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Published on: June 23, 2013
Polygonatum Polysaccharide Regulates Macrophage Polarization and Improves LPS-Induced Acute Lung Injury through
Weizheng Zhou1, Jiang Hong1, Tao Liu1
1Department of Thoracic Surgery, Changhai Hospital, Shanghai 200433, China.
Objective:
To investigate the effects of polygonatum sibiricum polysaccharides (PSPs) on the polarization of macrophages to M1 and M2 phenotypes and their potential mechanism.
Methods:
PSPs samples were prepared through water extraction and alcohol precipitation assay. The properties of PSPs were identified and analyzed by high-performance liquid chromatography, FT-IR, and NMR assay. Then, the effects of PSPs on mouse macrophage RAW264.7 viability were measured by CCK-8 assay. The cells were randomly divided into the control group, PSPs group, LPS group, and LPS + PSPs group. M1 phenotype polarization of RAW264.7 cells was induced by LPS treatment. The effects of various treatments on expression of M2 phenotype CD206, activation of TLR4-MAPK/NF-κB signal pathway, and translocation of NF-κB into the nucleus were determined by ELISA, western blot, and immunofluorescence assay, respectively. TLR4 inhibitor, TAK-242, and MAPK inhibitor, BIRB 796, were used to verify the effects of PSPs on the TLR4-MAPK/NF-κB pathway. The mice model of acute lung injury (ALI) was established and randomly divided into control group, PSPs group, LPS group, and LPS + PSPs group. Bronchoalveolar lavage fluid (BALF) and lung tissue were collected to measure protein, inflammatory cells, neutrophil and macrophage cells number, and the levels of IL-6 and TNF-α in BALF. Flow cytometry and western blot assay measured the phenotypic changes of macrophages and the activation of the TLR4-MAPK/NF-κB signaling pathway.
Results:
The concentrations of PSPs lower than 100 μg/mL showed no toxicity to RAW264.7 cells. PSPs treatment could significantly reverse the reduction of CD206 protein expression (P < 0.05) and the increase of the expression of inflammatory factor TNF-α, IL-1β, and IL-6 (all P < 0.05), TLR4-MAPK/NF-κB signaling pathway activation (all P < 0.05), and NF-κB translocation into the nucleus induced by LPS. The effect of inhibitors TAK-242 and BIRB 796 was consistent with that of PSPs. In the mice model of ALI, PSPs treatment could reduce the total protein levels of BALF and the number of inflammatory cells level, reverse the number changes of neutrophils and macrophages, and downregulate the proinflammatory factors IL-6 and TNF-α caused by LPS (all P < 0.05). In addition, PSPs treatment could also significantly reverse the increase in the number of iNOS expressing macrophages in alveolar lavage fluid induced by LPS (P < 0.05). In contrast, CD206-expressed cells decreased (P < 0.05). PSPs could also reverse LPS-induced TLR4-MAPK/NF-κB signal pathway protein activation (all P < 0.05).
Conclusion:
PSPs could suppress TLR4-MAPK/NF-κB activation induced by LPS, inhibit M1 phenotypic polarization of macrophages, and promote M2 phenotypic polarization, thus playing an anti-inflammatory role.
Insights
Polygonatum sibiricum polysaccharides (PSPs) reduce inflammation by suppressing the TLR4-MAPK/NF-κB pathway, inhibiting M1 macrophage polarization, and promoting M2 polarization. This study highlights PSPs
Area of Science:
- Immunology
- Pharmacology
- Natural Products Chemistry
Background:
- Macrophages play a critical role in immune responses, with polarization towards M1 or M2 phenotypes influencing inflammation.
- Polygonatum sibiricum polysaccharides (PSPs) are bioactive compounds with potential immunomodulatory effects.
- Understanding the impact of PSPs on macrophage polarization is crucial for developing novel anti-inflammatory therapies.
Purpose of the Study:
- To investigate the effects of Polygonatum sibiricum polysaccharides (PSPs) on macrophage polarization to M1 and M2 phenotypes.
- To elucidate the underlying mechanisms, particularly the involvement of the TLR4-MAPK/NF-κB signaling pathway.
- To evaluate the therapeutic potential of PSPs in an acute lung injury (ALI) mouse model.
Main Methods:
- PSPs were extracted and characterized using chromatography and spectroscopy.
- RAW264.7 macrophages were treated with PSPs and lipopolysaccharide (LPS) to induce M1 polarization.
- Macrophage phenotype, inflammatory markers, and signaling pathway activation (TLR4-MAPK/NF-κB) were assessed using ELISA, Western blot, and immunofluorescence.
- An LPS-induced ALI mouse model was used to evaluate PSPs' in vivo efficacy.
Main Results:
- PSPs (below 100 μg/mL) exhibited no toxicity to RAW264.7 cells.
- PSPs reversed LPS-induced M1 polarization markers and inflammatory cytokine production (TNF-α, IL-1β, IL-6).
- PSPs suppressed TLR4-MAPK/NF-κB pathway activation and NF-κB nuclear translocation, similar to specific inhibitors.
- In vivo, PSPs reduced inflammatory cell infiltration and pro-inflammatory mediators in ALI mice, while promoting M2 markers.
Conclusions:
- PSPs effectively suppress TLR4-MAPK/NF-κB activation induced by LPS.
- PSPs inhibit M1 macrophage polarization and promote M2 polarization, demonstrating anti-inflammatory properties.
- These findings suggest PSPs hold promise as a therapeutic agent for inflammatory conditions.
