Related Experiment Video
Updated: Jul 27, 2026
Investigation of Macrophage Polarization Using Bone Marrow Derived Macrophages
Published on: June 23, 2013
Effect of regulating macrophage polarization phenotype on intervertebral disc degeneration
Xuefeng Hou1, Yucheng Shen1, Minli Sun2
1Department of Orthopedics, Binhai County People's Hospital, Jiangsu Province, China.
Background:
Macrophages are the only inflammatory cells that can penetrate the closed nucleus pulposus and their polarization plays an important role in intervertebral disc degeneration (IVDD). This paper attempted to investigate the pathogenesis of IVDD by altering the polarization state of macrophages.
Methods:
Macrophage RAW264.7 cells were induced by interferonγ (IFN-γ) and lipopolysaccharide (LPS). The polarization of RAW264.7 cells was estimated by western blot and immunofluorescence. The expressions of inflammatory factors were detected by ELISA. Subsequently, RAW264.7 cells were treated with different concentrations of minocycline (Mino) and sinomenine (Sino), followed by the assessment of cell viability with cell counting kit-8 kit. Then, RAW264.7 cell culture medium was collected for the culture of human nucleus pulposus cells (NPCs). Toluidine blue staining and type II collagen staining were applied to assay the level of type II collagen. The cell apoptosis, oxidative stress, and nitric oxide (NO) level were appraised by TUNEL, oxidative stress kits and NO kit, respectively. Western blot was employed to test the levels of apoptosis- and oxidative stress-related proteins.
Results:
IFN-γ and LPS could induce M1 polarization of RAW264.7 cells. Mino and Sino could reduce the polarization of RAW264.7 cells toward M1. M1-polarized medium inhibited LPS-induced activity, inflammation, and damage of NPCs, which were enhanced by Mino and Sino in medium.
Conclusion:
M1 polarization of macrophages promoted LPS-induced inflammation and damage of NPCs.
Insights
Macrophage polarization contributes to intervertebral disc degeneration. Minocycline and sinomenine reduced M1 polarization, mitigating inflammation and damage in nucleus pulposus cells.
Area of Science:
- Immunology
- Cell Biology
- Biochemistry
Background:
- Macrophages infiltrate the nucleus pulposus, a key factor in intervertebral disc degeneration (IVDD).
- Macrophage polarization significantly influences the pathogenesis of IVDD.
Purpose of the Study:
- To investigate the role of macrophage polarization in IVDD pathogenesis.
- To explore the potential of modulating macrophage polarization to treat IVDD.
Main Methods:
- RAW264.7 macrophage cells were polarized using interferon-gamma (IFN-γ) and lipopolysaccharide (LPS).
- Minocycline (Mino) and sinomenine (Sino) were used to modulate macrophage polarization.
- Effects on human nucleus pulposus cells (NPCs) were assessed, including viability, apoptosis, oxidative stress, and inflammation.
Main Results:
- IFN-γ and LPS induced M1 polarization in RAW264.7 cells.
- Mino and Sino treatment reduced M1 polarization.
- M1-polarized macrophage medium exacerbated LPS-induced inflammation and damage in NPCs, while Mino and Sino enhanced NPC protection.
Conclusions:
- M1 macrophage polarization promotes inflammation and damage in nucleus pulposus cells, contributing to IVDD.
- Modulating macrophage polarization with Mino and Sino shows therapeutic potential for IVDD.
More Related Videos
07:45Metabolic Characterization of Polarized M1 and M2 Bone Marrow-derived Macrophages Using Real-time Extracellular Flux Analysis
Published on: November 28, 2015
05:46A Proinflammatory, Degenerative Organ Culture Model to Simulate Early-Stage Intervertebral Disc Disease.
Published on: February 14, 2021
Related Concept Videos
01:27Degenerative Disc Disease I: Introduction
01:23Degenerative Disc Disease ll: Pathophysiology