M1巨噬细胞的激活通过HIF-1α-HK2信号通路调节糖酵解，从而促进糖尿病肾病。

Activation of M1 macrophages promotes diabetic kidney disease by modulating glycolysis via HIF-1α-HK2 signaling pathway.

作者信息

Pei Xiaoyan, Lu Lijuan, Huang Ziqiong, Wei Yu, Li Yu, Wang Qiong, Yang Yanli, Zhuang Langen, Jin Guoxi

机构信息

Department of Endocrinology, the First Affiliated Hospital of Bengbu Medical University, Bengbu, 233004, Anhui, P.R. China.

出版信息

Diabetol Metab Syndr. 2025 Aug 29;17(1):362. doi: 10.1186/s13098-025-01894-3.

DOI:10.1186/s13098-025-01894-3

PMID:40883798

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12395828/

Abstract

OBJECTIVE

Macrophages and glycolysis play a pivotal role in diabetic kidney disease (DKD). However, the regulation of macrophage glycolysis and its mechanism are still unclear in DKD. Thus, this research intends to investigate the regulatory mechanism of macrophage glycolysis during DKD.

METHODS

Macrophage polarization in patients with DKD and in mice was assessed using RT-qPCR and immunofluorescence. DKD was induced in the mice through injection of streptozotocin, and a high-fat diet was administered. Hematoxylin and eosin staining and Masson's trichrome staining were employed to identify pathological alterations and visualize renal collagen fibers in the model mice. Glycolytic metabolite levels were quantified using a commercially available kit. Protein levels of HK2, PFK1, LDH, PK1, and GLUT1 were analyzed via western blotting. Hkb-20 cells exposed to high glucose were utilized as an in vitro experimental model. THP-1 cells were co-cultivated with Hkb-20 cells. The impact of macrophage polarization on the functionality of Hkb-20 cells was assessed through CCK8 assay, flow cytometry, and wound healing assay. Levels of IL-6, IL-1β, and TNF-α were evaluated using ELISA kits. Immunofluorescence staining was employed to examine the co-localization of hypoxia-inducible factor-1 (HIF-1α) and HK2.

RESULTS

We found that M1 macrophage polarization and enhanced glycolysis activity emerged in DKD patients and mice. Furthermore, we demonstrated that M1 macrophage polarization promotes glycolysis activity in macrophage and model mice. Moreover, we found that HIF-1α and HK2 were obviously enhanced in the serum of DKD cases and mice. Mechanically, we revealed that HIF-1α participates in M1 macrophage polarization and glycolysis activity in vitro by activating HK2 transcription.

CONCLUSION

We demonstrated that M1 macrophage polarization mediated glycolysis contributed to DKD via regulating HIF-1α-HK2 signaling pathway.

摘要

目的

巨噬细胞和糖酵解在糖尿病肾病（DKD）中起关键作用。然而，DKD中巨噬细胞糖酵解的调节及其机制仍不清楚。因此，本研究旨在探讨DKD期间巨噬细胞糖酵解的调节机制。

方法

采用RT-qPCR和免疫荧光法评估DKD患者和小鼠的巨噬细胞极化。通过注射链脲佐菌素诱导小鼠发生DKD，并给予高脂饮食。采用苏木精-伊红染色和Masson三色染色来识别模型小鼠的病理改变并观察肾胶原纤维。使用市售试剂盒定量糖酵解代谢物水平。通过蛋白质印迹分析HK2、PFK1、LDH、PK1和GLUT1的蛋白水平。将暴露于高糖环境的Hkb-20细胞用作体外实验模型。将THP-1细胞与Hkb-20细胞共培养。通过CCK8测定、流式细胞术和伤口愈合试验评估巨噬细胞极化对Hkb-20细胞功能的影响。使用ELISA试剂盒评估IL-6、IL-1β和TNF-α的水平。采用免疫荧光染色检查缺氧诱导因子-1（HIF-1α）和HK2的共定位。