Effect of stromal cell-derived factor 1 on macrophage polarization through PI3K/AKT1 signaling pathway

doi:10.3877/cma.j.issn.1674-1366.2024.02.003

Abstract

Abstract:

Objective

To explore the effect and mechanism of stromal cell-derived factor 1 (SDF-1) on migration and polarization of mouse RAW264.7 macrophages.

Methods

RAW264.7 macrophages in vitro were treated with SDF-1 and/or phosphotidylinsitol-3-kinase (PI3K) inhibitor LY294002. Transwell was used to detect cell migration. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of related genes in each group. The specific surface markers of M1 and M2 macrophages were detected by flow cytometry. Cytokines were determined by ELISA. The phosphorylation level of PI3K/AKT1 protein was detected by Western blot.

Results

SDF-1 could promote the migration of macrophages. The number of migrating cells in SDF-1 group increased from (90 ± 16) to (199 ± 9) (t = 12.010, P<0.001) and the expression levels of anti-inflammatory related genes IL-10 and TGF-β significantly increased during the induction of M2 polarization in macrophages. The expression of IL-10 increased from (1.015 ± 0.111) to (3.686 ± 0.268) (t = 15.960, P<0.001) . The proportion of CD206+ M2-like macrophages increased by 11.5%. The phosphorylation level of PI3K/AKT1 protein was significantly increased, and the expression of p-AKT1 increased from (1.02 ± 0.09) to (1.47 ± 0.12) (t = 5.082, P = 0.007) . which promoted the polarization of macrophages towards M2. LY294002 inhibited the phosphorylation of PI3K/AKT1 protein, and the expression of p-Akt1 decreased from (1.02 ± 0.09) to (0.41 ± 0.13) (t = 6.503, P = 0.002) . The migration ability of macrophages induced by SDF-1 was decreased from (90 ± 16) to (60 ± 11) (t = 3.133, P = 0.02) . At the same time, the expression levels of anti-inflammatory related factors IL-10 and TGF-β during the polarization of M2 macrophages were down-regulated. The expression of IL-10 decreased from (1.015 ± 0.111) to (0.608 ± 0.034) (t = 6.075, P<0.001) . The proportion of CD206+ M2-like macrophages decreased by 10.3%. Compared with the SDF-1 treatment group, the expression of anti-inflammatory related factors IL-10 and TGF-β in the polarization of M2 macrophages in the SDF-1 and LY294002 combined treatment group decreased (t_IL-10 = 14.730, P_IL-10<0.001, t_TGF-β = 31.180, P_TGF-β<0.001) , while the expression of pro-inflammatory factors IL-6 and TNF-α in the polarization of M1 macrophages did not change significantly.

Conclusion

SDF-1 could significantly promote the M2 polarization of macrophages, possibly by activating the PI3K/AKT1 signaling pathway.

Key words: Stromal cell-derived factor 1, Macrophage, Macrophage polarization, PI3K/AKT1 signaling pathway

Jingyi Di, Yujiang Chen, Xinxin Chen, Wenxia Chen. Effect of stromal cell-derived factor 1 on macrophage polarization through PI3K/AKT1 signaling pathway[J]. Chinese Journal of Stomatological Research(Electronic Edition), 2024, 18(02): 89-95.

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Figures/Tables 6

References 21

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基因	引物序列
IL-6	5′-CTTCTTGGGACTGATGCTGGTGAC-3′
	5′-TCTGTTGGGAGTGGTATCCTCTGTG-3′
TNF-α	5′-GGACTAGCCAGGAGGGAGAACAG-3′
	5′-GCCAGTGAGTGAAAGGGACAGAAC-3′
IL-10	5′-AGAGAAGCATGGCCCAGAAATCAAG-3′
	5′-CTTCACCTGCTCCACTGCCTTG-3′
TGF-β	5′-ACCGCAACAACGCCATCTATGAG-3′
	5′-GGCACTGCTTCCCGAATGTCTG-3′
GAPDH	5′-GGCAAATTCAACGGCACAGTCAAG-3′
	5′-TCGCTCCTGGAAGATGGTGATGG-3′

基因	引物序列
IL-6	5′-CTTCTTGGGACTGATGCTGGTGAC-3′
	5′-TCTGTTGGGAGTGGTATCCTCTGTG-3′
TNF-α	5′-GGACTAGCCAGGAGGGAGAACAG-3′
	5′-GCCAGTGAGTGAAAGGGACAGAAC-3′
IL-10	5′-AGAGAAGCATGGCCCAGAAATCAAG-3′
	5′-CTTCACCTGCTCCACTGCCTTG-3′
TGF-β	5′-ACCGCAACAACGCCATCTATGAG-3′
	5′-GGCACTGCTTCCCGAATGTCTG-3′
GAPDH	5′-GGCAAATTCAACGGCACAGTCAAG-3′
	5′-TCGCTCCTGGAAGATGGTGATGG-3′

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