通过肠道微生物组分析和基于 LC-MS 的代谢组学研究丁酸梭菌治疗急性胰腺炎中 AMPK/NF-κB 的调控作用。

Gut microbiota analysis and LC-MS-based metabolomics to investigate AMPK/NF-κB regulated by Clostridium butyricum in the treatment of acute pancreatitis.

机构信息

Department of Gastroenterology, The Second Affiliated Hospital, School of Medicine, South China University of Technology, Guangzhou, 510013, China.

Department of Gastroenterology, Guangzhou First People's Hospital, South China University of Technology, Guangzhou, 510180, China.

出版信息

J Transl Med. 2024 Nov 27;22(1):1072. doi: 10.1186/s12967-024-05764-w.

DOI:10.1186/s12967-024-05764-w

PMID:39604956

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11600808/

Abstract

BACKGROUND

Acute pancreatitis (AP) is an inflammatory condition with potentially life-threatening complications. This study investigates the therapeutic potential of Clostridium butyricum for modulating the inflammatory cascade through the AMPK/NF-κB signaling pathway, focusing on inflammation induced by AP. LC-MS analysis of serum samples from AP patients highlighted the regulation of lipid metabolism and inflammation, and found that metabolites involved in the inhibition of NF-κB phosphorylation and the AMPK activation pathway were downregulated. We hypothesized that pre-administration of Clostridium butyricum and its culture supernatant could mitigate AP-induced damage by modulating the AMPK/NF-κB pathway.

METHODS

Lipopolysaccharide (LPS)-induced cell inflammation models. LPS combined with CAE induced acute pancreatitis in mice. We divided mice into four groups: Con, AP, AP + C.Buty (AP with Clostridium butyricum treatment), and AP + CFS (AP with culture supernatant treatment). Analyses were performed using WB, RT-qPCR, Elisa, flow cytometry, IHC, and HE, respectively.

RESULTS

Our study shows that CFS can reduce the apoptosis of LPS-induced cellular inflammation and reduce the release of LPS-induced cytoinflammatory factors through the AMPK/NF-κB pathway in vitro. In vivo, Clostridium butyricum and its supernatant significantly reduced inflammatory markers, and corrected histopathological alterations in AP mice. Gut microbiota analysis further supported these results, showing that Clostridium butyricum and its supernatant could restore the balance of intestinal flora disrupted by AP.

CONCLUSIONS

Mechanistically, our results indicated that the therapeutic effects of Clostridium butyricum are mediated through the activation of AMPK, leading to the inhibition of the NF-κB pathway, thereby reducing the production of pro-inflammatory cytokines. Clostridium butyricum and its culture supernatant exert a protective effect against AP-induced damage by modulating the AMPK/NF-κB signaling pathway. Future studies will further elucidate the molecular mechanisms underlying the beneficial effects of Clostridium butyricum in AP and explore its clinical applicability in human subjects.

摘要

背景

急性胰腺炎（AP）是一种具有潜在生命威胁的炎症性疾病。本研究通过 AMPK/NF-κB 信号通路探讨丁酸梭菌在调节炎症级联反应方面的治疗潜力，重点关注 AP 引起的炎症。对 AP 患者血清样本进行 LC-MS 分析，发现脂质代谢和炎症受到调节，且抑制 NF-κB 磷酸化和 AMPK 激活途径的代谢物下调。我们假设丁酸梭菌及其培养上清液的预处理可以通过调节 AMPK/NF-κB 通路来减轻 AP 引起的损伤。

方法

LPS 诱导的细胞炎症模型。LPS 联合 CAE 诱导小鼠急性胰腺炎。我们将小鼠分为四组：Con、AP、AP+C.Buty（AP 加丁酸梭菌治疗）和 AP+CFS（AP 加培养上清液治疗）。分别采用 WB、RT-qPCR、Elisa、流式细胞术、免疫组化和 HE 进行分析。

结果

本研究表明，CFS 可通过 AMPK/NF-κB 通路减少 LPS 诱导的细胞炎症中的细胞凋亡并减少 LPS 诱导的细胞因子的释放。在体内，丁酸梭菌及其上清液可显著降低炎症标志物，并纠正 AP 小鼠的组织病理学改变。肠道微生物组分析进一步支持了这些结果，表明丁酸梭菌及其上清液可恢复 AP 破坏的肠道菌群平衡。

结论

从机制上讲，我们的结果表明丁酸梭菌的治疗效果是通过激活 AMPK 介导的，从而抑制 NF-κB 通路，从而减少促炎细胞因子的产生。丁酸梭菌及其培养上清液通过调节 AMPK/NF-κB 信号通路对 AP 引起的损伤发挥保护作用。未来的研究将进一步阐明丁酸梭菌在 AP 中的有益作用的分子机制，并探索其在人类受试者中的临床适用性。