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芪黄汤通过lncRNA MALAT1对miR-146a的负调控来调节巨噬细胞M1极化,从而抑制细菌移位。

Qihuang decoction inhibits bacterial translocation by modulating macrophage M1 polarisation via negative regulation of miR-146a by lncRNA MALAT1.

作者信息

Zhou Fuhai, Li Ziyi, Lu Erguang, Peng Hui, Yu Qingsheng

机构信息

Department of General Surgery, The First Affiliated Hospital of Anhui University of Traditional Chinese Medicine, Hefei, China.

Department of General Surgery, The First Affiliated Hospital of Anhui University of Traditional Chinese Medicine, Hefei, China.

出版信息

Microb Pathog. 2025 Oct;207:107895. doi: 10.1016/j.micpath.2025.107895. Epub 2025 Jul 12.

Abstract

OBJECTIVE

This study aimed to investigate whether Qihuang decoction prevents bacterial lipopolysaccharide (LPS) translocation by modulating macrophage M1 polarisation.

METHODS

A rat model of sleeve gastrectomy was established. Haematoxylin-eosin staining and electron microscopy were used to observe changes in small intestinal structures. Polymerase chain reaction (PCR), Western blotting and enzyme-linked immunosorbent assay (ELISA) were employed to detect expression levels of proteins and genes related to the TLR4/NF-κB pathway. Detection of phosphorylated NF-κB (p-NF-κB) was included to directly evaluate pathway activation. The expression levels of lncMALAT1 and miR-146a were also assessed by PCR. In addition, fluorescein isothiocyanate-labelled dextran (FD4), ELISA and a D-lactate assay kit were used to measure FD4 clearance, LPS levels and lactate concentrations. Macrophage polarisation was evaluated by flow cytometry and immunohistochemistry for iNOS, and CD206 was performed to visualise M1/M2 macrophage distribution in intestinal tissues.

RESULTS

The small intestine remained structurally intact, and inflammatory cell infiltration was reduced in the Qihuang decoction treatment group compared with the model group. There was a substantial decrease in the proportion of M1-polarised macrophages, as confirmed by both flow cytometry and immunohistochemistry. Qihuang decoction treatment resulted in substantial downregulation of lncRNA MALAT1, increased expression of miR-146a and reduced expression of TLR4/NF-κB pathway-related genes and proteins, including p-NF-κB. Lactate concentration, FD4 clearance and LPS levels were also reduced.

CONCLUSION

These findings suggest that Qihuang decoction negatively regulates miR-146a via lncRNA MALAT1, thereby reducing macrophage M1 polarisation induced by the TLR4/NF-κB pathway to inhibit bacterial translocation.

摘要

目的

本研究旨在探讨芪黄汤是否通过调节巨噬细胞M1极化来预防细菌脂多糖(LPS)移位。

方法

建立大鼠袖状胃切除术模型。采用苏木精-伊红染色和电子显微镜观察小肠结构变化。运用聚合酶链反应(PCR)、蛋白质免疫印迹法和酶联免疫吸附测定(ELISA)检测与Toll样受体4/核因子κB(TLR4/NF-κB)通路相关的蛋白质和基因表达水平。检测磷酸化核因子κB(p-NF-κB)以直接评估通路激活情况。还通过PCR评估长链非编码RNA MALAT1和微小RNA-146a(miR-146a)的表达水平。此外,使用异硫氰酸荧光素标记的葡聚糖(FD4)、ELISA和D-乳酸检测试剂盒测量FD4清除率、LPS水平和乳酸浓度。通过流式细胞术和免疫组织化学检测诱导型一氧化氮合酶(iNOS)评估巨噬细胞极化,并进行CD206检测以观察肠道组织中M1/M2巨噬细胞分布情况。

结果

与模型组相比,芪黄汤治疗组小肠结构保持完整,炎症细胞浸润减少。流式细胞术和免疫组织化学均证实,M1极化巨噬细胞比例显著降低。芪黄汤治疗导致长链非编码RNA MALAT1显著下调、miR-146a表达增加,以及TLR4/NF-κB通路相关基因和蛋白质(包括p-NF-κB)表达减少。乳酸浓度、FD4清除率和LPS水平也降低。

结论

这些研究结果表明,芪黄汤通过长链非编码RNA MALAT1对miR-146a进行负调控,从而减少TLR4/NF-κB通路诱导的巨噬细胞M1极化,以抑制细菌移位。

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