Aryl Hydrocarbon Receptor; Bacteria; Bacterial Translocation; Bilophila wadsworthia; Biological Markers; Cells; Chronic; Clinical; Development; Digestive System Disorders; Disease; Endocytosis; Environment; Epithelial Cells; Epithelium; Evaluation; Experimental Models; Functional disorder; Future; Ginger; Growth; Homeostasis; Intestines; Mediating; Mus; National Institute on Alcohol Abuse and Alcoholism; Natural regeneration; Outcome; Oxidative Stress Induction; Pathogenicity; Pathway interactions; Patients; Play; Prevention; Probiotics; Production; Receptor Signaling; Regenerating islet derived protein 3-Gamma; Role; Sampling; Signal Transduction; Testing; Therapeutic Effect; Tight Junctions; Tryptophan; Up-Regulation; alcohol abuse therapy; alcohol exposure; aryl hydrocarbon receptor ligand; clinical application; disorder prevention; dysbiosis; exosome; gut colonization; gut inflammation; gut microbes; gut microbiota; improved; interleukin-22; intestinal barrier; intestinal epithelium; islet; metabolomics; microbial; mouse model; nanoparticle; nutrition; organ injury; pathogenic bacteria; prebiotics; prevent; probiotic therapy; programs; protective effect; transcytosis

Probiotics have been used to prevent/treat alcoholic liver disease (ALD). Live probiotics need to colonize the gut to exert their function. Unfortunately, underlying disease states provide an unfavorable environment for probiotic bacterial gut colonization, which diminishes probiotics’ function. In the last few years, we showed that LGG culture supernatant (LGGs, without live bacteria) was effective in the prevention of ALD in experimental models of acute and chronic alcohol exposure in mice. However, how LGG supernatant exerts its therapeutic effects is not fully understood. Exosomes are nanoparticles (NPs) derived from cell endocytosis which act as transmitters between cells. Recent studies show that bacteria, both Gram-negative and Gram-positive, produce NPs. The NPs derived from “bad” bacteria have been demonstrated to be pathogenic. However, “good” bacteria-, probiotics-derived NPs have not been studied. Our preliminary study showed that administration of LGG-derived exosome-like NPs (LDNPs) effectively reversed ALD in the binge-on-chronic alcohol exposure mouse model, suggesting that probiotic LGGs may exert its function through LDNPs in ALD. Administration of LDNPs markedly increased intestinal AhR activity, IL-22, and regenerating islet-derived 3 (Reg3) β and γ expression, which play a key role in maintaining gut microbiota homeostasis and preventing bacterial intestinal transcytosis. In addition, LDNPs administration significantly increased intestinal epithelial cell (IEC) tight junctions and decreased circulating LPS concentration associated with upregulation of intestinal Nrf2 signaling, which is known for protecting intestinal barrier junctions against oxidative stress-induced damage by alcohol. Metabolomic analysis revealed that LDNPs contain high levels of microbial metabolites of tryptophan, which are AhR ligands, indicating LDNPs may activate intestinal AhR signaling. Furthermore, we demonstrated that ginger exosome-like nanoparticles (GDNPs) are preferentially taken up by LGG, suggesting that GDNP may serve as a prebiotic to enhance the effects of LGG. These preliminary studies provide the groundwork for our central hypothesis that, by activating intestinal AhR-Nrf2 signaling, LDNPs increase intestinal expression of Il-22, Reg3 and tight junctions, and modulate gut microbiota homeostasis and enhance intestinal barrier function, leading to the suppression of ALD. We will test our hypothesis in following four specific aims: (1) Determine the role of bacteria- derived NPs in ALD; (2) Define the mechanisms of the beneficial effects of LDNPs in ALD; (3) Determine whether GDNPs treatment enhances LDNP production and AhR agonist enrichment that lead to improved effects of LDNPs against ALD; and (4) Evaluate the intestinal AhR-Nrf2 signaling in alcoholic hepatitis patients treated with LGG. Completion of this study is expected to significantly impact the development of LGG-based probiotic therapeutics in the treatment of alcohol-associated liver diseases.

益生菌已用于预防/治疗酒精性肝病（ALD）。实时益生菌需要定居肠道 执行其功能。不幸的是，潜在的疾病状态为益生菌提供了一个不利的环境 细菌肠道定殖，可降低益生菌的功能。在过去的几年中，我们证明了LGG 在实验模型中，培养上清液（LGGS，无活细胞）在预防ALD方面有效 小鼠的急性和慢性酒精暴露。但是，LGG上清液如何执行其治疗作用是 不完全理解。外泌体是源自细胞内吞作用的纳米颗粒（NP），其充当发射机 在细胞之间。最近的研究表明，革兰氏阴性和革兰氏阳性的细菌产生了NP。 源自“不良”细菌的NP已被证明是致病性的。但是，“好”细菌 - 尚未研究益生菌衍生的NP。我们的初步研究表明，LGG衍生的给药 外泌体样的NP（LDNP）有效地逆转了气味饮酒小鼠模型中的ALD， 表明益生菌LGG可以通过ALD中的LDNP执行其功能。明显管理LDNP 肠道AHR活性增加，IL-22和再生胰岛衍生的3（reg3）β和γ表达，它们的发挥作用 维持肠道菌群稳态和预防细菌肠道胞症的关键作用。此外， LDNPS给药可显着增加肠上皮细胞（IEC）紧密连接并改善 循环LPS浓度与肠NRF2信号的上调有关，该信号传导闻名 保护肠道屏障连接处免受酒精氧化应激引起的损害。代谢组分析 揭示了LDNP含有高水平的色氨酸的微生物代谢产物，即AHR配体，表明 LDNP可能会激活肠道AHR信号传导。此外，我们证明了姜外泌体状 纳米颗粒（GDNP）优先用LGG吸收，这表明GDNP可以用作益生元 增强LGG的影响。这些初步研究为我们的中心假设提供了基础，即 通过激活肠道AHR-NRF2信号传导，LDNP会增加IL-22，REG3和TICT的肠道表达 连接，调节肠道菌群稳态并增强肠屏障功能，导致 抑制ALD。我们将在以下四个特定目标中检验我们的假设：（1）确定细菌的作用 ALD派生的NP； （2）定义LDNP在ALD中的有益作用的机制； （3）确定是否 GDNPS治疗增强LDNP产生和AHR激动剂富集，从而改善了 针对ALD的LDNP； （4）评估接受治疗的酒精性肝炎患者的肠道AHR-NRF2信号传导 lgg。预计这项研究的完成将显着影响基于LGG的益生菌的发展 治疗酒精相关肝病的治疗剂。